Coulomb fission in multiply charged molecular clusters: Experiment and theory

NASA Astrophysics Data System (ADS)

Harris, Christopher; Baptiste, Joshua; Lindgren, Eric B.; Besley, Elena; Stace, Anthony J.

2017-04-01

A series of three multiply charged molecular clusters, (C6H6)nz+ (benzene), (CH3CNnz) + (acetonitrile), and (C4H8O)nz+ (tetrahydrofuran), where the charge z is either 3 or 4, have been studied for the purpose of identifying the patterns of behaviour close to the charge instability limit. Experiments show that on a time scale of ˜10-4 s, ions close to the limit undergo Coulomb fission where the observed pathways exhibit considerable asymmetry in the sizes of the charged fragments and are all associated with kinetic (ejection) energies of between 1.4 and 2.2 eV. Accurate kinetic energies have been determined through a computer simulation of peak profiles recorded in the experiments and the results modelled using a theory formulated to describe how charged particles of dielectric materials interact with one another [E. Bichoutskaia et al., J. Chem. Phys. 133, 024105 (2010)]. The calculated electrostatic interaction energy between separating fragments gives an accurate account for the measured kinetic energies and also supports the conclusion that +4 ions fragment into +3 and +1 products as opposed to the alternative of two +2 fragments. This close match between the theory and experiment reinforces the assumption that a significant fraction of excess charge resides on the surfaces of the fragment ions. It is proposed that the high degree of asymmetry seen in the fragmentation patterns of the multiply charged clusters is due, in part, to limits imposed by the time window during which observations are made.

Beyond the Rayleigh instability limit for multicharged finite systems: From fission to Coulomb explosion

PubMed Central

Last, Isidore; Levy, Yaakov; Jortner, Joshua

2002-01-01

We address the stability of multicharged finite systems driven by Coulomb forces beyond the Rayleigh instability limit. Our exploration of the nuclear dynamics of heavily charged Morse clusters enabled us to vary the range of the pair potential and of the fissibility parameter, which results in distinct fragmentation patterns and in the angular distributions of the fragments. The Rayleigh instability limit separates between nearly binary (or tertiary) spatially unisotropic fission and spatially isotropic Coulomb explosion into a large number of small, ionic fragments. Implications are addressed for a broad spectrum of dynamics in chemical physics, radiation physics of ultracold gases, and biophysics, involving the fission of clusters and droplets, the realization of Coulomb explosion of molecular clusters, the isotropic expansion of optical molasses, and the Coulomb instability of “isolated” proteins. PMID:12093910

Indirect double photoionization of water

NASA Astrophysics Data System (ADS)

Resccigno, T. N.; Sann, H.; Orel, A. E.; Dörner, R.

2011-05-01

The vertical double ionization thresholds of small molecules generally lie above the dissociation limits corresponding to formation of two singly charged fragments. This gives the possibility of populating singly charged molecular ions by photoionization in the Franck-Condon region at energies below the lowest dication state, but above the dissociation limit into two singly charged fragment ions. This process can produce a superexcited neutral fragment that autoionizes at large internuclear separation. We study this process in water, where absorption of a photon produces an inner-shell excited state of H2O+ that fragments to H++OH*. The angular distribution of secondary electrons produced by OH* when it autoionizes produces a characteristic asymmetric pattern that reveals the distance, and therefore the time, at which the decay takes place. LBNL, Berkeley, CA, J. W. Goethe Universität, Frankfurt, Germany. Work performed under auspices of US DOE and supported by OBES, Div. of Chemical Sciences.

Aggregation-fragmentation-diffusion model for trail dynamics

DOE PAGES

Kawagoe, Kyle; Huber, Greg; Pradas, Marc; ...

2017-07-21

We investigate statistical properties of trails formed by a random process incorporating aggregation, fragmentation, and diffusion. In this stochastic process, which takes place in one spatial dimension, two neighboring trails may combine to form a larger one, and also one trail may split into two. In addition, trails move diffusively. The model is defined by two parameters which quantify the fragmentation rate and the fragment size. In the long-time limit, the system reaches a steady state, and our focus is the limiting distribution of trail weights. We find that the density of trail weight has power-law tail P(w)~w –γ formore » small weight w. We obtain the exponent γ analytically and find that it varies continuously with the two model parameters. In conclusion, the exponent γ can be positive or negative, so that in one range of parameters small-weight trails are abundant and in the complementary range they are rare.« less

Long generation time delays the genetic response to habitat fragmentation in the threatened florida sand skink

USGS Publications Warehouse

McCoy, E.D.; Richmond, J.Q.; Mushinsky, H.R.; Britt, E.J.; Godley, J.S.

2010-01-01

A recent study showed that populations of the threatened Florida Sand Skink had limited loss of genetic diversity over the past 60 yr as a consequence of anthropogenic fragmentation. This study assumed that 60 yr represents 3037 generations for the Florida Sand Skink, but a new evaluation of markrecapture data shows that 60 yr represents only about 15 generations. This result suggests that too little time may have passed to observe the full genetic consequences of contemporary anthropogenic fragmentation in the Florida Sand Skink and reinforces similar results from other species. We suggest that snapshots of existing genetic variability in fragmented populations are limited in their ability to predict the evolutionary fate of a species unless life-history attributes of the organism are taken into account. Copyright 2010 Society for the Study of Amphibians and Reptiles.

Measuring Sperm DNA Fragmentation and Clinical Outcomes of Medically Assisted Reproduction: A Systematic Review and Meta-Analysis.

PubMed

Cissen, Maartje; Wely, Madelon van; Scholten, Irma; Mansell, Steven; Bruin, Jan Peter de; Mol, Ben Willem; Braat, Didi; Repping, Sjoerd; Hamer, Geert

2016-01-01

Sperm DNA fragmentation has been associated with reduced fertilization rates, embryo quality, pregnancy rates and increased miscarriage rates. Various methods exist to test sperm DNA fragmentation such as the sperm chromatin structure assay (SCSA), the sperm chromatin dispersion (SCD) test, the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labelling (TUNEL) assay and the single cell gel electrophoresis (Comet) assay. We performed a systematic review and meta-analysis to assess the value of measuring sperm DNA fragmentation in predicting chance of ongoing pregnancy with IVF or ICSI. Out of 658 unique studies, 30 had extractable data and were thus included in the meta-analysis. Overall, the sperm DNA fragmentation tests had a reasonable to good sensitivity. A wide variety of other factors may also affect the IVF/ICSI outcome, reflected by limited to very low specificity. The constructed hierarchical summary receiver operating characteristic (HSROC) curve indicated a fair discriminatory capacity of the TUNEL assay (area under the curve (AUC) of 0.71; 95% CI 0.66 to 0.74) and Comet assay (AUC of 0.73; 95% CI 0.19 to 0.97). The SCSA and the SCD test had poor predictive capacity. Importantly, for the TUNEL assay, SCD test and Comet assay, meta-regression showed no differences in predictive value between IVF and ICSI. For the SCSA meta-regression indicated the predictive values for IVF and ICSI were different. The present review suggests that current sperm DNA fragmentation tests have limited capacity to predict the chance of pregnancy in the context of MAR. Furthermore, sperm DNA fragmentation tests have little or no difference in predictive value between IVF and ICSI. At this moment, there is insufficient evidence to recommend the routine use of sperm DNA fragmentation tests in couples undergoing MAR both for the prediction of pregnancy and for the choice of treatment. Given the significant limitations of the evidence and the methodological weakness and design of the included studies, we do urge for further research on the predictive value of sperm DNA fragmentation for the chance of pregnancy after MAR, also in comparison with other predictors of pregnancy after MAR.

Fragmentation of interstellar clouds and star formation

NASA Technical Reports Server (NTRS)

Silk, J.

1982-01-01

The principal issues are addressed: the fragmentation of molecular clouds into units of stellar mass and the impact of star formation on molecular clouds. The observational evidence for fragmentation is summarized, and the gravitational instability described of a uniform spherical cloud collapsing from rest. The implications are considered of a finite pressure for the minimum fragment mass that is attainable in opacity-limited fragmentation. The role of magnetic fields is discussed in resolving the angular momentum problem and in making the collapse anisotropic, with notable consequences for fragmentation theory. Interactions between fragments are described, with emphasis on the effect of protostellar winds on the ambient cloud matter and on inhibiting further star formation. Such interactions are likely to have profound consequences for regulating the rate of star formation and on the energetics and dynamics of molecular clouds.

Achieving climate connectivity in a fragmented landscape

PubMed Central

Lawler, Joshua J.; McRae, Brad H.; Nuñez, Tristan A.; Theobald, David M.

2016-01-01

The contiguous United States contains a disconnected patchwork of natural lands. This fragmentation by human activities limits species’ ability to track suitable climates as they rapidly shift. However, most models that project species movement needs have not examined where fragmentation will limit those movements. Here, we quantify climate connectivity, the capacity of landscape configuration to allow species movement in the face of dynamically shifting climate. Using this metric, we assess to what extent habitat fragmentation will limit species movements in response to climate change. We then evaluate how creating corridors to promote climate connectivity could potentially mitigate these restrictions, and we assess where strategies to increase connectivity will be most beneficial. By analyzing fragmentation patterns across the contiguous United States, we demonstrate that only 41% of natural land area retains enough connectivity to allow plants and animals to maintain climatic parity as the climate warms. In the eastern United States, less than 2% of natural area is sufficiently connected. Introducing corridors to facilitate movement through human-dominated regions increases the percentage of climatically connected natural area to 65%, with the most impactful gains in low-elevation regions, particularly in the southeastern United States. These climate connectivity analyses allow ecologists and conservation practitioners to determine the most effective regions for increasing connectivity. More importantly, our findings demonstrate that increasing climate connectivity is critical for allowing species to track rapidly changing climates, reconfiguring habitats to promote access to suitable climates. PMID:27298349

Achieving climate connectivity in a fragmented landscape.

PubMed

McGuire, Jenny L; Lawler, Joshua J; McRae, Brad H; Nuñez, Tristan A; Theobald, David M

2016-06-28

The contiguous United States contains a disconnected patchwork of natural lands. This fragmentation by human activities limits species' ability to track suitable climates as they rapidly shift. However, most models that project species movement needs have not examined where fragmentation will limit those movements. Here, we quantify climate connectivity, the capacity of landscape configuration to allow species movement in the face of dynamically shifting climate. Using this metric, we assess to what extent habitat fragmentation will limit species movements in response to climate change. We then evaluate how creating corridors to promote climate connectivity could potentially mitigate these restrictions, and we assess where strategies to increase connectivity will be most beneficial. By analyzing fragmentation patterns across the contiguous United States, we demonstrate that only 41% of natural land area retains enough connectivity to allow plants and animals to maintain climatic parity as the climate warms. In the eastern United States, less than 2% of natural area is sufficiently connected. Introducing corridors to facilitate movement through human-dominated regions increases the percentage of climatically connected natural area to 65%, with the most impactful gains in low-elevation regions, particularly in the southeastern United States. These climate connectivity analyses allow ecologists and conservation practitioners to determine the most effective regions for increasing connectivity. More importantly, our findings demonstrate that increasing climate connectivity is critical for allowing species to track rapidly changing climates, reconfiguring habitats to promote access to suitable climates.

Limiting fragmentation from scale-invariant merging of fast partons

NASA Astrophysics Data System (ADS)

Bialas, A.; Bzdak, A.; Peschanski, R.

2008-07-01

Exploiting the idea that the fast partons of an energetic projectile can be treated as sources of colour radiation interpreted as wee partons, it is shown that the recently observed property of extended limiting fragmentation implies a scaling law for the rapidity distribution of fast partons. This leads to a picture of a self-similar process where, for fixed total rapidity Y, the sources merge with probability varying as 1 / y.

Pollen limitation and reduced reproductive success are associated with local genetic effects in Prunus virginiana, a widely distributed self-incompatible shrub.

PubMed

Suarez-Gonzalez, Adriana; Good, Sara V

2014-03-01

A vast quantity of empirical evidence suggests that insufficient quantity or quality of pollen may lead to a reduction in fruit set, in particular for self-incompatible species. This study uses an integrative approach that combines field research with marker gene analysis to understand the factors affecting reproductive success in a widely distributed self-incompatible species, Prunus virginiana (Rosaceae). Twelve patches of P. virginiana distributed within three populations that differed in degree of disturbance were examined. Two of the sites were small (7-35 km(2)) remnants of forest in an intensively used agricultural landscape, while the third was continuous (350 km(2)) and less disturbed. Field studies (natural and hand cross-pollinations) were combined with marker gene analyses (microsatellites and S-locus) in order to explore potential factors affecting pollen delivery and consequently reproductive success at landscape (between populations) and fine scales (within populations). Reductions in reproductive output were found in the two fragments compared with the continuous population, and suggest that pollen is an important factor limiting fruit production. Genetic analyses carried out in one of the fragments and in the continuous site suggest that even though S-allele diversity is high in both populations, the fragment exhibits an increase in biparental inbreeding and correlated paternity. The increase in biparental inbreeding in the fragment is potentially attributable to variation in the density of individuals and/or the spatial distribution of genotypes among populations, both of which could alter mating dynamics. By using a novel integrative approach, this study shows that even though P. virginiana is a widespread species, fragmented populations can experience significant reductions in fruit set and pollen limitation in the field. Deatiled examination of one fragmented population suggests that these linitations may be explained by an increase in biparental inbreeding, correlated paternity and fine-scale genetic structure. The consistency of the field and fine-scale genetic analyses, and the consistency of the results within patches and across years, suggest that these are important processes driving pollen limitation in the fragment.

Pollen limitation and reduced reproductive success are associated with local genetic effects in Prunus virginiana, a widely distributed self-incompatible shrub

PubMed Central

Suarez-Gonzalez, Adriana; Good, Sara V.

2014-01-01

Background and Aims A vast quantity of empirical evidence suggests that insufficient quantity or quality of pollen may lead to a reduction in fruit set, in particular for self-incompatible species. This study uses an integrative approach that combines field research with marker gene analysis to understand the factors affecting reproductive success in a widely distributed self-incompatible species, Prunus virginiana (Rosaceae). Methods Twelve patches of P. virginiana distributed within three populations that differed in degree of disturbance were examined. Two of the sites were small (7–35 km2) remnants of forest in an intensively used agricultural landscape, while the third was continuous (350 km2) and less disturbed. Field studies (natural and hand cross-pollinations) were combined with marker gene analyses (microsatellites and S-locus) in order to explore potential factors affecting pollen delivery and consequently reproductive success at landscape (between populations) and fine scales (within populations). Key Results Reductions in reproductive output were found in the two fragments compared with the continuous population, and suggest that pollen is an important factor limiting fruit production. Genetic analyses carried out in one of the fragments and in the continuous site suggest that even though S-allele diversity is high in both populations, the fragment exhibits an increase in biparental inbreeding and correlated paternity. The increase in biparental inbreeding in the fragment is potentially attributable to variation in the density of individuals and/or the spatial distribution of genotypes among populations, both of which could alter mating dynamics. Conclusions By using a novel integrative approach, this study shows that even though P. virginiana is a widespread species, fragmented populations can experience significant reductions in fruit set and pollen limitation in the field. Deatiled examination of one fragmented population suggests that these linitations may be explained by an increase in biparental inbreeding, correlated paternity and fine-scale genetic structure. The consistency of the field and fine-scale genetic analyses, and the consistency of the results within patches and across years, suggest that these are important processes driving pollen limitation in the fragment. PMID:24327534

Meta-analysis of susceptibility of woody plants to loss of genetic diversity through habitat fragmentation.

PubMed

Vranckx, Guy; Jacquemyn, Hans; Muys, Bart; Honnay, Olivier

2012-04-01

Shrubs and trees are assumed less likely to lose genetic variation in response to habitat fragmentation because they have certain life-history characteristics such as long lifespans and extensive pollen flow. To test this assumption, we conducted a meta-analysis with data on 97 woody plant species derived from 98 studies of habitat fragmentation. We measured the weighted response of four different measures of population-level genetic diversity to habitat fragmentation with Hedge's d and Spearman rank correlation. We tested whether the genetic response to habitat fragmentation was mediated by life-history traits (longevity, pollination mode, and seed dispersal vector) and study characteristics (genetic marker and plant material used). For both tests of effect size habitat fragmentation was associated with a substantial decrease in expected heterozygosity, number of alleles, and percentage of polymorphic loci, whereas the population inbreeding coefficient was not associated with these measures. The largest proportion of variation among effect sizes was explained by pollination mechanism and by the age of the tissue (progeny or adult) that was genotyped. Our primary finding was that wind-pollinated trees and shrubs appeared to be as likely to lose genetic variation as insect-pollinated species, indicating that severe habitat fragmentation may lead to pollen limitation and limited gene flow. In comparison with results of previous meta-analyses on mainly herbaceous species, we found trees and shrubs were as likely to have negative genetic responses to habitat fragmentation as herbaceous species. We also found that the genetic variation in offspring was generally less than that of adult trees, which is evidence of a genetic extinction debt and probably reflects the genetic diversity of the historical, less-fragmented landscape. ©2011 Society for Conservation Biology.

Mass spectrometry for fragment screening.

PubMed

Chan, Daniel Shiu-Hin; Whitehouse, Andrew J; Coyne, Anthony G; Abell, Chris

2017-11-08

Fragment-based approaches in chemical biology and drug discovery have been widely adopted worldwide in both academia and industry. Fragment hits tend to interact weakly with their targets, necessitating the use of sensitive biophysical techniques to detect their binding. Common fragment screening techniques include differential scanning fluorimetry (DSF) and ligand-observed NMR. Validation and characterization of hits is usually performed using a combination of protein-observed NMR, isothermal titration calorimetry (ITC) and X-ray crystallography. In this context, MS is a relatively underutilized technique in fragment screening for drug discovery. MS-based techniques have the advantage of high sensitivity, low sample consumption and being label-free. This review highlights recent examples of the emerging use of MS-based techniques in fragment screening. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Repositioning the substrate activity screening (SAS) approach as a fragment-based method for identification of weak binders.

PubMed

Gladysz, Rafaela; Cleenewerck, Matthias; Joossens, Jurgen; Lambeir, Anne-Marie; Augustyns, Koen; Van der Veken, Pieter

2014-10-13

Fragment-based drug discovery (FBDD) has evolved into an established approach for "hit" identification. Typically, most applications of FBDD depend on specialised cost- and time-intensive biophysical techniques. The substrate activity screening (SAS) approach has been proposed as a relatively cheap and straightforward alternative for identification of fragments for enzyme inhibitors. We have investigated SAS for the discovery of inhibitors of oncology target urokinase (uPA). Although our results support the key hypotheses of SAS, we also encountered a number of unreported limitations. In response, we propose an efficient modified methodology: "MSAS" (modified substrate activity screening). MSAS circumvents the limitations of SAS and broadens its scope by providing additional fragments and more coherent SAR data. As well as presenting and validating MSAS, this study expands existing SAR knowledge for the S1 pocket of uPA and reports new reversible and irreversible uPA inhibitor scaffolds. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Bremsstrahlung from colour charges as a source of soft particle production in hadronic collisions

NASA Astrophysics Data System (ADS)

Bialas, A.; Jezabek, M.

2004-06-01

It is proposed that soft particle production in hadronic collisions is dominated by multiple gluon exchanges between partons from the colliding hadrons, followed by radiation of hadronic clusters from the coloured partons distributed uniformly in rapidity. This explains naturally two dominant features of the data: (a) the linear increase of rapidity spectra in the regions of limiting fragmentation and, (b) the proportionality between the increasing width of the limiting fragmentation region and the height of the central plateau.

Characteristics of Whipple Shield Performance in the Shatter Regime

NASA Technical Reports Server (NTRS)

Ryan, S.; Bjorkman, M.; Christiansen, E. L.

2010-01-01

Ballistic limit equations define the failure of metallic Whipple shields in three parts: low velocity, shatter, and hypervelocity. Failure limits in the shatter regime are based on a linear interpolation between the onset of projectile fragmentation, and impulsive rupture of the shield rear wall. A series of hypervelocity impact tests have been performed on aluminum alloy Whipple shields to investigate failure mechanisms and performance limits in the shatter regime. Test results demonstrated a more rapid increase in performance than predicted by the latest iteration of the JSC Whipple shield BLE following the onset of projectile fragmentation. This increase in performance was found to level out between 4.0-5.0 km/s, with a subsequent decrease in performance for velocities up to 6.0 km/s. For a detached spall failure criterion, the failure limit was found to continually decrease up to a velocity of 7.0 km/s, substantially varying from the BLE, while for perforation-based failure an increase in performance was observed. An existing phenomenological ballistic limit curve was found to provide a more accurate reproduction of shield behavior that the BLE, however a number of underlying assumptions such as the occurrence of complete projectile fragmentation and the effect on performance of incipient projectile melt were found to be inaccurate. A cratering relationship based on the largest residual fragment size has been derived for application at velocities between 3.0-4.0 km/s, and was shown to accurately reproduce the trends of the experimental data. Further investigation is required to allow a full analytical description of shatter regime performance for metallic Whipple shields.

Shifted termination assay (STA) fragment analysis to detect BRAF V600 mutations in papillary thyroid carcinomas

PubMed Central

2013-01-01

Background BRAF mutation is an important diagnostic and prognostic marker in patients with papillary thyroid carcinoma (PTC). To be applicable in clinical laboratories with limited equipment, diverse testing methods are required to detect BRAF mutation. Methods A shifted termination assay (STA) fragment analysis was used to detect common V600 BRAF mutations in 159 PTCs with DNAs extracted from formalin-fixed paraffin-embedded tumor tissue. The results of STA fragment analysis were compared to those of direct sequencing. Serial dilutions of BRAF mutant cell line (SNU-790) were used to calculate limit of detection (LOD). Results BRAF mutations were detected in 119 (74.8%) PTCs by STA fragment analysis. In direct sequencing, BRAF mutations were observed in 118 (74.2%) cases. The results of STA fragment analysis had high correlation with those of direct sequencing (p < 0.00001, κ = 0.98). The LOD of STA fragment analysis and direct sequencing was 6% and 12.5%, respectively. In PTCs with pT3/T4 stages, BRAF mutation was observed in 83.8% of cases. In pT1/T2 carcinomas, BRAF mutation was detected in 65.9% and this difference was statistically significant (p = 0.007). Moreover, BRAF mutation was more frequent in PTCs with extrathyroidal invasion than tumors without extrathyroidal invasion (84.7% versus 62.2%, p = 0.001). To prepare and run the reactions, direct sequencing required 450 minutes while STA fragment analysis needed 290 minutes. Conclusions STA fragment analysis is a simple and sensitive method to detect BRAF V600 mutations in formalin-fixed paraffin-embedded clinical samples. Virtual Slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/5684057089135749 PMID:23883275

Relativistic corrections to heavy quark fragmentation to S-wave heavy mesons

NASA Astrophysics Data System (ADS)

Sang, Wen-Long; Yang, Lan-Fei; Chen, Yu-Qi

2009-07-01

The relativistic corrections of order v2 to the fragmentation functions for the heavy quark to S-wave heavy quarkonia are calculated in the framework of the nonrelativistic quantum chromodynamics factorization formula. We derive the fragmentation functions by using the Collins-Soper definition in both the Feynman gauge and the axial gauge. We also extract them through the process Z0→Hq qmacr in the limit MZ/m→∞. We find that all results obtained by these two different methods and in different gauges are the same. We estimate the relative size of the relativistic corrections to the fragmentation functions.

Relativistic corrections to heavy quark fragmentation to S-wave heavy mesons

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sang Wenlong; Yang Lanfei; Chen Yuqi

The relativistic corrections of order v{sup 2} to the fragmentation functions for the heavy quark to S-wave heavy quarkonia are calculated in the framework of the nonrelativistic quantum chromodynamics factorization formula. We derive the fragmentation functions by using the Collins-Soper definition in both the Feynman gauge and the axial gauge. We also extract them through the process Z{sup 0}{yields}Hqq in the limit M{sub Z}/m{yields}{infinity}. We find that all results obtained by these two different methods and in different gauges are the same. We estimate the relative size of the relativistic corrections to the fragmentation functions.

Computational Fragment-Based Drug Design: Current Trends, Strategies, and Applications.

PubMed

Bian, Yuemin; Xie, Xiang-Qun Sean

2018-04-09

Fragment-based drug design (FBDD) has become an effective methodology for drug development for decades. Successful applications of this strategy brought both opportunities and challenges to the field of Pharmaceutical Science. Recent progress in the computational fragment-based drug design provide an additional approach for future research in a time- and labor-efficient manner. Combining multiple in silico methodologies, computational FBDD possesses flexibilities on fragment library selection, protein model generation, and fragments/compounds docking mode prediction. These characteristics provide computational FBDD superiority in designing novel and potential compounds for a certain target. The purpose of this review is to discuss the latest advances, ranging from commonly used strategies to novel concepts and technologies in computational fragment-based drug design. Particularly, in this review, specifications and advantages are compared between experimental and computational FBDD, and additionally, limitations and future prospective are discussed and emphasized.

Preliminary insights into a model for mafic magma fragmentation

NASA Astrophysics Data System (ADS)

Edwards, Matt; Pioli, Laura; Andronico, Daniele; Cristaldi, Antonio; Scollo, Simona

2017-04-01

Fragmentation of mafic magmas remains a poorly understood process despite the common occurrence of low viscosity explosive eruptions. In fact, it has been commonly overlooked based on the assumption that low viscosity magmas have very limited explosivity and low potential to undergo brittle fragmentation. However, it is now known that highly explosive, ash forming eruptions can be relatively frequent at several mafic volcanoes. Three questions arise due to this - What is the specific fragmentation mechanism occuring in these eruptions? What are the primary factors controlling fragmentation efficiency? Can a link between eruption style and fragmentation efficiency be quantified? We addressed these questions by coupling theoretical observations and field analysis of the recent May 2016 eruption at Mount Etna volcano. Within this complex 10-day event three paroxysmal episodes of pulsating basaltic lava jets alternating with small lava flows were recorded from a vent within the Voragine crater. The associated plumes which were produced deposited tephra along narrow axes to the east and south east. Sampling was done on the deposits associated with the first two plumes and the third one. We briefly characterise the May 2016 eruption by assessing plume height, eruption phases, total erupted masses and fallout boundaries and comparing them to previous eruptions. We also analyse the total grainsize distribution (TGSD) of the scoria particles formed in the jets. Conventional methods for obtaining grainsize and total distributions of an eruption are based on mass and provide limited information on fragmentation though. For this reason, the TGSD was assessed by coupling particle analyser data and conventional sieving data to assess both particle size and number of particle distributions with better precision. This allowed for more accurate testing of several existing models describing the shape of the TGSD. Coupled further with observations on eruption dynamics and eruption phase durations obtained from the network of fixed INGV cameras, early insight into possible links between fragmentation and eruption conditions are identified. A link between fragmentation and magma properties is also examined. We discuss the relationship between the conventional and new analytical methods and their potential in unraveling key information on the fragmentation process and analyse how the dataset on the May eruption can be modelled with the current fragmentation theories. Finally, we suggest the systematic use of a comprehensive TGSD dataset to develop a fragmentation model for mafic eruptions.

The properties of retrieval cues constrain the picture superiority effect.

PubMed

Weldon, M S; Roediger, H L; Challis, B H

1989-01-01

In three experiments, we examined why pictures are remembered better than words on explicit memory tests like recall and recognition, whereas words produce more priming than pictures on some implicit tests, such as word-fragment and word-stem completion (e.g., completing -l-ph-nt or ele----- as elephant). One possibility is that pictures are always more accessible than words if subjects are given explicit retrieval instructions. An alternative possibility is that the properties of the retrieval cues themselves constrain the retrieval processes engaged; word fragments might induce data-driven (perceptually based) retrieval, which favors words regardless of the retrieval instructions. Experiment 1 demonstrated that words were remembered better than pictures on both the word-fragment and word-stem completion tasks under both implicit and explicit retrieval conditions. In Experiment 2, pictures were recalled better than words with semantically related extralist cues. In Experiment 3, when semantic cues were combined with word fragments, pictures and words were recalled equally well under explicit retrieval conditions, but words were superior to pictures under implicit instructions. Thus, the inherently data-limited properties of fragmented words limit their use in accessing conceptual codes. Overall, the results indicate that retrieval operations are largely determined by properties of the retrieval cues under both implicit and explicit retrieval conditions.

Integrating the Engineering Curriculum through the Synthesis and Design Studio

ERIC Educational Resources Information Center

Kellam, Nadia; Walther, Joachim; Costantino, Tracie; Cramond, Bonnie

2013-01-01

Traditional curricular approaches within engineering education tend to be fragmented, with opportunities for content- and meta-level synthesis being mostly limited to freshman and senior year design courses. In this paper, we are proposing a curricular model, the Synthesis and Design Studio, to combat the tendency towards fragmented curricula. The…

Kohonen and counterpropagation neural networks applied for mapping and interpretation of IR spectra.

PubMed

Novic, Marjana

2008-01-01

The principles of learning strategy of Kohonen and counterpropagation neural networks are introduced. The advantages of unsupervised learning are discussed. The self-organizing maps produced in both methods are suitable for a wide range of applications. Here, we present an example of Kohonen and counterpropagation neural networks used for mapping, interpretation, and simulation of infrared (IR) spectra. The artificial neural network models were trained for prediction of structural fragments of an unknown compound from its infrared spectrum. The training set contained over 3,200 IR spectra of diverse compounds of known chemical structure. The structure-spectra relationship was encompassed by the counterpropagation neural network, which assigned structural fragments to individual compounds within certain probability limits, assessed from the predictions of test compounds. The counterpropagation neural network model for prediction of fragments of chemical structure is reversible, which means that, for a given structural domain, limited to the training data set in the study, it can be used to simulate the IR spectrum of a chemical defined with a set of structural fragments.

A Case of Nonunion Avulsion Fracture of the Anterior Tibial Eminence

PubMed Central

Atsumi, Satoru; Arai, Yuji; Nakagawa, Shuji; Inoue, Hiroaki; Ikoma, Kazuya; Fujiwara, Hiroyoshi; Kubo, Toshikazu

2016-01-01

Avulsion fracture of the anterior tibial eminence is an uncommon injury. If bone union does not occur, knee extension will be limited by impingement of the avulsed fragment and knee instability will be induced by dysfunction of the anterior cruciate ligament (ACL). This report describes a 55-year-old woman who experienced an avulsion fracture of the right anterior tibial eminence during recreational skiing. Sixteen months later, she presented at our hospital with limitation of right knee extension. Plain radiography showed nonunion of the avulsion fracture region, and arthroscopy showed that the avulsed fragment impinged the femoral intercondylar notch during knee extension. The anterior region of the bony fragment was debrided arthroscopically until the knee could be extended completely. There was no subsequent instability, and the patient was able to climb a mountain 6 months after surgery. These findings indicate that arthroscopic debridement of an avulsed fragment for nonunion of an avulsion fracture of the anterior tibial eminence is a minimally invasive and effective treatment for middle-aged and elderly patients with a low level of sports activity. PMID:27119035

An innovative platform for quick and flexible joining of assorted DNA fragments

DOE PAGES

De Paoli, Henrique Cestari; Tuskan, Gerald A.; Yang, Xiaohan

2016-01-13

Successful synthetic biology efforts rely on conceptual and experimental designs in combination with testing of multi-gene constructs. Despite recent progresses, several limitations still hinder the ability to flexibly assemble and collectively share different types of DNA segments. We describe an advanced system for joining DNA fragments from a universal library that automatically maintains open reading frames (ORFs) and does not require linkers, adaptors, sequence homology, amplification or mutation (domestication) of fragments in order to work properly. Moreover, we find that this system, which is enhanced by a unique buffer formulation, provides unforeseen capabilities for testing, and sharing, complex multi-gene circuitrymore » assembled from different DNA fragments.« less

Anthropogenic fragmentation may not alter pre-existing patterns of genetic diversity and differentiation in perennial shrubs.

PubMed

Llorens, Tanya M; Ayre, David J; Whelan, Robert J

2018-04-01

Many plant species have pollination and seed dispersal systems and evolutionary histories that have produced strong genetic structuring. These genetic patterns may be consistent with expectations following recent anthropogenic fragmentation, making it difficult to detect fragmentation effects if no prefragmentation genetic data are available. We used microsatellite markers to investigate whether severe habitat fragmentation may have affected the structure and diversity of populations of the endangered Australian bird-pollinated shrub Grevillea caleyi R.Br., by comparing current patterns of genetic structure and diversity with those of the closely related G. longifolia R.Br. that has a similar life history but has not experienced anthropogenic fragmentation. Grevillea caleyi and G. longifolia showed similar and substantial population subdivision at all spatial levels (global F' ST  = 0.615 and 0.454; S p  = 0.039 and 0.066), marked isolation by distance and large heterozygous deficiencies. These characteristics suggest long-term effects of inbreeding in self-compatible species that have poor seed dispersal, limited connectivity via pollen flow and undergo population bottlenecks because of periodic fires. Highly structured allele size distributions, most notably in G. caleyi, imply historical processes of drift and mutation were important in isolated subpopulations. Genetic diversity did not vary with population size but was lower in more isolated populations for both species. Through this comparison, we reject the hypothesis that anthropogenic fragmentation has impacted substantially on the genetic composition or structure of G. caleyi populations. Our results suggest that highly self-compatible species with limited dispersal may be relatively resilient to the genetic changes predicted to follow habitat fragmentation. © 2018 John Wiley & Sons Ltd.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Jiang; Malmirchegini, G. Reza; Clubb, Robert T.

Native mass spectrometry (MS) has become an invaluable tool for the characterization of proteins and non-covalent protein complexes under near physiological solution conditions. Here we report the structural characterization of human hemoglobin (Hb), a 64 kDa oxygen-transporting protein complex, by high resolution native top-down mass spectrometry using electrospray ionization (ESI) and a 15-Tesla Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. Native MS preserves the non-covalent interactions between the globin subunits, and electron capture dissociation (ECD) produces fragments directly from the intact Hb complex without dissociating the subunits. Using activated ion ECD, we observe the gradual unfolding process of themore » Hb complex in the gas phase. Without protein ion activation, the native Hb shows very limited ECD fragmentation from the N-termini, suggesting a tightly packed structure of the native complex and therefore low fragmentation efficiency. Precursor ion activation allows steady increase of N-terminal fragment ions, while the C-terminal fragments remain limited (38 c ions and 4 z ions on the α chain; 36 c ions and 2 z ions on the β chain). This ECD fragmentation pattern suggests that upon activation, the Hb complex starts to unfold from the N-termini of both subunits, whereas the C-terminal regions and therefore the potential regions involved in the subunit binding interactions remain intact. ECD-MS of the Hb dimer show similar fragmentation patterns as the Hb tetramer, providing further evidence for the hypothesized unfolding process of the Hb complex in the gas phase. Native top-down ECD-MS allows efficient probing of the Hb complex structure and the subunit binding interactions in the gas phase. Finally, it may provide a fast and effective means to probe the structure of novel protein complexes that are intractable to traditional structural characterization tools.« less

Cracks dynamics under tensional stress - a DEM approach

NASA Astrophysics Data System (ADS)

Debski, Wojciech; Klejment, Piotr; Kosmala, Alicja; Foltyn, Natalia; Szpindler, Maciej

2017-04-01

Breaking and fragmentation of solid materials is an extremely complex process involving scales ranging from an atomic scale (breaking inter-atomic bounds) up to thousands of kilometers in case of catastrophic earthquakes (in energy scale it ranges from single eV up to 1024 J). Such a large scale span of breaking processes opens lot of questions like, for example, scaling of breaking processes, existence of factors controlling final size of broken area, existence of precursors, dynamics of fragmentation, to name a few. The classical approach to study breaking process at seismological scales, i.e., physical processes in earthquake foci, is essentially based on two factors: seismic data (mostly) and the continuum mechanics (including the linear fracture mechanics). Such approach has been gratefully successful in developing kinematic (first) and dynamic (recently) models of seismic rupture and explaining many of earthquake features observed all around the globe. However, such approach will sooner or latter face a limitation due to a limited information content of seismic data and inherit limitations of the fracture mechanics principles. A way of avoiding this expected limitation is turning an attention towards a well established in physics method of computational simulations - a powerful branch of contemporary physics. In this presentation we discuss preliminary results of analysis of fracturing dynamics under external tensional forces using the Discrete Element Method approach. We demonstrate that even under a very simplified tensional conditions, the fragmentation dynamics is a very complex process, including multi-fracturing, spontaneous fracture generation and healing, etc. We also emphasis a role of material heterogeneity on the fragmentation process.

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF.

PubMed

Ellis, W C; Mahlooji, M; Lascano, C E; Matis, J H

2005-07-01

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.

An accurate algorithm for the detection of DNA fragments from dilution pool sequencing experiments.

PubMed

Bansal, Vikas

2018-01-01

The short read lengths of current high-throughput sequencing technologies limit the ability to recover long-range haplotype information. Dilution pool methods for preparing DNA sequencing libraries from high molecular weight DNA fragments enable the recovery of long DNA fragments from short sequence reads. These approaches require computational methods for identifying the DNA fragments using aligned sequence reads and assembling the fragments into long haplotypes. Although a number of computational methods have been developed for haplotype assembly, the problem of identifying DNA fragments from dilution pool sequence data has not received much attention. We formulate the problem of detecting DNA fragments from dilution pool sequencing experiments as a genome segmentation problem and develop an algorithm that uses dynamic programming to optimize a likelihood function derived from a generative model for the sequence reads. This algorithm uses an iterative approach to automatically infer the mean background read depth and the number of fragments in each pool. Using simulated data, we demonstrate that our method, FragmentCut, has 25-30% greater sensitivity compared with an HMM based method for fragment detection and can also detect overlapping fragments. On a whole-genome human fosmid pool dataset, the haplotypes assembled using the fragments identified by FragmentCut had greater N50 length, 16.2% lower switch error rate and 35.8% lower mismatch error rate compared with two existing methods. We further demonstrate the greater accuracy of our method using two additional dilution pool datasets. FragmentCut is available from https://bansal-lab.github.io/software/FragmentCut. vibansal@ucsd.edu. Supplementary data are available at Bioinformatics online. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Research on the fragment impact resistance of a composite mast

NASA Astrophysics Data System (ADS)

Zhang, Wei; Wu, Chao; Yang, Wenshan; Xu, Shanshan; Ren, Shaofei

2011-09-01

Due to the unique structural mode and material property of a composite sandwich plate, related research such as fragment impact resistance of a composite mast is short of publication and urgent in this field. In this paper, the commonly accepted sandwich core board theory was modified. Damage caused by a fragment attack was simulated onto a sandwich plate model built with solid and shell elements. It was shown that shear failure and vast matrix cracking are the main reasons for outer coat damage, and tension failure and partial matrix cracking are the cause for inner coat damage. Additionally, according to complexities in actual sea battles, different work conditions of missile attacks were set. Ballistic limit values of different fragment sizes were also obtained, which provides references for enhancing the fragment impact resistance of a composite mast.

Characterization of protein N-glycosylation by tandem mass spectrometry using complementary fragmentation techniques

DOE PAGES

Ford, Kristina L.; Zeng, Wei; Heazlewood, Joshua L.; ...

2015-08-28

The analysis of post-translational modifications (PTMs) by proteomics is regarded as a technically challenging undertaking. While in recent years approaches to examine and quantify protein phosphorylation have greatly improved, the analysis of many protein modifications, such as glycosylation, are still regarded as problematic. Limitations in the standard proteomics workflow, such as use of suboptimal peptide fragmentation methods, can significantly prevent the identification of glycopeptides. The current generation of tandem mass spectrometers has made available a variety of fragmentation options, many of which are becoming standard features on these instruments. Lastly, we have used three common fragmentation techniques, namely CID, HCD,more » and ETD, to analyze a glycopeptide and highlight how an integrated fragmentation approach can be used to identify the modified residue and characterize the N-glycan on a peptide.« less

Critical Features of Fragment Libraries for Protein Structure Prediction

PubMed Central

dos Santos, Karina Baptista

2017-01-01

The use of fragment libraries is a popular approach among protein structure prediction methods and has proven to substantially improve the quality of predicted structures. However, some vital aspects of a fragment library that influence the accuracy of modeling a native structure remain to be determined. This study investigates some of these features. Particularly, we analyze the effect of using secondary structure prediction guiding fragments selection, different fragments sizes and the effect of structural clustering of fragments within libraries. To have a clearer view of how these factors affect protein structure prediction, we isolated the process of model building by fragment assembly from some common limitations associated with prediction methods, e.g., imprecise energy functions and optimization algorithms, by employing an exact structure-based objective function under a greedy algorithm. Our results indicate that shorter fragments reproduce the native structure more accurately than the longer. Libraries composed of multiple fragment lengths generate even better structures, where longer fragments show to be more useful at the beginning of the simulations. The use of many different fragment sizes shows little improvement when compared to predictions carried out with libraries that comprise only three different fragment sizes. Models obtained from libraries built using only sequence similarity are, on average, better than those built with a secondary structure prediction bias. However, we found that the use of secondary structure prediction allows greater reduction of the search space, which is invaluable for prediction methods. The results of this study can be critical guidelines for the use of fragment libraries in protein structure prediction. PMID:28085928

Critical Features of Fragment Libraries for Protein Structure Prediction.

PubMed

Trevizani, Raphael; Custódio, Fábio Lima; Dos Santos, Karina Baptista; Dardenne, Laurent Emmanuel

2017-01-01

The use of fragment libraries is a popular approach among protein structure prediction methods and has proven to substantially improve the quality of predicted structures. However, some vital aspects of a fragment library that influence the accuracy of modeling a native structure remain to be determined. This study investigates some of these features. Particularly, we analyze the effect of using secondary structure prediction guiding fragments selection, different fragments sizes and the effect of structural clustering of fragments within libraries. To have a clearer view of how these factors affect protein structure prediction, we isolated the process of model building by fragment assembly from some common limitations associated with prediction methods, e.g., imprecise energy functions and optimization algorithms, by employing an exact structure-based objective function under a greedy algorithm. Our results indicate that shorter fragments reproduce the native structure more accurately than the longer. Libraries composed of multiple fragment lengths generate even better structures, where longer fragments show to be more useful at the beginning of the simulations. The use of many different fragment sizes shows little improvement when compared to predictions carried out with libraries that comprise only three different fragment sizes. Models obtained from libraries built using only sequence similarity are, on average, better than those built with a secondary structure prediction bias. However, we found that the use of secondary structure prediction allows greater reduction of the search space, which is invaluable for prediction methods. The results of this study can be critical guidelines for the use of fragment libraries in protein structure prediction.

Heterogeneous information sharing of sensor information in contested environments

NASA Astrophysics Data System (ADS)

Wampler, Jason A.; Hsieh, Chien; Toth, Andrew; Sheatsley, Ryan

2017-05-01

The inherent nature of unattended sensors makes these devices most vulnerable to detection, exploitation, and denial in contested environments. Physical access is often cited as the easiest way to compromise any device or network. A new mechanism for mitigating these types of attacks developed under the Assistant Secretary of Defense for Research and Engineering, ASD(R and E) project, "Smoke Screen in Cyberspace", was demonstrated in a live, over-the-air experiment. Smoke Screen encrypts, slices up, and disburses redundant fragments of files throughout the network. Recovery is only possible after recovering all fragments and attacking/denying one or more nodes does not limit the availability of other fragment copies in the network. This experiment proved the feasibility of redundant file fragmentation, and is the foundation for developing sophisticated methods to blacklist compromised nodes, move data fragments from risks of compromise, and forward stored data fragments closer to the anticipated retrieval point. This paper outlines initial results in scalability of node members, fragment size, file size, and performance in a heterogeneous network consisting of the Wireless Network after Next (WNaN) radio and Common Sensor Radio (CSR).

Immobilization of Fab' fragments onto substrate surfaces: A survey of methods and applications.

PubMed

Crivianu-Gaita, Victor; Thompson, Michael

2015-08-15

Antibody immobilization onto surfaces has widespread applications in many different fields. It is desirable to bind antibodies such that their fragment-antigen-binding (Fab) units are oriented away from the surface in order to maximize analyte binding. The immobilization of only Fab' fragments yields benefits over the more traditional whole antibody immobilization technique. Bound Fab' fragments display higher surface densities, yielding a higher binding capacity for the analyte. The nucleophilic sulfide of the Fab' fragments allows for specific orientations to be achieved. For biosensors, this indicates a higher sensitivity and lower detection limit for a target analyte. The last thirty years have shown tremendous progress in the immobilization of Fab' fragments onto gold, Si-based, polysaccharide-based, plastic-based, magnetic, and inorganic surfaces. This review will show the current scope of Fab' immobilization techniques available and illustrate methods employed to minimize non-specific adsorption of undesirables. Furthermore, a variety of examples will be given to show the versatility of immobilized Fab' fragments in different applications and future directions of the field will be addressed, especially regarding biosensors. Copyright © 2015 Elsevier B.V. All rights reserved.

Recombinant PrPSc shares structural features with brain-derived PrPSc: Insights from limited proteolysis.

PubMed

Sevillano, Alejandro M; Fernández-Borges, Natalia; Younas, Neelam; Wang, Fei; R Elezgarai, Saioa; Bravo, Susana; Vázquez-Fernández, Ester; Rosa, Isaac; Eraña, Hasier; Gil, David; Veiga, Sonia; Vidal, Enric; Erickson-Beltran, Melissa L; Guitián, Esteban; Silva, Christopher J; Nonno, Romolo; Ma, Jiyan; Castilla, Joaquín; R Requena, Jesús

2018-01-01

Very solid evidence suggests that the core of full length PrPSc is a 4-rung β-solenoid, and that individual PrPSc subunits stack to form amyloid fibers. We recently used limited proteolysis to map the β-strands and connecting loops that make up the PrPSc solenoid. Using high resolution SDS-PAGE followed by epitope analysis, and mass spectrometry, we identified positions ~116/118, 133-134, 141, 152-153, 162, 169 and 179 (murine numbering) as Proteinase K (PK) cleavage sites in PrPSc. Such sites likely define loops and/or borders of β-strands, helping us to predict the threading of the β-solenoid. We have now extended this approach to recombinant PrPSc (recPrPSc). The term recPrPSc refers to bona fide recombinant prions prepared by PMCA, exhibiting infectivity with attack rates of ~100%. Limited proteolysis of mouse and bank vole recPrPSc species yielded N-terminally truncated PK-resistant fragments similar to those seen in brain-derived PrPSc, albeit with varying relative yields. Along with these fragments, doubly N- and C-terminally truncated fragments, in particular ~89/97-152, were detected in some recPrPSc preparations; similar fragments are characteristic of atypical strains of brain-derived PrPSc. Our results suggest a shared architecture of recPrPSc and brain PrPSc prions. The observed differences, in particular the distinct yields of specific PK-resistant fragments, are likely due to differences in threading which result in the specific biochemical characteristics of recPrPSc. Furthermore, recombinant PrPSc offers exciting opportunities for structural studies unachievable with brain-derived PrPSc.

A comparison of regression methods for model selection in individual-based landscape genetic analysis

Treesearch

Andrew J. Shirk; Erin L. Landguth; Samuel A. Cushman

2017-01-01

Anthropogenic migration barriers fragment many populations and limit the ability of species to respond to climate-induced biome shifts. Conservation actions designed to conserve habitat connectivity and mitigate barriers are needed to unite fragmented populations into larger, more viable metapopulations, and to allow species to track their climate envelope over time....

Twitter in the Higher Education Classroom: Known Fragmentations and Needed Frameworks

ERIC Educational Resources Information Center

Knowlton, Dave S.; Nygard, Shanda

2016-01-01

This article reviews early literature (that is, up to mid-year 2013) about Twitter's use in the higher education classroom. Fragmentations are highlighted as a way of showing that the literature does not cohesively claim advantages or disadvantages of using Twitter as a teaching and learning tool. The article also points to the limitations of the…

Effects of a one year physical activity program on serum C Terminal Agrin Fragment (CAF) concentrations among mobility limited older adults

USDA-ARS?s Scientific Manuscript database

OBJECTIVES: C terminal Agrin Fragment (CAF) has been proposed as a potential circulating biomarker for predicting changes in physical function among older adults. To determine the effect of a one year PA intervention on changes in CAF concentrations and to evaluate baseline and longitudinal associat...

An ab initio molecular dynamics study of S0 ketene fragmentation

NASA Astrophysics Data System (ADS)

Forsythe, Kelsey M.; Gray, Stephen K.; Klippenstein, Stephen J.; Hall, Gregory E.

2001-08-01

The dynamical origins of product state distributions in the unimolecular dissociation of S0 ketene, CH2CO (X˜ 1A1)→CH2(ã1A1)+CO, are studied with ab initio molecular dynamics. We focus on rotational distributions associated with ground vibrational state fragments. Trajectories are integrated between an inner, variational transition state (TS) and separated fragments in both the dissociative and associative directions. The average rotational energy in both CO and CH2 fragments decreases during the motion from the TS to separated fragments. However, the CO distribution remains slightly hotter than phase space theory (PST) predictions, whereas that for CH2 ends up significantly colder than PST, in good agreement with experiment. Our calculations do not, however, reproduce the experimentally observed correlations between CH2 and CO rotational states, in which the simultaneous formation of low rotational levels of each fragment is suppressed relative to PST. A limited search for nonstatistical behavior in the strong interaction region also fails to explain this discrepancy.

Dissecting fragment-based lead discovery at the von Hippel-Lindau protein:hypoxia inducible factor 1α protein-protein interface.

PubMed

Van Molle, Inge; Thomann, Andreas; Buckley, Dennis L; So, Ernest C; Lang, Steffen; Crews, Craig M; Ciulli, Alessio

2012-10-26

Fragment screening is widely used to identify attractive starting points for drug design. However, its potential and limitations to assess the tractability of often challenging protein:protein interfaces have been underexplored. Here, we address this question by means of a systematic deconstruction of lead-like inhibitors of the pVHL:HIF-1α interaction into their component fragments. Using biophysical techniques commonly employed for screening, we could only detect binding of fragments that violate the Rule of Three, are more complex than those typically screened against classical druggable targets, and occupy two adjacent binding subsites at the interface rather than just one. Analyses based on ligand and group lipophilicity efficiency of anchored fragments were applied to dissect the individual subsites and probe for binding hot spots. The implications of our findings for targeting protein interfaces by fragment-based approaches are discussed. Copyright © 2012 Elsevier Ltd. All rights reserved.

Object view in spatial system dynamics: a grassland farming example

PubMed Central

Neuwirth, Christian; Hofer, Barbara; Schaumberger, Andreas

2016-01-01

Abstract Spatial system dynamics (SSD) models are typically implemented by linking stock variables to raster grids while the use of object representations of human artefacts such as buildings or ownership has been limited. This limitation is addressed by this article, which demonstrates the use of object representations in SSD. The objects are parcels of land that are attributed to grassland farms. The model simulates structural change in agriculture, i.e., change in the size of farms. The aim of the model is to reveal relations between structural change, farmland fragmentation and variable farmland quality. Results show that fragmented farms tend to become consolidated by structural change, whereas consolidated initial conditions result in a significant increase of fragmentation. Consolidation is reinforced by a dynamic land market and high transportation costs. The example demonstrates the capabilities of the object-based approach for integrating object geometries (parcel shapes) and relations between objects (distances between parcels) dynamically in SSD. PMID:28190972

Tropical forest fragmentation affects floral visitors but not the structure of individual-based palm-pollinator networks.

PubMed

Dáttilo, Wesley; Aguirre, Armando; Quesada, Mauricio; Dirzo, Rodolfo

2015-01-01

Despite increasing knowledge about the effects of habitat loss on pollinators in natural landscapes, information is very limited regarding the underlying mechanisms of forest fragmentation affecting plant-pollinator interactions in such landscapes. Here, we used a network approach to describe the effects of forest fragmentation on the patterns of interactions involving the understory dominant palm Astrocaryum mexicanum (Arecaceae) and its floral visitors (including both effective and non-effective pollinators) at the individual level in a Mexican tropical rainforest landscape. Specifically, we asked: (i) Does fragment size affect the structure of individual-based plant-pollinator networks? (ii) Does the core of highly interacting visitor species change along the fragmentation size gradient? (iii) Does forest fragment size influence the abundance of effective pollinators of A. mexicanum? We found that fragment size did not affect the topological structure of the individual-based palm-pollinator network. Furthermore, while the composition of peripheral non-effective pollinators changed depending on fragment size, effective core generalist species of pollinators remained stable. We also observed that both abundance and variance of effective pollinators of male and female flowers of A. mexicanum increased with forest fragment size. These findings indicate that the presence of effective pollinators in the core of all forest fragments could keep the network structure stable along the gradient of forest fragmentation. In addition, pollination of A. mexicanum could be more effective in larger fragments, since the greater abundance of pollinators in these fragments may increase the amount of pollen and diversity of pollen donors between flowers of individual plants. Given the prevalence of fragmentation in tropical ecosystems, our results indicate that the current patterns of land use will have consequences on the underlying mechanisms of pollination in remnant forests.

Time-aware service-classified spectrum defragmentation algorithm for flex-grid optical networks

NASA Astrophysics Data System (ADS)

Qiu, Yang; Xu, Jing

2018-01-01

By employing sophisticated routing and spectrum assignment (RSA) algorithms together with a finer spectrum granularity (namely frequency slot) in resource allocation procedures, flex-grid optical networks can accommodate diverse kinds of services with high spectrum-allocation flexibility and resource-utilization efficiency. However, the continuity and the contiguity constraints in spectrum allocation procedures may always induce some isolated, small-sized, and unoccupied spectral blocks (known as spectrum fragments) in flex-grid optical networks. Although these spectrum fragments are left unoccupied, they can hardly be utilized by the subsequent service requests directly because of their spectral characteristics and the constraints in spectrum allocation. In this way, the existence of spectrum fragments may exhaust the available spectrum resources for a coming service request and thus worsens the networking performance. Therefore, many reactive defragmentation algorithms have been proposed to handle the fragmented spectrum resources via re-optimizing the routing paths and the spectrum resources for the existing services. But the routing-path and the spectrum-resource re-optimization in reactive defragmentation algorithms may possibly disrupt the traffic of the existing services and require extra components. By comparison, some proactive defragmentation algorithms (e.g. fragmentation-aware algorithms) were proposed to suppress spectrum fragments from their generation instead of handling the fragmented spectrum resources. Although these proactive defragmentation algorithms induced no traffic disruption and required no extra components, they always left the generated spectrum fragments unhandled, which greatly affected their efficiency in spectrum defragmentation. In this paper, by comprehensively considering the characteristics of both the reactive and the proactive defragmentation algorithms, we proposed a time-aware service-classified (TASC) spectrum defragmentation algorithm, which simultaneously employed proactive and reactive mechanisms in suppressing spectrum fragments with the awareness of services' types and their duration times. By dividing the spectrum resources into several flexible groups according to services' types and limiting both the spectrum allocation and the spectrum re-tuning for a certain service inside one specific spectrum group according to its type, the proposed TASC defragmentation algorithm cannot only suppress spectrum fragments from generation inside each spectrum group, but also handle the fragments generated between two adjacent groups. In this way, the proposed TASC algorithm gains higher efficiency in suppressing spectrum fragments than both the reactive and the proactive defragmentation algorithms. Additionally, as the generation of spectrum fragments is retrained between spectrum groups and the defragmentation procedure is limited inside each spectrum group, the induced traffic disruption for the existing services can be possibly reduced. Besides, the proposed TASC defragmentation algorithm always re-tunes the spectrum resources of the service with the maximum duration time first in spectrum defragmentation procedure, which can further reduce spectrum fragments because of the fact that the services with longer duration times always have higher possibility in inducing spectrum fragments than the services with shorter duration times. The simulation results show that the proposed TASC defragmentation algorithm can significantly reduce the number of the generated spectrum fragments while improving the service blocking performance.

Preparation of next-generation sequencing libraries using Nextera™ technology: simultaneous DNA fragmentation and adaptor tagging by in vitro transposition.

PubMed

Caruccio, Nicholas

2011-01-01

DNA library preparation is a common entry point and bottleneck for next-generation sequencing. Current methods generally consist of distinct steps that often involve significant sample loss and hands-on time: DNA fragmentation, end-polishing, and adaptor-ligation. In vitro transposition with Nextera™ Transposomes simultaneously fragments and covalently tags the target DNA, thereby combining these three distinct steps into a single reaction. Platform-specific sequencing adaptors can be added, and the sample can be enriched and bar-coded using limited-cycle PCR to prepare di-tagged DNA fragment libraries. Nextera technology offers a streamlined, efficient, and high-throughput method for generating bar-coded libraries compatible with multiple next-generation sequencing platforms.

Fragment-based drug discovery as alternative strategy to the drug development for neglected diseases.

PubMed

Mello, Juliana da Fonseca Rezende E; Gomes, Renan Augusto; Vital-Fujii, Drielli Gomes; Ferreira, Glaucio Monteiro; Trossini, Gustavo Henrique Goulart

2017-12-01

Neglected diseases (NDs) affect large populations and almost whole continents, representing 12% of the global health burden. In contrast, the treatment available today is limited and sometimes ineffective. Under this scenery, the Fragment-Based Drug Discovery emerged as one of the most promising alternatives to the traditional methods of drug development. This method allows achieving new lead compounds with smaller size of fragment libraries. Even with the wide Fragment-Based Drug Discovery success resulting in new effective therapeutic agents against different diseases, until this moment few studies have been applied this approach for NDs area. In this article, we discuss the basic Fragment-Based Drug Discovery process, brief successful ideas of general applications and show a landscape of its use in NDs, encouraging the implementation of this strategy as an interesting way to optimize the development of new drugs to NDs. © 2017 John Wiley & Sons A/S.

The coordination between mechanical and chemical subsystems initiates locomotion of Physarum plasmodial fragments

NASA Astrophysics Data System (ADS)

Zhang, Shun; Guy, Robert; Del Alamo, Juan Carlos

2017-11-01

Physarum polycephalum is a multinucleated slime mold whose endoplasm flows periodically driven by the contraction of its ectoplasm, a dense shell of F-actin cross-linked by myosin molecular motors and attached to the cell membrane. We find that physarum fragments smaller than 100 microns remain round and stay in place. However, larger fragments break symmetry leading to sustained forward locomotion, in process that is reminiscent of an interfacial instability that seems to settle around two different limit cycles (traveling waves and standing waves). We use both theory and experiments to study how coordination emerges between the different mechanical and chemical subsystems of the fragment to initiate locomotion. The role of many involved factors, such as fragment size, substratum adhesiveness, rheological properties, actin polymerization and traction stresses are investigated, and we find they agree well with our predictive model.

Using fragmentation trees and mass spectral trees for identifying unknown compounds in metabolomics.

PubMed

Vaniya, Arpana; Fiehn, Oliver

2015-06-01

Identification of unknown metabolites is the bottleneck in advancing metabolomics, leaving interpretation of metabolomics results ambiguous. The chemical diversity of metabolism is vast, making structure identification arduous and time consuming. Currently, comprehensive analysis of mass spectra in metabolomics is limited to library matching, but tandem mass spectral libraries are small compared to the large number of compounds found in the biosphere, including xenobiotics. Resolving this bottleneck requires richer data acquisition and better computational tools. Multi-stage mass spectrometry (MSn) trees show promise to aid in this regard. Fragmentation trees explore the fragmentation process, generate fragmentation rules and aid in sub-structure identification, while mass spectral trees delineate the dependencies in multi-stage MS of collision-induced dissociations. This review covers advancements over the past 10 years as a tool for metabolite identification, including algorithms, software and databases used to build and to implement fragmentation trees and mass spectral annotations.

Two-dimensional finite-element analyses of simulated rotor-fragment impacts against rings and beams compared with experiments

NASA Technical Reports Server (NTRS)

Stagliano, T. R.; Witmer, E. A.; Rodal, J. J. A.

1979-01-01

Finite element modeling alternatives as well as the utility and limitations of the two dimensional structural response computer code CIVM-JET 4B for predicting the transient, large deflection, elastic plastic, structural responses of two dimensional beam and/or ring structures which are subjected to rigid fragment impact were investigated. The applicability of the CIVM-JET 4B analysis and code for the prediction of steel containment ring response to impact by complex deformable fragments from a trihub burst of a T58 turbine rotor was studied. Dimensional analysis considerations were used in a parametric examination of data from engine rotor burst containment experiments and data from sphere beam impact experiments. The use of the CIVM-JET 4B computer code for making parametric structural response studies on both fragment-containment structure and fragment-deflector structure was illustrated. Modifications to the analysis/computation procedure were developed to alleviate restrictions.

Postfragmentation density function for bacterial aggregates in laminar flow

PubMed Central

Byrne, Erin; Dzul, Steve; Solomon, Michael; Younger, John

2014-01-01

The postfragmentation probability density of daughter flocs is one of the least well-understood aspects of modeling flocculation. We use three-dimensional positional data of Klebsiella pneumoniae bacterial flocs in suspension and the knowledge of hydrodynamic properties of a laminar flow field to construct a probability density function of floc volumes after a fragmentation event. We provide computational results which predict that the primary fragmentation mechanism for large flocs is erosion. The postfragmentation probability density function has a strong dependence on the size of the original floc and indicates that most fragmentation events result in clumps of one to three bacteria eroding from the original floc. We also provide numerical evidence that exhaustive fragmentation yields a limiting density inconsistent with the log-normal density predicted in the literature, most likely due to the heterogeneous nature of K. pneumoniae flocs. To support our conclusions, artificial flocs were generated and display similar postfragmentation density and exhaustive fragmentation. PMID:21599205

Loss of genetic connectivity and diversity in urban microreserves in a southern California endemic Jerusalem cricket (Orthoptera: Stenopelmatidae: Stenopelmatus n. sp. "santa monica")

USGS Publications Warehouse

Vandergast, A.G.; Lewallen, E.A.; Deas, J.; Bohonak, A.J.; Weissman, D.B.; Fisher, R.N.

2009-01-01

Microreserves may be useful in protecting native arthropod diversity in urbanized landscapes. However, species that do not disperse through the urban matrix may eventually be lost from these fragments. Population extinctions may be precipitated by an increase in genetic differentiation among fragments and loss of genetic diversity within fragments, and these effects should become stronger with time. We analyzed population genetic structure in the dispersal limited Jerusalem cricket Stenopelmatus n. sp. "santa monica" in the Santa Monica Mountains and Simi Hills north of Los Angeles, California (CA), to determine the impacts of fragmentation over the past 70 years. MtDNA divergence was greater among urban fragments than within contiguous habitat and was positively correlated with fragment age. MtDNA genetic diversity within fragments increased with fragment size and decreased with fragment age. Genetic divergence across 38 anonymous nuclear Inter-Simple Sequence Repeat (ISSR) loci was influenced by the presence of major highways and highway age, but there was no effect of additional urban fragmentation. ISSR diversity was not correlated with fragment size or age. Differing results between markers may be due to male-biased dispersal, or different effective population sizes, sorting rates, or mutation rates among sampled genes. Results suggest that genetic connectivity among populations has been disrupted by highways and urban development, prior to declines in local population sizes. We emphasize that genetic connectivity can rapidly erode in fragmented landscapes and that flightless arthropods can serve as sensitive indicators for these effects. ?? Springer Science+Business Media B.V. 2008.

FRAGSION: ultra-fast protein fragment library generation by IOHMM sampling.

PubMed

Bhattacharya, Debswapna; Adhikari, Badri; Li, Jilong; Cheng, Jianlin

2016-07-01

Speed, accuracy and robustness of building protein fragment library have important implications in de novo protein structure prediction since fragment-based methods are one of the most successful approaches in template-free modeling (FM). Majority of the existing fragment detection methods rely on database-driven search strategies to identify candidate fragments, which are inherently time-consuming and often hinder the possibility to locate longer fragments due to the limited sizes of databases. Also, it is difficult to alleviate the effect of noisy sequence-based predicted features such as secondary structures on the quality of fragment. Here, we present FRAGSION, a database-free method to efficiently generate protein fragment library by sampling from an Input-Output Hidden Markov Model. FRAGSION offers some unique features compared to existing approaches in that it (i) is lightning-fast, consuming only few seconds of CPU time to generate fragment library for a protein of typical length (300 residues); (ii) can generate dynamic-size fragments of any length (even for the whole protein sequence) and (iii) offers ways to handle noise in predicted secondary structure during fragment sampling. On a FM dataset from the most recent Critical Assessment of Structure Prediction, we demonstrate that FGRAGSION provides advantages over the state-of-the-art fragment picking protocol of ROSETTA suite by speeding up computation by several orders of magnitude while achieving comparable performance in fragment quality. Source code and executable versions of FRAGSION for Linux and MacOS is freely available to non-commercial users at http://sysbio.rnet.missouri.edu/FRAGSION/ It is bundled with a manual and example data. chengji@missouri.edu Supplementary data are available at Bioinformatics online. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Contrasting patterns of nest survival and postfledging survival in ovenbirds and Acadian flycatchers in Missouri forest fragments

Treesearch

Julianna M. A. Jenkins; Frank R. Thompson; John Faaborg

2016-01-01

We can improve our ability to assess population viability and forecast population growth under different scenarios by understanding factors that limit population parameters in each stage of the annual cycle. Postfledging mortality rates may be as variable as nest survival across regions and fragmentation gradients, although factors that negatively impact nest survival...

Palaeoproteomics for human evolution studies

NASA Astrophysics Data System (ADS)

Welker, Frido

2018-06-01

The commonplace sequencing of Neanderthal, Denisovan and ancient modern human DNA continues to revolutionize our understanding of hominin phylogeny and interaction(s). The challenge with older fossils is that the progressive fragmentation of DNA even under optimal conditions, a function of time and temperature, results in ever shorter fragments of DNA. This process continues until no DNA can be sequenced or reliably aligned. Ancient proteins ultimately suffer a similar fate, but are a potential alternative source of biomolecular sequence data to investigate hominin phylogeny given their slower rate of fragmentation. In addition, ancient proteins have been proposed to potentially provide insights into in vivo biological processes and can be used to provide additional ecological information through large scale ZooMS (Zooarchaeology by Mass Spectrometry) screening of unidentifiable bone fragments. However, as initially with ancient DNA, most ancient protein research has focused on Late Pleistocene or Holocene samples from Europe. In addition, only a limited number of studies on hominin remains have been published. Here, an updated review on ancient protein analysis in human evolutionary contexts is given, including the identification of specific knowledge gaps and existing analytical limits, as well as potential avenues to overcome these.

Dispersal Limitations on Fish Community Recovery Following Long-term Water Quality Remediation

DOE PAGES

McManamay, Ryan A.; Jett, Robert T.; Ryon, Michael G.; ...

2016-02-22

Holistic restoration approaches, such as water quality remediation, are likely to meet conservation objectives because they are typically implemented at watershed scales, as opposed to individual stream reaches. However, habitat fragmentation may impose constraints on the ecological effectiveness of holistic restoration strategies by limiting colonization following remediation. We questioned the importance of dispersal limitations to fish community recovery following long-term water quality remediation and species reintroductions across the White Oak Creek (WOC) watershed near Oak Ridge, Tennessee (USA). Long-term (26 years) responses in fish species richness and biomass to water quality remediation were evaluated in light of habitat fragmentation andmore » population isolation from instream barriers, which varied in their passage potential. In addition, ordination techniques were used to determine the relative importance of habitat connectivity and water quality, in explaining variation fish communities relative to environmental fluctuations, i.e. streamflow. Ecological recovery (changes in richness) at each site was negatively related to barrier index, a measure of community isolation by barriers relative to stream distance. Following species reintroductions, dispersal by fish species was consistently in the downstream direction and upstream passage above barriers was non-existent. The importance of barrier index in explaining variation in fish communities was stronger during higher flow conditions, but decreased over time an indication of increasing community stability and loss of seasonal migrants. Compared to habitat fragmentation, existing water quality concerns (i.e., outfalls, point source discharges) were unrelated to ecological recovery, but explained relatively high variation in community dynamics. Our results suggest that habitat fragmentation limited the ecological effectiveness of intensive water quality remediation efforts and fish reintroduction efforts by impeding recolonization at isolated stream reaches.« less

Dispersal Limitations on Fish Community Recovery Following Long-term Water Quality Remediation

DOE Office of Scientific and Technical Information (OSTI.GOV)

McManamay, Ryan A.; Jett, Robert T.; Ryon, Michael G.

Holistic restoration approaches, such as water quality remediation, are likely to meet conservation objectives because they are typically implemented at watershed scales, as opposed to individual stream reaches. However, habitat fragmentation may impose constraints on the ecological effectiveness of holistic restoration strategies by limiting colonization following remediation. We questioned the importance of dispersal limitations to fish community recovery following long-term water quality remediation and species reintroductions across the White Oak Creek (WOC) watershed near Oak Ridge, Tennessee (USA). Long-term (26 years) responses in fish species richness and biomass to water quality remediation were evaluated in light of habitat fragmentation andmore » population isolation from instream barriers, which varied in their passage potential. In addition, ordination techniques were used to determine the relative importance of habitat connectivity and water quality, in explaining variation fish communities relative to environmental fluctuations, i.e. streamflow. Ecological recovery (changes in richness) at each site was negatively related to barrier index, a measure of community isolation by barriers relative to stream distance. Following species reintroductions, dispersal by fish species was consistently in the downstream direction and upstream passage above barriers was non-existent. The importance of barrier index in explaining variation in fish communities was stronger during higher flow conditions, but decreased over time an indication of increasing community stability and loss of seasonal migrants. Compared to habitat fragmentation, existing water quality concerns (i.e., outfalls, point source discharges) were unrelated to ecological recovery, but explained relatively high variation in community dynamics. Our results suggest that habitat fragmentation limited the ecological effectiveness of intensive water quality remediation efforts and fish reintroduction efforts by impeding recolonization at isolated stream reaches.« less

[Avulsion fracture of the calcaneal tuberosity in athletes].

PubMed

Glanzmann, M; Veréb, L; Habegger, R

2005-04-01

Avulsion fractures of the calcaneal tuberosity are rare injuries. Several surgical treatment options have been described. The size of the calcaneal fragment is the limiting factor in choosing the method for restoration. Finding the right type of fixation modality remains challenging in this rare kind of injury. In the case presented one Mitek Super Anchor was used to reattach the small fragment of the tuberosity to the calcaneus. Stable fixation with bony reunion and excellent functional outcome were achieved by this technique within 10 weeks. Therefore, we recommend the use of an anchor system for the treatment of small fragment calcaneal avulsion fractures.

Evacuation areas for transportation accidents involving propellant tank pressure bursts

NASA Technical Reports Server (NTRS)

Siewert, R. D.

1972-01-01

Evacuation areas are defined for those transportation accidents where volatile chemical propellant tanks are exposed to fire in the wreckage and eventually explode with consequent risks from fragments in surrounding populated areas. An evacuation area with a minimum radius of 600 m (2000 ft) is recommended to limit the statistical probability of fatality to one in 100 such accidents. The result was made possible by the derivation of a distribution function of distances reached by fragments from bursting chemical car tanks. Data concerning fragments was obtained from reports or tank car pressure bursts between 1958 and 1971.

Plant reproductive susceptibility to habitat fragmentation: review and synthesis through a meta-analysis.

PubMed

Aguilar, Ramiro; Ashworth, Lorena; Galetto, Leonardo; Aizen, Marcelo Adrián

2006-08-01

The loss and fragmentation of natural habitats by human activities are pervasive phenomena in terrestrial ecosystems across the Earth and the main driving forces behind current biodiversity loss. Animal-mediated pollination is a key process for the sexual reproduction of most extant flowering plants, and the one most consistently studied in the context of habitat fragmentation. By means of a meta-analysis we quantitatively reviewed the results from independent fragmentation studies throughout the last two decades, with the aim of testing whether pollination and reproduction of plant species may be differentially susceptible to habitat fragmentation depending on certain reproductive traits that typify the relationship with and the degree of dependence on their pollinators. We found an overall large and negative effect of fragmentation on pollination and on plant reproduction. The compatibility system of plants, which reflects the degree of dependence on pollinator mutualism, was the only reproductive trait that explained the differences among the species' effect sizes. Furthermore, a highly significant correlation between the effect sizes of fragmentation on pollination and reproductive success suggests that the most proximate cause of reproductive impairment in fragmented habitats may be pollination limitation. We discuss the conservation implications of these findings and give some suggestions for future research into this area.

Raising an Antibody Specific to Breast Cancer Subpopulations Using Phage Display on Tissue Sections.

PubMed

Larsen, Simon Asbjørn; Meldgaard, Theresa; Fridriksdottir, Agla Jael Rubner; Lykkemark, Simon; Poulsen, Pi Camilla; Overgaard, Laura Falkensteen; Petersen, Helene Bundgaard; Petersen, Ole William; Kristensen, Peter

2016-01-01

Primary tumors display a great level of intra-tumor heterogeneity in breast cancer. The current lack of prognostic and predictive biomarkers limits accurate stratification and the ability to predict response to therapy. The aim of the present study was to select recombinant antibody fragments specific against breast cancer subpopulations, aiding the discovery of novel biomarkers. Recombinant antibody fragments were selected by phage display. A novel shadowstick technology enabled the direct selection using tissue sections of antibody fragments specific against small subpopulations of breast cancer cells. Selections were performed against a subpopulation of breast cancer cells expressing CD271+, as these previously have been indicated to be potential breast cancer stem cells. The selected antibody fragments were screened by phage ELISA on both breast cancer and myoepithelial cells. The antibody fragments were validated and evaluated by immunohistochemistry experiments. Our study revealed an antibody fragment, LH8, specific for breast cancer cells. Immunohistochemistry results indicate that this particular antibody fragment binds an antigen that exhibits differential expression in different breast cancer subpopulations. Further studies characterizing this antibody fragment, the subpopulation it binds and the cognate antigen may unearth novel biomarkers of clinical relevance. Copyright© 2016, International Institute of Anticancer Research (Dr. John G. Delinasios), All rights reserved.

Fusion of small unilamellar vesicles induced by bovine serum albumin fragments.

PubMed

Garcia, L A; Schenkman, S; Araujo, P S; Chaimovich, H

1983-07-01

The limited pepsin proteolysis products of bovine serum albumin, fragment A (residues 307-586) and fragment B (residues 1-306), induced the fusion of small unilamellar vesicles of egg phosphatidyl choline at concentrations near 5 microM. Fusion was demonstrated and analyzed on the basis of: a) time-dependent changes in absorbance; b) dilution of the fluorescent label 2-(10-(1-pyrene)decanoyl) phosphatidyl choline, incorporated into a small percentage of the vesicles, as measured by the decrease in the excimer to monomer (E/M) ratio; c) increase of the average hydrodynamic radius of the liposomes, estimated by Sepharose 4B filtration, and d) the strict inverse relationship between the size of the liposomes and their E/M ratios. Albumin fragment B, like albumin, induced the formation of large aggregates in which rapid cooperative fusion produced vesicles having a large hydrodynamic radius. Fragment A did not produce large aggregates and the initial fusion products exhibited a hydrodynamic radius. Fragment A did not produce large aggregates and the initial fusion products exhibited a hydrodynamic radius smaller than those obtained with fragment B. Albumin and fragments A and B are fusogenic only at pH below 4.0. These data discussed in terms of a general model for a signal-dependent protein-induced membrane fusion.

A New Device for Percutaneous Elevation of the Depressed Fractures of Tibial Condyles

PubMed Central

Ravindranath, V.S.; Kumar, Madhusudan; Murthy, G.V.S.

2012-01-01

Introduction: Monocondylar tibia plateau fractures with non-comminuted fragments can be treated using percutaneous screws. Currently indirect methods of reduction are used and thus the technique is limited to fragments with less than 5 mm depression. The first author has designed a device for direct elevation and reduction of the fragments thus potentially expanding the indications of percutaneous screws to fragments with >5mm depression Technical Note: A total of ten cases were treated by this method of percutaneous elevation of the depressed fractures of lateral condyles of the Tibia using this device. Device was inserted through a bony window on the anteromedial surface of tibia. The inner piston of the device in slowly hammered inside thus elevating the depressed fragment. Elevation of fragment could be achieved in all the cases. The fractures were fixed with cancellous screws applied percutaneously. There were no cases with loss of fixation or subsidence of the fragment. All cases achieved radiological union and have good knee function at follow up Conclusion: The new device is able to elevate unicondylar tibia plateau fragments with no subsidence or loss of fixation in our series. A longer follow up in a larger sample will be needed to establish the technique. PMID:27298860

Identification of linker regions and domain borders of the transcription activator protein NtrC from Escherichia coli by limited proteolysis, in-gel digestion, and mass spectrometry.

PubMed

Bantscheff, M; Weiss, V; Glocker, M O

1999-08-24

We have developed a mass spectrometry based method for the identification of linker regions and domain borders in multidomain proteins. This approach combines limited proteolysis and in-gel proteolytic digestions and was applied to the determination of linkers in the transcription factor NtrC from Escherichia coli. Limited proteolysis of NtrC with thermolysin and papain revealed that initial digestion yielded two major bands in SDS-PAGE that were identified by mass spectrometry as the R-domain and the still covalently linked OC-domains. Subsequent steps in limited proteolysis afforded further cleavage of the OC-fragment into the O- and the C-domain at accessible amino acid residues. Mass spectrometric identification of the tryptic/thermolytic peptides obtained after in-gel total proteolysis of the SDS-PAGE-separated domains determined the domain borders and showed that the protease accessible linker between R- and O-domain comprised amino acids Val-131 and Gln-132 within the "Q-linker" in agreement with papain and subtilisin digestion. The region between amino acid residues Thr-389 and Gln-396 marked the hitherto unknown linker sequence that connects the O- with the C-domain. High abundances of proline-, alanine-, serine-, and glutamic acid residues were found in this linker structure (PASE-linker) of related NtrC response regulator proteins. While R- and C-domains remained stable under the applied limited proteolysis conditions, the O-domain was further truncated yielding a core fragment that comprised the sequence from Ile-140 to Arg-320. ATPase activity was lost after separation of the R-domain from the OC-fragment. However, binding of OC- and C- fragments to specific DNA was observed by characteristic band-shifts in migration retardation assays, indicating intact tertiary structures of the C-domain. The outlined strategy proved to be highly efficient and afforded lead information of tertiary structural features necessary for protein design and engineering and for structure-function studies.

Structural insights into binding of inhibitors to soluble epoxide hydrolase gained by fragment screening and X-ray crystallography.

PubMed

Amano, Yasushi; Yamaguchi, Tomohiko; Tanabe, Eiki

2014-04-15

Soluble epoxide hydrolase (sEH) is a component of the arachidonic acid cascade and is a candidate target for therapies for hypertension or inflammation. Although many sEH inhibitors are available, their scaffolds are not structurally diverse, and knowledge of their specific interactions with sEH is limited. To obtain detailed structural information about protein-ligand interactions, we conducted fragment screening of sEH, analyzed the fragments using high-throughput X-ray crystallography, and determined 126 fragment-bound structures at high resolution. Aminothiazole and benzimidazole derivatives were identified as novel scaffolds that bind to the catalytic triad of sEH with good ligand efficiency. We further identified fragment hits that bound to subpockets of sEH called the short and long branches. The water molecule conserved in the structure plays an important role in binding to the long branch, whereas Asp496 and the main chain of Phe497 form hydrogen bonds with fragment hits in the short branch. Fragment hits and their crystal structures provide structural insights into ligand binding to sEH that will facilitate the discovery of novel and potent inhibitors of sEH. Copyright © 2014 Elsevier Ltd. All rights reserved.

A Ligand-observed Mass Spectrometry Approach Integrated into the Fragment Based Lead Discovery Pipeline

PubMed Central

Chen, Xin; Qin, Shanshan; Chen, Shuai; Li, Jinlong; Li, Lixin; Wang, Zhongling; Wang, Quan; Lin, Jianping; Yang, Cheng; Shui, Wenqing

2015-01-01

In fragment-based lead discovery (FBLD), a cascade combining multiple orthogonal technologies is required for reliable detection and characterization of fragment binding to the target. Given the limitations of the mainstream screening techniques, we presented a ligand-observed mass spectrometry approach to expand the toolkits and increase the flexibility of building a FBLD pipeline especially for tough targets. In this study, this approach was integrated into a FBLD program targeting the HCV RNA polymerase NS5B. Our ligand-observed mass spectrometry analysis resulted in the discovery of 10 hits from a 384-member fragment library through two independent screens of complex cocktails and a follow-up validation assay. Moreover, this MS-based approach enabled quantitative measurement of weak binding affinities of fragments which was in general consistent with SPR analysis. Five out of the ten hits were then successfully translated to X-ray structures of fragment-bound complexes to lay a foundation for structure-based inhibitor design. With distinctive strengths in terms of high capacity and speed, minimal method development, easy sample preparation, low material consumption and quantitative capability, this MS-based assay is anticipated to be a valuable addition to the repertoire of current fragment screening techniques. PMID:25666181

Morphometric analysis of chameleon fossil fragments from the Early Pliocene of South Africa: a new piece of the chamaeleonid history.

PubMed

Dollion, Alexis Y; Cornette, Raphaël; Tolley, Krystal A; Boistel, Renaud; Euriat, Adelaïde; Boller, Elodie; Fernandez, Vincent; Stynder, Deano; Herrel, Anthony

2015-02-01

The evolutionary history of chameleons has been predominantly studied through phylogenetic approaches as the fossil register of chameleons is limited and fragmented. The poor state of preservation of these fossils has moreover led to the origin of numerous nomen dubia, and the identification of many chameleon fossils remains uncertain. We here examine chameleon fossil fragments from the Early Pliocene Varswater formation, exposed at the locality of Langebaanweg "E" Quarry along the southwestern coast of South Africa. Our aim was to explore whether these fossil fragments could be assigned to extant genera. To do so, we used geometric morphometric approaches based on microtomographic imaging of extant chameleons as well as the fossil fragments themselves. Our study suggests that the fossils from this deposit most likely represent at least two different forms that may belong to different genera. Most fragments are phenotypically dissimilar from the South African endemic genus Bradypodion and are more similar to other chameleon genera such as Trioceros or Kinyongia. However, close phenetic similarities between some of the fragments and the Seychelles endemic Archaius or the Madagascan genus Furcifer suggest that some of these fragments may not contain enough genus-specific information to allow correct identification. Other fragments such as the parietal fragments appear to contain more genus-specific information, however. Although our data suggest that the fossil diversity of chameleons in South Africa was potentially greater than it is today, this remains to be verified based on other and more complete fragments.

Morphometric analysis of chameleon fossil fragments from the Early Pliocene of South Africa: a new piece of the chamaeleonid history

NASA Astrophysics Data System (ADS)

Dollion, Alexis Y.; Cornette, Raphaël; Tolley, Krystal A.; Boistel, Renaud; Euriat, Adelaïde; Boller, Elodie; Fernandez, Vincent; Stynder, Deano; Herrel, Anthony

2015-02-01

The evolutionary history of chameleons has been predominantly studied through phylogenetic approaches as the fossil register of chameleons is limited and fragmented. The poor state of preservation of these fossils has moreover led to the origin of numerous nomen dubia, and the identification of many chameleon fossils remains uncertain. We here examine chameleon fossil fragments from the Early Pliocene Varswater formation, exposed at the locality of Langebaanweg "E" Quarry along the southwestern coast of South Africa. Our aim was to explore whether these fossil fragments could be assigned to extant genera. To do so, we used geometric morphometric approaches based on microtomographic imaging of extant chameleons as well as the fossil fragments themselves. Our study suggests that the fossils from this deposit most likely represent at least two different forms that may belong to different genera. Most fragments are phenotypically dissimilar from the South African endemic genus Bradypodion and are more similar to other chameleon genera such as Trioceros or Kinyongia. However, close phenetic similarities between some of the fragments and the Seychelles endemic Archaius or the Madagascan genus Furcifer suggest that some of these fragments may not contain enough genus-specific information to allow correct identification. Other fragments such as the parietal fragments appear to contain more genus-specific information, however. Although our data suggest that the fossil diversity of chameleons in South Africa was potentially greater than it is today, this remains to be verified based on other and more complete fragments.

Luminous Efficiency Estimates of Meteors. II. Application to Canadian Automated Meteor Observatory Meteor Events

NASA Astrophysics Data System (ADS)

Subasinghe, Dilini; Campbell-Brown, Margaret

2018-02-01

Luminous efficiency is a necessary parameter for determining meteoroid mass from optical emission. Despite this importance, it is very poorly known, with previous results varying by up to two orders of magnitude for a given speed. We present the most recent study of luminous efficiency values determined with modern high-resolution instruments, by directly comparing dynamic and photometric meteoroid masses. Fifteen non-fragmenting meteoroids were used, with a further five clearly fragmenting events for comparison. Twelve of the fifteen non-fragmenting meteoroids had luminous efficiencies less than 1%, while the fragmenting meteoroids had upper limits of a few tens of per cent. No clear trend with speed was seen, but there was a weak negative trend of luminous efficiency on meteoroid mass, implying that smaller meteoroids radiate more efficiently.

Lava bubble-wall fragments formed by submarine hydrovolcanic explosions on Lo'ihi Seamount and Kilauea Volcano

USGS Publications Warehouse

Clague, D.A.; Davis, A.S.; Bischoff, J.L.; Dixon, J.E.; Geyer, R.

2000-01-01

Glassy bubble-wall fragments, morphologically similar to littoral limu o Pele, have been found in volcanic sands erupted on Lo'ihi Seamount and along the submarine east rift zone of Kilauea Volcano. The limu o Pele fragments are undegassed with respect to H2O and S and formed by mild steam explosions. Angular glass sand fragments apparently form at similar, and greater, depths by cooling-contraction granulation. The limu o Pele fragments from Lo'ihi Seamount are dominantly tholeiitic basalt containing 6.25-7.25% MgO. None of the limu o Pele samples from Lo'ihi Seamount contains less than 5.57% MgO, suggesting that higher viscosity magmas do not form lava bubbles. The dissolved CO2 and H2O contents of 7 of the limu o Pele fragments indicate eruption at 1200??300 m depth (120??30 bar). These pressures exceed that generally thought to limit steam explosions. We conclude that hydrovolcanic eruptions are possible, with appropriate pre-mixing conditions, at pressures as great as 120 bar.

Simulated effects of climate change, fragmentation, and inter-specific competition on tree species migration in northern Wisconsin, USA

Treesearch

Robert M. Scheller; David J. Mladenoff

2008-01-01

The reproductive success, growth, and mortality rates of tree species in the northern United States will be differentially affected by projected climate change over the next century. As a consequence, the spatial distributions of tree species will expand or contract at differential rates. In addition, human fragmentation of the landscape may limit effective seed...

Subjectivity in Design Education: The Perception of the City through Personal Maps

ERIC Educational Resources Information Center

Yilmaz, Ebru

2016-01-01

Our mental maps related to the cities are limited by our personal perception and fragmented in the process. There are many inner and outer effects that shape our mental maps, and as a result the fragmented whole refers to the total city image in our minds. To represent this image, an experimental study has been conducted with a group of students.…

Fragmentation and matrix contrast favor understory plants through negative cascading effects on a strong competitor palm.

PubMed

Hernández-Ruedas, Manuel A; Arroyo-Rodríguez, Víctor; Morante-Filho, José Carlos; Meave, Jorge A; Martínez-Ramos, Miguel

2018-05-04

Understanding the patterns and processes driving biodiversity maintenance in fragmented tropical forests is urgently needed for conservation planning, especially in species-rich forest reserves. Of particular concern are the effects that habitat modifications at the landscape scale may have on forest regeneration and ecosystem functioning: a topic that has received limited attention. Here, we assessed the effects of landscape structure (i.e., forest cover, open area matrices, forest fragmentation, and mean inter-patch isolation distance) on understory plant assemblages in the Los Tuxtlas Biosphere Reserve, Mexico. Previous studies suggest that the demographic burst of the strong competitor palm Astrocaryum mexicanum in the core area of this reserve limits plant recruitment and imperils biodiversity conservation within this protected area. Yet, the local and landscape predictors of this palm, and its impact on tree recruitment at a regional scale are unknown. Thus, we used structural equation modeling to assess the direct and cascading effects of landscape structure on stem and species density in the understory of 20 forest sites distributed across this biodiversity hotspot. Indirect paths included the effect of landscape structure on tree basal area (a proxy of local disturbance), and the effects of these variables on A. mexicanum. Density of A. mexicanum mainly increased with decreasing both fragmentation and open areas in the matrix (matrix contrast, hereafter), and such an increase in palm density negatively affected stem and species density in the understory. The negative direct effect of matrix contrast on stem density was overridden by the indirect positive effects (i.e., through negative cascading effects on A. mexicanum), resulting in a weak effect of matrix contrast on stem density. These findings suggest that dispersal limitation and negative edge effects in more fragmented landscapes dominated by open areas prevent the proliferation of this palm species, enhancing the diversity and abundance of understory trees. This "positive" news adds to an increasing line of evidence suggesting that fragmentation may have some positive effects on biodiversity, in this case by preventing the proliferation of species that can jeopardize biodiversity conservation within tropical reserves. © 2018 by the Ecological Society of America.

Native top-down mass spectrometry for the structural characterization of human hemoglobin

DOE PAGES

Zhang, Jiang; Malmirchegini, G. Reza; Clubb, Robert T.; ...

2015-06-09

Native mass spectrometry (MS) has become an invaluable tool for the characterization of proteins and non-covalent protein complexes under near physiological solution conditions. Here we report the structural characterization of human hemoglobin (Hb), a 64 kDa oxygen-transporting protein complex, by high resolution native top-down mass spectrometry using electrospray ionization (ESI) and a 15-Tesla Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. Native MS preserves the non-covalent interactions between the globin subunits, and electron capture dissociation (ECD) produces fragments directly from the intact Hb complex without dissociating the subunits. Using activated ion ECD, we observe the gradual unfolding process of themore » Hb complex in the gas phase. Without protein ion activation, the native Hb shows very limited ECD fragmentation from the N-termini, suggesting a tightly packed structure of the native complex and therefore low fragmentation efficiency. Precursor ion activation allows steady increase of N-terminal fragment ions, while the C-terminal fragments remain limited (38 c ions and 4 z ions on the α chain; 36 c ions and 2 z ions on the β chain). This ECD fragmentation pattern suggests that upon activation, the Hb complex starts to unfold from the N-termini of both subunits, whereas the C-terminal regions and therefore the potential regions involved in the subunit binding interactions remain intact. ECD-MS of the Hb dimer show similar fragmentation patterns as the Hb tetramer, providing further evidence for the hypothesized unfolding process of the Hb complex in the gas phase. Native top-down ECD-MS allows efficient probing of the Hb complex structure and the subunit binding interactions in the gas phase. Finally, it may provide a fast and effective means to probe the structure of novel protein complexes that are intractable to traditional structural characterization tools.« less

Gas-phase fragmentation of peptides by MALDI in-source decay with limited amide hydrogen (1H/2H) scrambling.

PubMed

Bache, Nicolai; Rand, Kasper D; Roepstorff, Peter; Jørgensen, Thomas J D

2008-08-15

To achieve a fundamental understanding of the function of proteins and protein complexes at the molecular level, it is crucial to obtain a detailed knowledge about their dynamic and structural properties. The kinetics of backbone amide hydrogen exchange is intimately linked to the structural dynamics of the protein, and in recent years, the monitoring of the isotopic exchange of these hydrogens by mass spectrometry has become a recognized method. At present, the resolution of this method is, however, limited and single-residue resolution is typically only obtained for a few residues in a protein. It would therefore be desirable if gas-phase fragmentation could be used to localize incorporated deuterons as this would ultimately lead to single-residue resolution. A central obstacle for this approach is, however, the occurrence of intramolecular migration of amide hydrogens upon activation of the gaseous protein (i.e., hydrogen scrambling). Here we investigate the occurrence of scrambling in selectively labeled peptides upon fragmentation by matrix-assisted laser desorption/ionization in-source decay (MALDI ISD). We have utilized peptides with a unique regioselective deuterium incorporation that allows us to accurately determine the extent of scrambling upon fragmentation. Our results show that the level of scrambling upon MALDI ISD is so low that the solution deuteration pattern is readily apparent in the gas-phase fragment ions. These results suggest that MALDI ISD may prove useful for hydrogen exchange studies of purified peptides and small proteins.

Tumor containing fragment number influences immunohistochemistry positive rate of HER2 in biopsy specimens of gastric cancer.

PubMed

Xu, Chen; Liu, Yalan; Ge, Xiaowen; Jiang, Dongxian; Zhang, Ying; Ji, Yuan; Hou, Jun; Huang, Jie; Su, Jieakesu; Zeng, Haiying; Qin, Jing; Hou, Yingyong

2017-05-26

HER2 assessment in biopsy specimens of gastric cancer (GC) is challenging because of the intratumoral heterogeneity. False negative results may be get because of limited biopsy material. The aim of this study is to explore how tumor-containing fragment number and biopsy specimen number affect HER2 immunohistochemistry (IHC) positive rate. Eight hundred and ninety biopsy specimens and 459 paired resected specimens were collected. IHC staining of HER2 was performed. HER2 IHC positive (scored 3+) rate was compared based on tumor-containing fragment number, biopsy specimen number, average size and tumor tissue proportion of tumor-containing fragments. The positive predictability of biopsy specimens to resected specimens was analyzed based on tumor fragment number. HER2 IHC positive rates were 2.0, 3.5, 7.0, 13.2, 17.1, and 15.9% when tumor fragment numbers were 1, 2, 3, 4, 5 and 6 respectively. The rate rose with the increase of tumor fragment number (P = 0.004). ROC curve analysis showed that biopsy specimens exhibited positive predictability when tumor fragment number reached 3, but showed better performance when the number was ≥4 (P < 0.05). After fragment number reached 4, no statistic differences were reached in either HER2 IHC positive rate or positive predictability with further increase of the number (P > 0.05). HER2 IHC positive rate was not associated with biopsy number (P = 0.127), average size of tumor fragments (P = 0.397), and tumor tissue proportion of tumor fragments (P = 0.825) directly. The number of tumor-containing fragments influences HER2 IHC positive (scored 3+) rate. Greater than or equal to 4 (≥4) tumor fragments give better results in the positive rate as well as positive predictability. We recommend the number of tumor containing fragments be described in the HER2 IHC pathology reports for clinical reference in endoscopic biopsy specimens of GC.

Earth's 2006 Encounter with Comet 73P/Schwassmann-Wachmann: Products of Nucleus Fragmentation Seen in Closeup

NASA Technical Reports Server (NTRS)

Sekanina, Zdenek

2007-01-01

The large numbers of nucleus fragments observed are a spectacular illustration of the process of cascading fragmentation in progress, a concept introduced to interpret the properties of the Kreutz system of sungrazers and comet D/1993 F2. The objective is to describe the fragmentation sequence and hierarchy of comet 73P, the nature of the fragmentation process and observed events, and the expected future evolution of this comet. The orbital arc populated by the fragments refers to an interval of 3.74 days in the perihelion time. This result suggests that they all could be products (but not necessarily first-generation fragments) of two 1995 events, in early September (involving an enormous outburst) and at the beginning of November. The interval of perihelion times is equivalent to a range of about 2.5 m/s in separation velocity or 0.00012 the Sun's attraction in nongravitational deceleration. Their combined effect suggests minor orbital momentum changes acquired during fragmentation and decelerations compatible with survival over two revolutions about the Sun. Fragment B is a likely first-generation product of one of the 1995 events. From the behavior of the primary fragment C, 73P is not a dying comet, even though fragment B and others were episodically breaking up into many pieces. Each episode began with the sudden appearance of a starlike nucleus condensation and a rapidly expanding outburst, followed by a development of jets, and a gradual tailward extension of the fading condensation, until the discrete masses embedded in it could be resolved. In April-May, this debris traveled first to the southwest, but models show their eventual motion toward the projected orbit. Fainter fragments were imaged over limited time, apparently because of their erratic activity (interspersed with periods of dormancy) rather than improptu disintegration. A dust trail joining the fragments and reminiscent of comet 141P/Machholz suggests that cascading fragmentation exerts itself profoundly over an extremely broad mass range of particulate debris.

Analysis of simple 2-D and 3-D metal structures subjected to fragment impact

NASA Technical Reports Server (NTRS)

Witmer, E. A.; Stagliano, T. R.; Spilker, R. L.; Rodal, J. J. A.

1977-01-01

Theoretical methods were developed for predicting the large-deflection elastic-plastic transient structural responses of metal containment or deflector (C/D) structures to cope with rotor burst fragment impact attack. For two-dimensional C/D structures both, finite element and finite difference analysis methods were employed to analyze structural response produced by either prescribed transient loads or fragment impact. For the latter category, two time-wise step-by-step analysis procedures were devised to predict the structural responses resulting from a succession of fragment impacts: the collision force method (CFM) which utilizes an approximate prediction of the force applied to the attacked structure during fragment impact, and the collision imparted velocity method (CIVM) in which the impact-induced velocity increment acquired by a region of the impacted structure near the impact point is computed. The merits and limitations of these approaches are discussed. For the analysis of 3-d responses of C/D structures, only the CIVM approach was investigated.

Postfragmentation density function for bacterial aggregates in laminar flow.

PubMed

Byrne, Erin; Dzul, Steve; Solomon, Michael; Younger, John; Bortz, David M

2011-04-01

The postfragmentation probability density of daughter flocs is one of the least well-understood aspects of modeling flocculation. We use three-dimensional positional data of Klebsiella pneumoniae bacterial flocs in suspension and the knowledge of hydrodynamic properties of a laminar flow field to construct a probability density function of floc volumes after a fragmentation event. We provide computational results which predict that the primary fragmentation mechanism for large flocs is erosion. The postfragmentation probability density function has a strong dependence on the size of the original floc and indicates that most fragmentation events result in clumps of one to three bacteria eroding from the original floc. We also provide numerical evidence that exhaustive fragmentation yields a limiting density inconsistent with the log-normal density predicted in the literature, most likely due to the heterogeneous nature of K. pneumoniae flocs. To support our conclusions, artificial flocs were generated and display similar postfragmentation density and exhaustive fragmentation. ©2011 American Physical Society

Molecular photoionization processes of astrophysical and aeronomical interest

NASA Technical Reports Server (NTRS)

Langhoff, P. W.

1985-01-01

An account is given of aspects of photoionization processes in molecules, with particular reference to recent theoretical and experimental studies of partial cross sections for production of specific final electronic states and of parent and fragment ions. Such cross sections help provide a basis for specifying the state of excitation of the ionized medium, are useful for estimating the kinetic energy distributions of photoejected electrons and fragment ions, provide parent-and fragment-ion yields, and clarify the possible origins of neutral fragments in highly excited rovibronic states. A descriptive account is given of photoionization phenomena, including tabulation of valence- and inner-shell potentials for some molecules of astrophysical and aeronomical interest. Cross sectional expressions are given. Various approximations currently employed in computational studies are described briefly, threshold laws and high-energy limits are indicated, and distinction is drawn between resonant and direct photoionization phenomena. Recent experimental and theoretical studies of partial photoionization cross sections in selected compounds of astrophysical and aeronomical relevance are described and discussed.

Dual small fragment plating improves screw-to-screw load sharing for mid-diaphyseal humeral fracture fixation: a finite element study.

PubMed

Kosmopoulos, Victor; Luedke, Colten; Nana, Arvind D

2015-01-01

A smaller humerus in some patients makes the use of a large fragment fixation plate difficult. Dual small fragment plate constructs have been suggested as an alternative. This study compares the biomechanical performance of three single and one dual plate construct for mid-diaphyseal humeral fracture fixation. Five humeral shaft finite element models (1 intact and 4 fixation) were loaded in torsion, compression, posterior-anterior (PA) bending, and lateral-medial (LM) bending. A comminuted fracture was simulated by a 1-cm gap. Fracture fixation was modelled by: (A) 4.5-mm 9-hole large fragment plate (wide), (B) 4.5-mm 9-hole large fragment plate (narrow), (C) 3.5-mm 9-hole small fragment plate, and (D) one 3.5-mm 9-hole small fragment plate and one 3.5-mm 7-hole small fragment plate. Model A showed the best outcomes in torsion and PA bending, whereas Model D outperformed the others in compression and LM bending. Stress concentrations were located near and around the unused screw holes for each of the single plate models and at the neck of the screws just below the plates for all the models studied. Other than in PA bending, Model D showed the best overall screw-to-screw load sharing characteristics. The results support using a dual small fragment locking plate construct as an alternative in cases where crutch weight-bearing (compression) tolerance may be important and where anatomy limits the size of the humerus bone segment available for large fragment plate fixation.

Distal radius fractures and the volar lunate facet fragment: Kirschner wire fixation in addition to volar-locked plating.

PubMed

Moore, Amy M; Dennison, David G

2014-06-01

The volar lunate facet fragment of a distal radius fracture may not be stabilized with volar-locked plating alone due to the small size and distal location of the fragment. Identification and stabilization of this small fragment is critical as unstable fixation may result in radiocarpal and radioulnar joint subluxation. The addition of spring wire fixation with volar plating can provide stable internal fixation of this critical fracture fragment. A retrospective review (2006-2011) identified nine patients with distal radius fractures with an associated volar lunate facet fragment that were treated with volar-locked plating and spring wire fixation of the volar lunate facet fragment. Radiographic indices, range of motion, grip strength, and postoperative Patient-related wrist evaluation (PRWE) scores were obtained to assess pain and function. All distal radius fractures healed, and the volar lunate facet fragment reduction was maintained. The mean follow-up was 54 weeks. Mean active range of motion was 46° wrist flexion, 51° wrist extension, 80° pronation, and 68° supination. The mean grip strength was 21 Kg, achieving 66 % of the uninjured limb. The average PRWE score was 17. No patient required removal of hardware or had evidence of tendon irritation. The addition of spring wire fixation to volar-locked plating provided stable fixation of the volar lunate facet fragment of distal radius fractures without complication. This technique addresses a limitation of volar-locked plating to control the small volar lunate facet fragment in distal radius fractures otherwise amenable to volar plating. A retrospective case series, Level IV.

Momentum transfer in the reactions of 13. 6-GeV/nucleon sup 16 O with copper

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cumming, J.B.; Chu, Y.Y.; Haustein, P.E.

1992-11-01

Mean ranges and forward-to-backward ratios have been determined by the thick-target, thick-catcher technique for fragments from the interaction of 13.6-GeV/nucleon {sup 16}O with copper. Experimental data are analyzed in terms of a two-step model and results are discussed in the context of limiting fragmentation and factorization and the predictions of a simple model for momentum transfer. Momenta imparted in the initial interactions of 13.6-GeV/nucleon {sup 16}O were inferred to be {similar to}12% lower than those for fragmentation of Cu by high-energy protons.

The Thick Level-Set model for dynamic fragmentation

DOE PAGES

Stershic, Andrew J.; Dolbow, John E.; Moës, Nicolas

2017-01-04

The Thick Level-Set (TLS) model is implemented to simulate brittle media undergoing dynamic fragmentation. This non-local model is discretized by the finite element method with damage represented as a continuous field over the domain. A level-set function defines the extent and severity of damage, and a length scale is introduced to limit the damage gradient. Numerical studies in one dimension demonstrate that the proposed method reproduces the rate-dependent energy dissipation and fragment length observations from analytical, numerical, and experimental approaches. In conclusion, additional studies emphasize the importance of appropriate bulk constitutive models and sufficient spatial resolution of the length scale.

Workflow Design Using Fragment Composition

NASA Astrophysics Data System (ADS)

Mosser, Sébastien; Blay-Fornarino, Mireille; France, Robert

The Service-Oriented Architecture (Soa) paradigm supports the assembly of atomic services to create applications that implement complex business processes. Assembly can be accomplished by service orchestrations defined by Soa architects. The Adore method allows Soa architects to model complex orchestrations of services by composing models of smaller orchestrations called orchestration fragments. The Adore method can also be used to weave fragments that address new concerns into existing application models. In this paper we illustrate how the Adore method can be used to separate and compose process aspects in a Soa design of the Car Crash Crisis Management System. The paper also includes a discussion of the benefits and limitations of the Adore method.

Astrometric and Photometric Analysis of the September 2008 ATV-1 Re-Entry Event

NASA Technical Reports Server (NTRS)

Mulrooney, Mark K.; Barker, Edwin S.; Maley, Paul D.; Beaulieu, Kevin R.; Stokely, Christopher L.

2008-01-01

NASA utilized Image Intensified Video Cameras for ATV data acquisition from a jet flying at 12.8 km. Afterwards the video was digitized and then analyzed with a modified commercial software package, Image Systems Trackeye. Astrometric results were limited by saturation, plate scale, and imposed linear plate solution based on field reference stars. Time-dependent fragment angular trajectories, velocities, accelerations, and luminosities were derived in each video segment. It was evident that individual fragments behave differently. Photometric accuracy was insufficient to confidently assess correlations between luminosity and fragment spatial behavior (velocity, deceleration). Use of high resolution digital video cameras in future should remedy this shortcoming.

Long-term demographic consequences of habitat fragmentation to a tropical understory bird community

USGS Publications Warehouse

Korfanta, N.M.; Newmark, W.D.; Kauffman, M.J.

2012-01-01

Tropical deforestation continues to cause population declines and local extinctions in centers of avian diversity and endemism. Although local species extinctions stem from reductions in demographic rates, little is known about how habitat fragmentation influences survival of tropical bird populations or the relative importance of survival and fecundity in ultimately shaping communities. We analyzed 22 years of mark-recapture data to assess how fragmentation influenced apparent survival, recruitment, and realized population growth rate within 22 forest understory bird species in the Usambara Mountains, Tanzania. This represents the first such effort, in either tropical or temperate systems, to characterize the effect of deforestation on avian survival across such a broad suite of species. Long-term demographic analysis of this suite of species experiencing the same fragmented environment revealed considerable variability in species' responses to fragmentation, in addition to general patterns that emerged from comparison among species. Across the understory bird community as a whole, we found significantly lower apparent survival and realized population growth rate in small fragments relative to large, demonstrating fragmentation effects to demographic rates long after habitat loss. Demographic rates were depressed across five feeding guilds, suggesting that fragmentation sensitivity was not limited to insectivores. Seniority analyses, together with a positive effect of fragmentation on recruitment, indicated that depressed apparent survival was the primary driver of population declines and observed extinctions. We also found a landscape effect, with lower vital rates in one mountain range relative to another, suggesting that fragmentation effects may add to other large-scale drivers of population decline. Overall, realized population growth rate (λ) estimates were < 1 for most species, suggesting that future population persistence even within large forest fragments is uncertain in this biodiversity hotspot.

Rational Design of High-Number dsDNA Fragments Based on Thermodynamics for the Construction of Full-Length Genes in a Single Reaction.

PubMed

Birla, Bhagyashree S; Chou, Hui-Hsien

2015-01-01

Gene synthesis is frequently used in modern molecular biology research either to create novel genes or to obtain natural genes when the synthesis approach is more flexible and reliable than cloning. DNA chemical synthesis has limits on both its length and yield, thus full-length genes have to be hierarchically constructed from synthesized DNA fragments. Gibson Assembly and its derivatives are the simplest methods to assemble multiple double-stranded DNA fragments. Currently, up to 12 dsDNA fragments can be assembled at once with Gibson Assembly according to its vendor. In practice, the number of dsDNA fragments that can be assembled in a single reaction are much lower. We have developed a rational design method for gene construction that allows high-number dsDNA fragments to be assembled into full-length genes in a single reaction. Using this new design method and a modified version of the Gibson Assembly protocol, we have assembled 3 different genes from up to 45 dsDNA fragments at once. Our design method uses the thermodynamic analysis software Picky that identifies all unique junctions in a gene where consecutive DNA fragments are specifically made to connect to each other. Our novel method is generally applicable to most gene sequences, and can improve both the efficiency and cost of gene assembly.

Direct access to dithiobenzoate RAFT agent fragmentation rate coefficients by ESR spin-trapping.

PubMed

Ranieri, Kayte; Delaittre, Guillaume; Barner-Kowollik, Christopher; Junkers, Thomas

2014-12-01

The β-scission rate coefficient of tert-butyl radicals fragmenting off the intermediate resulting from their addition to tert-butyl dithiobenzoate-a reversible addition-fragmentation chain transfer (RAFT) agent-is estimated via the recently introduced electron spin resonance (ESR)-trapping methodology as a function of temperature. The newly introduced ESR-trapping methodology is critically evaluated and found to be reliable. At 20 °C, a fragmentation rate coefficient of close to 0.042 s(-1) is observed, whereas the activation parameters for the fragmentation reaction-determined for the first time-read EA = 82 ± 13.3 kJ mol(-1) and A = (1.4 ± 0.25) × 10(13) s(-1) . The ESR spin-trapping methodology thus efficiently probes the stability of the RAFT adduct radical under conditions relevant for the pre-equilibrium of the RAFT process. It particularly indicates that stable RAFT adduct radicals are indeed formed in early stages of the RAFT poly-merization, at least when dithiobenzoates are employed as controlling agents as stipulated by the so-called slow fragmentation theory. By design of the methodology, the obtained fragmentation rate coefficients represent an upper limit. The ESR spin-trapping methodology is thus seen as a suitable tool for evaluating the fragmentation rate coefficients of a wide range of RAFT adduct radicals. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Comparative Population Genetic Structure of the Endangered Southern Brown Bandicoot, Isoodon obesulus, in Fragmented Landscapes of Southern Australia

PubMed Central

Li, You; Cooper, Steven J. B.; Lancaster, Melanie L.; Packer, Jasmin G.; Carthew, Susan M.

2016-01-01

Genetic connectivity is a key factor for maintaining the persistence of populations in fragmented landscapes. In highly modified landscapes such us peri-urban areas, organisms’ dispersal among fragmented habitat patches can be reduced due to the surrounding matrix, leading to subsequent decreased gene flow and increased potential extinction risk in isolated sub-populations. However, few studies have compared within species how dispersal/gene flow varies between regions and among different forms of matrix that might be encountered. In the current study, we investigated gene flow and dispersal in an endangered marsupial, the southern brown bandicoot (Isoodon obesulus) in a heavily modified peri-urban landscape in South Australia, Australia. We used 14 microsatellite markers to genotype 254 individuals which were sampled from 15 sites. Analyses revealed significant genetic structure. Our analyses also indicated that dispersal was mostly limited to neighbouring sites. Comparisons of these results with analyses of a different population of the same species revealed that gene flow/dispersal was more limited in this peri-urban landscape than in a pine plantation landscape approximately 400 km to the south-east. These findings increase our understanding of how the nature of fragmentation can lead to profound differences in levels of genetic connectivity among populations of the same species. PMID:27096952

Fragment-based screening by protein crystallography: successes and pitfalls.

PubMed

Chilingaryan, Zorik; Yin, Zhou; Oakley, Aaron J

2012-10-08

Fragment-based drug discovery (FBDD) concerns the screening of low-molecular weight compounds against macromolecular targets of clinical relevance. These compounds act as starting points for the development of drugs. FBDD has evolved and grown in popularity over the past 15 years. In this paper, the rationale and technology behind the use of X-ray crystallography in fragment based screening (FBS) will be described, including fragment library design and use of synchrotron radiation and robotics for high-throughput X-ray data collection. Some recent uses of crystallography in FBS will be described in detail, including interrogation of the drug targets β-secretase, phenylethanolamine N-methyltransferase, phosphodiesterase 4A and Hsp90. These examples provide illustrations of projects where crystallography is straightforward or difficult, and where other screening methods can help overcome the limitations of crystallography necessitated by diffraction quality.

Fragment-Based Screening by Protein Crystallography: Successes and Pitfalls

PubMed Central

Chilingaryan, Zorik; Yin, Zhou; Oakley, Aaron J.

2012-01-01

Fragment-based drug discovery (FBDD) concerns the screening of low-molecular weight compounds against macromolecular targets of clinical relevance. These compounds act as starting points for the development of drugs. FBDD has evolved and grown in popularity over the past 15 years. In this paper, the rationale and technology behind the use of X-ray crystallography in fragment based screening (FBS) will be described, including fragment library design and use of synchrotron radiation and robotics for high-throughput X-ray data collection. Some recent uses of crystallography in FBS will be described in detail, including interrogation of the drug targets β-secretase, phenylethanolamine N-methyltransferase, phosphodiesterase 4A and Hsp90. These examples provide illustrations of projects where crystallography is straightforward or difficult, and where other screening methods can help overcome the limitations of crystallography necessitated by diffraction quality. PMID:23202926

Fragment-based drug discovery and its application to challenging drug targets.

PubMed

Price, Amanda J; Howard, Steven; Cons, Benjamin D

2017-11-08

Fragment-based drug discovery (FBDD) is a technique for identifying low molecular weight chemical starting points for drug discovery. Since its inception 20 years ago, FBDD has grown in popularity to the point where it is now an established technique in industry and academia. The approach involves the biophysical screening of proteins against collections of low molecular weight compounds (fragments). Although fragments bind to proteins with relatively low affinity, they form efficient, high quality binding interactions with the protein architecture as they have to overcome a significant entropy barrier to bind. Of the biophysical methods available for fragment screening, X-ray protein crystallography is one of the most sensitive and least prone to false positives. It also provides detailed structural information of the protein-fragment complex at the atomic level. Fragment-based screening using X-ray crystallography is therefore an efficient method for identifying binding hotspots on proteins, which can then be exploited by chemists and biologists for the discovery of new drugs. The use of FBDD is illustrated here with a recently published case study of a drug discovery programme targeting the challenging protein-protein interaction Kelch-like ECH-associated protein 1:nuclear factor erythroid 2-related factor 2. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Geographic variation in cowbird distribution, abundance, and parasitism

USGS Publications Warehouse

Morrison, M.L.; Hahn, D.C.; George, T. Luke; Dobkin, David S.

2002-01-01

We evaluated geographical patterns in the abundance and distribution of Brown-headed Cowbirds (Molothrus ater), and in the frequency of cowbird parasitism, across North America in relation to habitat fragmentation. We found no distinctive parasitism patterns at the national or even regional scales, but the species is most abundant in the Great Plains, the heart of their original range, and least common in the southeastern U.S. This situation is dynamic, because both the Brown-headed and two other cowbird species are actively expanding their ranges in the southern U.S. We focused almost entirely in this paper on the Brown-headed Cowbird, because it is the only endemic North American cowbird, its distribution is much wider, and it has been much more intensively studied. We determined that landscape is the most meaningful unit of scale for comparing cowbird parasitism patterns as, for example, in comparisons of northeastern and central hardwood forests within agricultural matrices, and suburbanized areas versus western coniferous forests. We concluded that cowbird parasitism patterns were broadly similar within all landscapes. Even comparisons between prominently dissimilar landscapes, such as hardwoods in agriculture and suburbia versus coniferous forest, display a striking similarity in the responses of cowbirds. Our review clearly indicated that proximity of feeding areas is the key factor influencing presence and parasitism patterns within the landscape. We considered intensity of landscape fragmentation from forest-dominated landscapes altered in a forest management context to fragmentation characterized by mixed suburbanization or agricultural development. Our review consistently identified an inverse relationship between extent of forest cover across the landscape and cowbird presence. Invariably, the variation seen in parasitism frequencies within a region was at least partially explained as a response to changes in forest cover. The most salient geographic aspect of cowbirds' response to landscape fragmentation is the time since fragmentation occurred. Eastern landscapes generally experienced 200 years ago the development and fragmentation that western landscapes experienced less than 75 years ago. Consequently, there is a broad east-west contrast in which more numerous human settlements and smaller unbroken forest stands are found in the East, a difference that permits cowbirds to be more pervasive and ubiquitous. The locality of suitable feeding areas is a hallmark trait of the cowbirds' strategy in exploiting specific forest fragments. Host abundance influences parasitism patterns only secondarily at the landscape scale. These two limiting factors come into play differently in different landscapes. For example, cowbird abundance in unbroken forested landscapes are limited primarily by the availability of foraging areas rather than by host density, whereas cowbirds are limited primarily by host availability in landscapes that are extensively fragmented with feeding areas.

Budgeting in Tertiary Educational Institutions: An Analysis.

ERIC Educational Resources Information Center

Shand, D. A.

1982-01-01

Administrators should examine the role and limitations of their budgeting systems and consider overemphasis on control of spending rather than efficiency or effectiveness, limitations on cash-based budgeting, need for information on operating costs, and the short range and fragmented nature of most budgets. (MSE)

Atmospheric-pressure ionization and fragmentation of peptides by solution-cathode glow discharge

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schwartz, Andrew J.; Shelley, Jacob T.; Walton, Courtney L.

Modern “-omics” (e.g., proteomics, glycomics, metabolomics, etc.) analyses rely heavily on electrospray ionization and tandem mass spectrometry to determine the structural identity of target species. Unfortunately, these methods are limited to specialized mass spectrometry instrumentation. Here in this paper, a novel approach is described that enables ionization and controlled, tunable fragmentation of peptides at atmospheric pressure. In the new source, a direct-current plasma is sustained between a tapered metal rod and a flowing sample-containing solution. As the liquid stream contacts the electrical discharge, peptides from the solution are volatilized, ionized, and fragmented. At high discharge currents (e.g., 70 mA), electrospray-likemore » spectra are observed, dominated by singly and doubly protonated molecular ions. At lower currents (35 mA), many peptides exhibit extensive fragmentation, with a-, b-, c-, x-, and y-type ion series present as well as complex fragments, such as d-type ions, not previously observed with atmospheric-pressure dissociation. Though the mechanism of fragmentation is currently unclear, observations indicate it could result from the interaction of peptides with gas-phase radicals or ultraviolet radiation generated within the plasma.« less

Atmospheric-pressure ionization and fragmentation of peptides by solution-cathode glow discharge

DOE PAGES

Schwartz, Andrew J.; Shelley, Jacob T.; Walton, Courtney L.; ...

2016-06-27

Modern “-omics” (e.g., proteomics, glycomics, metabolomics, etc.) analyses rely heavily on electrospray ionization and tandem mass spectrometry to determine the structural identity of target species. Unfortunately, these methods are limited to specialized mass spectrometry instrumentation. Here in this paper, a novel approach is described that enables ionization and controlled, tunable fragmentation of peptides at atmospheric pressure. In the new source, a direct-current plasma is sustained between a tapered metal rod and a flowing sample-containing solution. As the liquid stream contacts the electrical discharge, peptides from the solution are volatilized, ionized, and fragmented. At high discharge currents (e.g., 70 mA), electrospray-likemore » spectra are observed, dominated by singly and doubly protonated molecular ions. At lower currents (35 mA), many peptides exhibit extensive fragmentation, with a-, b-, c-, x-, and y-type ion series present as well as complex fragments, such as d-type ions, not previously observed with atmospheric-pressure dissociation. Though the mechanism of fragmentation is currently unclear, observations indicate it could result from the interaction of peptides with gas-phase radicals or ultraviolet radiation generated within the plasma.« less

Searching for substructures in fragment spaces.

PubMed

Ehrlich, Hans-Christian; Volkamer, Andrea; Rarey, Matthias

2012-12-21

A common task in drug development is the selection of compounds fulfilling specific structural features from a large data pool. While several methods that iteratively search through such data sets exist, their application is limited compared to the infinite character of molecular space. The introduction of the concept of fragment spaces (FSs), which are composed of molecular fragments and their connection rules, made the representation of large combinatorial data sets feasible. At the same time, search algorithms face the problem of structural features spanning over multiple fragments. Due to the combinatorial nature of FSs, an enumeration of all products is impossible. In order to overcome these time and storage issues, we present a method that is able to find substructures in FSs without explicit product enumeration. This is accomplished by splitting substructures into subsubstructures and mapping them onto fragments with respect to fragment connectivity rules. The method has been evaluated on three different drug discovery scenarios considering the exploration of a molecule class, the elaboration of decoration patterns for a molecular core, and the exhaustive query for peptides in FSs. FSs can be searched in seconds, and found products contain novel compounds not present in the PubChem database which may serve as hints for new lead structures.

Separation efficiency of free-solution conjugated electrophoresis with drag-tags incorporating a synthetic amino acid.

PubMed

Seo, Kyung-Ho; Chu, Hun-Su; Yoo, Tae Hyeon; Lee, Sun-Gu; Won, Jong-In

2016-03-01

DNA sequencing or separation by conventional capillary electrophoresis with a polymer matrix has some inherent drawbacks, such as the expense of polymer matrix and limitations in sequencing read length. As DNA fragments have a linear charge-to-friction ratio in free solution, DNA fragments cannot be separated by size. However, size-based separation of DNA is possible in free-solution conjugate electrophoresis (FSCE) if a "drag-tag" is attached to DNA fragments because the tag breaks the linear charge-to-friction scaling. Although several previous studies have demonstrated the feasibility of DNA separation by free-solution conjugated electrophoresis, generation of a monodisperse drag-tag and identification of a strong, site-specific conjugation method between a DNA fragment and a drag-tag are challenges that still remain. In this study, we demonstrate an efficient FSCE method by conjugating a biologically synthesized elastin-like polypeptide (ELP) and green fluorescent protein (GFP) to DNA fragments. In addition, to produce strong and site-specific conjugation, a methionine residue in drag-tags is replaced with homopropargylglycine (Hpg), which can be conjugated specifically to a DNA fragment with an azide site. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The Split Nucleus of Comet Wilson (C/1986 P1 = 1987 VII).

NASA Astrophysics Data System (ADS)

Meech, Karen J.; Knopp, Graham P.; Farnham, Tony L.; Green, Daniel

1995-07-01

We present CCD observations of Comet Wilson (C/1986 P1 = 1987 VII) from 26 nights during the time period 1986 October to 1991 February, which brackets perihelion, During the observing run of 1988 February, the comet was observed to have split into two fragments. Our broadband CCD photometry, along with photometry from the International Cometary Quarterly, shows a steady decline in brightness of Comet Wilson post-perihelion, with an outburst between heliocentric distances r = 2.8 and 3.3 AU during 1987 October and November. By r ≈ 7 AU, the fragment had faded with respect to the parent and was no longer centrally condensed. A brightness limit of mR ≈ 25, when the comet was at r = 12.65 AU, constrains the primary nucleus to have a maximum radius between 5 and 7 km, assuming an albedo of 0.04. The accuracy of direct orbital solutions for the parent body and fragment to determine the time of splitting was limited by the presence of significant nongravitational forces and the limited fragment orbital coverage. We used the relative position of the fragment with respect to the parent to calculate a time of splitting which was consistent with the time of the observed outburst. We discuss the possible causes of the splitting. The coma of Comet Wilson was observed to have a surface-brightness profile which fell off as p-1 (characteristic of a canonical steady-state coma under the influence of radiation pressure) for all of the data with the exception of the data taken during 1987 November when the gradient was p-1.3 . This steeper slope was probably caused by the injection of new material into the coma during the outburst. During 1986 October, there was a break in the surface-brightness profile slope which may be interpreted as the distance at which grains are swept into the tail. The profiles suggested grain velocities of a few x 10 2 to 10 m sec -1 for grains between 1 and a few hundred micrometers. Finson-Probstein dust modeling showed that ejection of grains began preperihelion at r gt; 6 AU, peaking near perihelion with a very rapid decline thereafter, with an outburst occurring roughly 170 days post-perihelion. We found no evidence for the existence of discrete features in the coma. Both the primary and fragment nuclei grew redder with time, with the fragment being redder than the parent, consistent with the picture of smaller grains leaving the field of view of the aperture more quickly than larger grains.

High-throughput analysis of the protein sequence-stability landscape using a quantitative "yeast surface two-hybrid" system and fragment reconstitution

PubMed Central

Dutta, Sanjib; Koide, Akiko; Koide, Shohei

2008-01-01

Stability evaluation of many mutants can lead to a better understanding of the sequence determinants of a structural motif and of factors governing protein stability and protein evolution. The traditional biophysical analysis of protein stability is low throughput, limiting our ability to widely explore the sequence space in a quantitative manner. In this study, we have developed a high-throughput library screening method for quantifying stability changes, which is based on protein fragment reconstitution and yeast surface display. Our method exploits the thermodynamic linkage between protein stability and fragment reconstitution and the ability of the yeast surface display technique to quantitatively evaluate protein-protein interactions. The method was applied to a fibronectin type III (FN3) domain. Characterization of fragment reconstitution was facilitated by the co-expression of two FN3 fragments, thus establishing a "yeast surface two-hybrid" method. Importantly, our method does not rely on competition between clones and thus eliminates a common limitation of high-throughput selection methods in which the most stable variants are predominantly recovered. Thus, it allows for the isolation of sequences that exhibits a desired level of stability. We identified over one hundred unique sequences for a β-bulge motif, which was significantly more informative than natural sequences of the FN3 family in revealing the sequence determinants for the β-bulge. Our method provides a powerful means to rapidly assess stability of many variants, to systematically assess contribution of different factors to protein stability and to enhance protein stability. PMID:18674545

A probabilistic and continuous model of protein conformational space for template-free modeling.

PubMed

Zhao, Feng; Peng, Jian; Debartolo, Joe; Freed, Karl F; Sosnick, Tobin R; Xu, Jinbo

2010-06-01

One of the major challenges with protein template-free modeling is an efficient sampling algorithm that can explore a huge conformation space quickly. The popular fragment assembly method constructs a conformation by stringing together short fragments extracted from the Protein Data Base (PDB). The discrete nature of this method may limit generated conformations to a subspace in which the native fold does not belong. Another worry is that a protein with really new fold may contain some fragments not in the PDB. This article presents a probabilistic model of protein conformational space to overcome the above two limitations. This probabilistic model employs directional statistics to model the distribution of backbone angles and 2(nd)-order Conditional Random Fields (CRFs) to describe sequence-angle relationship. Using this probabilistic model, we can sample protein conformations in a continuous space, as opposed to the widely used fragment assembly and lattice model methods that work in a discrete space. We show that when coupled with a simple energy function, this probabilistic method compares favorably with the fragment assembly method in the blind CASP8 evaluation, especially on alpha or small beta proteins. To our knowledge, this is the first probabilistic method that can search conformations in a continuous space and achieves favorable performance. Our method also generated three-dimensional (3D) models better than template-based methods for a couple of CASP8 hard targets. The method described in this article can also be applied to protein loop modeling, model refinement, and even RNA tertiary structure prediction.

Production of exotic nuclei in projectile fragmentation at relativistic and Fermi energies

NASA Astrophysics Data System (ADS)

Ogul, R.; Ergun, A.; Buyukcizmeci, N.

2017-02-01

Isotopic distributions of projectile fragmentation in peripheral heavy ion collisions of 86Kr on 112Sn are calculated within the statistical multifragmentation model. Obtained data are compared to the experimental cross section measurements. We show the enhancement in the production of neutron-rich isotopes close to the projectile, observed in the experiments. Our results show the universality of the limitation of the excitation energy induced in the projectile residues.

String Fragmentation Model in Space Radiation Problems

NASA Technical Reports Server (NTRS)

Tang, Alfred; Johnson, Eloise (Editor); Norbury, John W.; Tripathi, R. K.

2002-01-01

String fragmentation models such as the Lund Model fit experimental particle production cross sections very well in the high-energy limit. This paper gives an introduction of the massless relativistic string in the Lund Model and shows how it can be modified with a simple assumption to produce formulas for meson production cross sections for space radiation research. The results of the string model are compared with inclusive pion production data from proton-proton collision experiments.

Characterization of drop aerodynamic fragmentation in the bag and sheet-thinning regimes by crossed-beam, two-view, digital in-line holography

DOE PAGES

Guildenbecher, Daniel R.; Gao, Jian; Chen, Jun; ...

2017-04-19

When a spherical liquid drop is subjected to a step change in relative gas velocity, aerodynamic forces lead to drop deformation and possible breakup into a number of secondary fragments. In order to investigate this flow, a digital in-line holography (DIH) diagnostic is proposed which enables rapid quantification of spatial statistics with limited experimental repetition. To overcome the high uncertainty in the depth direction experienced in previous applications of DIH, a crossed-beam, two-view configuration is introduced. With appropriate calibration, this diagnostic is shown to provide accurate quantification of fragment sizes, three-dimensional positions and three-component velocities in a large measurement volume.more » We apply these capabilities in order to investigate the aerodynamic breakup of drops at two non-dimensional Weber numbers, We, corresponding to the bag (We = 14) and sheet-thinning (We = 55) regimes. Ensemble average results show the evolution of fragment size and velocity statistics during the course of breakup. Our results indicate that mean fragment sizes increase throughout the course of breakup. For the bag breakup case, the evolution of a multi-mode fragment size probability density is observed. This is attributed to separate fragmentation mechanisms for the bag and rim structures. In contrast, for the sheet-thinning case, the fragment size probability density shows only one distinct peak indicating a single fragmentation mechanism. Compared to previous related investigations of this flow, many orders of magnitude more fragments are measured per condition, resulting in a significant improvement in data fidelity. For this reason, this experimental dataset is likely to provide new opportunities for detailed validation of analytic and computational models of this flow.« less

Strong and nonlinear effects of fragmentation on ecosystem service provision at multiple scales

NASA Astrophysics Data System (ADS)

Mitchell, Matthew G. E.; Bennett, Elena M.; Gonzalez, Andrew

2015-09-01

Human actions, such as converting natural land cover to agricultural or urban land, result in the loss and fragmentation of natural habitat, with important consequences for the provision of ecosystem services. Such habitat loss is especially important for services that are supplied by fragments of natural land cover and that depend on flows of organisms, matter, or people across the landscape to produce benefits, such as pollination, pest regulation, recreation and cultural services. However, our quantitative knowledge about precisely how different patterns of landscape fragmentation might affect the provision of these types of services is limited. We used a simple, spatially explicit model to evaluate the potential impact of natural land cover loss and fragmentation on the provision of hypothetical ecosystem services. Based on current literature, we assumed that fragments of natural land cover provide ecosystem services to the area surrounding them in a distance-dependent manner such that ecosystem service flow depended on proximity to fragments. We modeled seven different patterns of natural land cover loss across landscapes that varied in the overall level of landscape fragmentation. Our model predicts that natural land cover loss will have strong and unimodal effects on ecosystem service provision, with clear thresholds indicating rapid loss of service provision beyond critical levels of natural land cover loss. It also predicts the presence of a tradeoff between maximizing ecosystem service provision and conserving natural land cover, and a mismatch between ecosystem service provision at landscape versus finer spatial scales. Importantly, the pattern of landscape fragmentation mitigated or intensified these tradeoffs and mismatches. Our model suggests that managing patterns of natural land cover loss and fragmentation could help influence the provision of multiple ecosystem services and manage tradeoffs and synergies between services across different human-dominated landscapes.

The fragmentation instability of a black hole with f( R) global monopole under GUP

NASA Astrophysics Data System (ADS)

Chen, Lingshen; Cheng, Hongbo

2018-03-01

Having studied the fragmentation of the black holes containing f( R) global monopole under the generalized uncertainty principle (GUP), we show the influences from this kind of monopole, f( R) theory, and GUP on the evolution of black holes. We focus on the possibility that the black hole breaks into two parts by means of the second law of thermodynamics. We derive the entropies of the initial black hole and the broken parts while the generalization of Heisenberg's uncertainty principle is introduced. We find that the f( R) global monopole black hole keeps stable instead of splitting without the generalization because the entropy difference is negative. The fragmentation of the black hole will happen if the black hole entropies are limited by the GUP and the considerable deviation from the general relativity leads to the case that the mass of one fragmented black hole is smaller and the other one's mass is larger.

Corono-radicular biological restoration of maxillary central incisors by direct method.

PubMed

Aggarwal, Sonia; Sahoo, Sujit Ranjan; Pandharkar, Kartik

2014-11-01

This case report refers to the esthetic and functional restorations of extensively damaged maxillary central incisors with dental caries in a 32-year-old woman, with the use of posts and crowns made from natural extracted teeth. Proper restoration of such teeth with the use of natural teeth fragments are known as "biological restoration." Biological restorations can be done by using the fragments of the patients own tooth and if that is not available, tooth fragment can be obtained from an extracted tooth. These biological posts and crowns present a low cost option and an alternative technique for the morphofunctional recovery of extensively damaged teeth. There are limitations with the use of natural extracted teeth (homogenous bonding) for restoration such as the difficulty of finding teeth with a similar color and shape as that of the destroyed element, or patient may refuse to accept a tooth fragment from another patient, which prevents execution of the restoration.

Time-resolved ion imaging at free-electron lasers using TimepixCam.

PubMed

Fisher-Levine, Merlin; Boll, Rebecca; Ziaee, Farzaneh; Bomme, Cédric; Erk, Benjamin; Rompotis, Dimitrios; Marchenko, Tatiana; Nomerotski, Andrei; Rolles, Daniel

2018-03-01

The application of a novel fast optical-imaging camera, TimepixCam, to molecular photoionization experiments using the velocity-map imaging technique at a free-electron laser is described. TimepixCam is a 256 × 256 pixel CMOS camera that is able to detect and time-stamp ion hits with 20 ns timing resolution, thus making it possible to record ion momentum images for all fragment ions simultaneously and avoiding the need to gate the detector on a single fragment. This allows the recording of significantly more data within a given amount of beam time and is particularly useful for pump-probe experiments, where drifts, for example, in the timing and pulse energy of the free-electron laser, severely limit the comparability of pump-probe scans for different fragments taken consecutively. In principle, this also allows ion-ion covariance or coincidence techniques to be applied to determine angular correlations between fragments.

Urban habitat fragmentation and genetic population structure of bobcats in coastal southern California

USGS Publications Warehouse

Ruell, E.W.; Riley, S.P.D.; Douglas, M.R.; Antolin, M.F.; Pollinger, J.R.; Tracey, J.A.; Lyren, L.M.; Boydston, E.E.; Fisher, R.N.; Crooks, K.R.

2012-01-01

Although habitat fragmentation is recognized as a primary threat to biodiversity, the effects of urban development on genetic population structure vary among species and landscapes and are not yet well understood. Here we use non-invasive genetic sampling to compare the effects of fragmentation by major roads and urban development on levels of dispersal, genetic diversity, and relatedness between paired bobcat populations in replicate landscapes in coastal southern California. We hypothesized that bobcat populations in sites surrounded by urbanization would experience reduced functional connectivity relative to less isolated nearby populations. Our results show that bobcat genetic population structure is affected by roads and development but not always as predicted by the degree that these landscape features surround fragments. Instead, we suggest that urban development may affect functional connectivity between bobcat populations more by limiting the number and genetic diversity of source populations of migrants than by creating impermeable barriers to dispersal.

Tandem mass spectrometry of large biomolecule ions by blackbody infrared radiative dissociation.

PubMed

Price, W D; Schnier, P D; Williams, E R

1996-03-01

A new method for the dissociation of large ions formed by electrospray ionization is demonstrated. Ions trapped in a Fourier transform mass spectrometer at pressures below 10(-)(8) Torr are dissociated by elevating the vacuum chamber to temperatures up to 215 °C. Rate constants for dissociation are measured and found to be independent of pressure below 10(-)(7) Torr. This indicates that the ions are activated by absorption of blackbody radiation emitted from the chamber walls. Dissociation efficiencies as high as 100% are obtained. There is no apparent mass limit to this method; ions as large as ubiquitin (8.6 kDa) are readily dissociated. Thermally stable ions, such as melittin 3+ (2.8 kDa), did not dissociate at temperatures up to 200 °C. This method is highly selective for low-energy fragmentation, from which limited sequence information can be obtained. From the temperature dependence of the dissociation rate constants, Arrhenius activation energies in the low-pressure limit are obtained. The lowest energy dissociation processes for the singly and doubly protonated ions of bradykinin are loss of NH(3) and formation of the b(2)/y(7) complementary pair, with activation energies of 1.3 and 0.8 eV, respectively. No loss of NH(3) is observed for the doubly protonated ion; some loss of H(2)O occurs. These results show that charge-charge interactions not only lower the activation energy for dissociation but also can dramatically change the fragmentation, most likely through changes in the gas-phase conformation of the ion. Dissociation of ubiquitin ions produces fragmentation similar to that obtained by IRMPD and SORI-CAD. Higher charge state ions dissociate to produce y and b ions; the primary fragmentation process for low charge state ions is loss of H(2)O.

Coulomb fission in dielectric dication clusters: experiment and theory on steps that may underpin the electrospray mechanism.

PubMed

Chen, Xiaojing; Bichoutskaia, Elena; Stace, Anthony J

2013-05-16

A series of five molecular dication clusters, (H2O)n(2+), (NH3)n(2+), (CH3CN)n(2+), (C5H5N)n(2+), and (C6H6)n(2+), have been studied for the purpose of identifying patterns of behavior close to the Rayleigh instability limit where the clusters might be expected to exhibit Coulomb fission. Experiments show that the instability limit for each dication covers a range of sizes and that on a time scale of 10(-4) s ions close to the limit can undergo either Coulomb fission or neutral evaporation. The observed fission pathways exhibit considerable asymmetry in the sizes of the charged fragments, and are associated with kinetic (ejection) energies of ~0.9 eV. Coulomb fission has been modeled using a theory recently formulated to describe how charged particles of dielectric materials interact with one another (Bichoutskaia et al. J. Chem. Phys. 2010, 133, 024105). The calculated electrostatic interaction energy between separating fragments accounts for the observed asymmetric fragmentation and for the magnitudes of the measured ejection energies. The close match between theory and experiment suggests that a significant fraction of excess charge resides on the surfaces of the fragment ions. The experiments provided support for a fundamental step in the electrospray ionization (ESI) mechanism, namely the ejection from droplets of small solvated charge carriers. At the same time, the theory shows how water and acetonitrile may behave slightly differently as ESI solvents. However, the theory also reveals deficiencies in the point-charge image-charge model that has previously been used to quantify Coulomb fission in the electrospray process.

In-gel multiple displacement amplification of long DNA fragments diluted to the single molecule level.

PubMed

Michikawa, Yuichi; Sugahara, Keisuke; Suga, Tomo; Ohtsuka, Yoshimi; Ishikawa, Kenichi; Ishikawa, Atsuko; Shiomi, Naoko; Shiomi, Tadahiro; Iwakawa, Mayumi; Imai, Takashi

2008-12-15

The isolation and multiple genotyping of long individual DNA fragments are needed to obtain haplotype information for diploid organisms. Limiting dilution of sample DNA followed by multiple displacement amplification is a useful technique but is restricted to short (<5 kb) DNA fragments. In the current study, a novel modification was applied to overcome these problems. A limited amount of cellular DNA was carefully released from intact cells into a mildly heated alkaline agarose solution and mixed thoroughly. The solution was then gently aliquoted and allowed to solidify while maintaining the integrity of the diluted DNA. Exogenously provided Phi29 DNA polymerase was used to perform consistent genomic amplification with random hexameric oligonucleotides within the agarose gels. Simple heat melting of the gel allowed recovery of the amplified materials in a solution of the polymerase chain reaction (PCR)-ready form. The haplotypes of seven SNPs spanning 240 kb of the DNA surrounding the human ATM gene region on chromosome 11 were determined for 10 individuals, demonstrating the feasibility of this new method.

Robustness of optimal random searches in fragmented environments

NASA Astrophysics Data System (ADS)

Wosniack, M. E.; Santos, M. C.; Raposo, E. P.; Viswanathan, G. M.; da Luz, M. G. E.

2015-05-01

The random search problem is a challenging and interdisciplinary topic of research in statistical physics. Realistic searches usually take place in nonuniform heterogeneous distributions of targets, e.g., patchy environments and fragmented habitats in ecological systems. Here we present a comprehensive numerical study of search efficiency in arbitrarily fragmented landscapes with unlimited visits to targets that can only be found within patches. We assume a random walker selecting uniformly distributed turning angles and step lengths from an inverse power-law tailed distribution with exponent μ . Our main finding is that for a large class of fragmented environments the optimal strategy corresponds approximately to the same value μopt≈2 . Moreover, this exponent is indistinguishable from the well-known exact optimal value μopt=2 for the low-density limit of homogeneously distributed revisitable targets. Surprisingly, the best search strategies do not depend (or depend only weakly) on the specific details of the fragmentation. Finally, we discuss the mechanisms behind this observed robustness and comment on the relevance of our results to both the random search theory in general, as well as specifically to the foraging problem in the biological context.

Effects of natural forest fragmentation on a Hawaiian spider community

USGS Publications Warehouse

Vandergast, Amy; Gillespie, Rosemary G.

2004-01-01

The kipuka system, a network of forest fragments surrounded by lava flows on the island of Hawaii, offers an opportunity to study the natural, long-term fragmentation of a native ecosystem. We examined the impacts of habitat edges upon the community structure of nocturnally active native spiders, primarily in the genus Tetragnatha. We measured plant and spider species distributions across the edges of four small fragments and one large continuously forested area that were surrounded by a lava flow in 1855. Results indicated that an ???20 m edge ecotone surrounds core forest habitat. Spider community structure changed across the edge, with a decrease in total species richness and diversity at the forest/lava boundary, and a change in the dominant taxon from native Tetragnatha (Tetragnathidae) to native Cyclosa (Araneidae). Severe habitat restrictions were found for some spider species. In addition, nearly all of the spiders captured were endemic species, and the few introduced species were limited to the younger and more open lava flows. Our results suggest that species responses to edges can vary, and that core habitat specialists may decline in fragmented conditions.

Ultrasensitive direct impedimetric immunosensor for detection of serum HER2.

PubMed

Sharma, Shikha; Zapatero-Rodríguez, Julia; Saxena, Rahul; O'Kennedy, Richard; Srivastava, Sudha

2018-05-30

Assesment of human epidermal growth factor receptor 2 status is a key factor prompting definitive treatment decisions that help in reducing mortality rates associated with breast cancer. In this article, highly sensitive and low-cost impedimetric immunosensor using single-chain fragment variable antibody fragments was developed for quantitative detection of human epidermal growth factor receptor 2 from serum employing gold nanoparticle-modified disposable screen-printed carbon electrodes. The gold nanoparticles facilitate fast electron transfer and offer a biocompatible surface for immobilization of small antibody fragments in an oriented manner, resulting in improved antigen binding efficiency. The single-chain fragment variable antibody fragment-modified screen printed immunosensor exhibits wide dynamic range of 0.01-100 ng mL -1 and detection limit of 0.01 ng mL -1 . The advantages offered by this platform in terms of high sensitivity, broad dynamic range and low-cost demonstrates great potential for improved monitoring of human epidermal growth factor receptor 2 levels for the management of breast and other cancers. Copyright © 2018 Elsevier B.V. All rights reserved.

Collision-induced fragmentation of negative ions from N-linked glycans derivatized with 2-aminobenzoic acid.

PubMed

Harvey, David J

2005-05-01

N-Linked glycans from bovine ribonuclease B, chicken ovalbumin, bovine fetuin, porcine thyroglobulin and human alpha(1)-acid glycoprotein were derivatized with 2-aminobenzoic acid by reductive amination and their tandem mass spectra were recorded by negative ion electrospray ionization with a quadrupole time-of-flight mass spectrometer. Derivatives were also prepared from 2-amino-5-methyl- and 2-amino-4,5-dimethoxybenzoic acid in order to confirm the identity of fragment ions containing the reducing terminus. Major fragments from the [M - H](-) ions from the neutral glycans retained the derivative (Y-type cleavages) and provided information on sequence and branching. Other major fragments were products of A-type cross-ring cleavages giving information on antenna structure. Singly doubly and triply charged ions were formed from sialylated glycans. They produced major fragments by loss of sialic acid and a series of singly charged ions that were similar to those from the neutral analogues. Doubly charge ions were also produced by the neutral glycans and were fragmented to form product ions with one and two charges. Again, the fragment ions with a single charge were similar to those from the singly charged parents, but branching information was less obvious because of the occurrence of more abundant ions produced by multiple cleavages. Detection limits were around 200 fmol (3 : 1 signal-to-noise ratio). Copyright 2005 John Wiley & Sons, Ltd.

Age differences in implicit memory: conceptual, perceptual, or methodological?

PubMed

Mitchell, David B; Bruss, Peter J

2003-12-01

The authors examined age differences in conceptual and perceptual implicit memory via word-fragment completion, word-stem completion, category exemplar generation, picture-fragment identification, and picture naming. Young, middle-aged, and older participants (N = 60) named pictures and words at study. Limited test exposure minimized explicit memory contamination, yielding no reliable age differences and equivalent cross-format effects. In contrast, explicit memory and neuropsychological measures produced significant age differences. In a follow-up experiment, 24 young adults were informed a priori about implicit testing. Their priming was equivalent to the main experiment, showing that test trial time restrictions limit explicit memory strategies. The authors concluded that most implicit memory processes remain stable across adulthood and suggest that explicit contamination be rigorously monitored in aging studies.

Analytic Ballistic Performance Model of Whipple Shields

NASA Technical Reports Server (NTRS)

Miller, J. E.; Bjorkman, M. D.; Christiansen, E. L.; Ryan, S. J.

2014-01-01

The dual-wall Whipple shield is the shield of choice for lightweight, long-duration flight. The shield uses an initial sacrificial wall to initiate fragmentation and melt an impacting threat that expands over a void before hitting a subsequent shield wall of a critical component. The key parameters to this type of shield are the rear wall and its mass which stops the debris, as well as the minimum pressure generated under threat particle impact of the sacrificial wall and the amount of void that is available for expansion. Ensuring the minimum pressure is sufficiently high to achieve large scale fragmentation/melt of the threat particle enables the expansion of the threat and reduces the momentum flux of the debris on the rear wall. Three key factors in the minimum pressure achieved are the thickness of the sacrificial wall relative to the characteristic dimension of the impacting particle, the density and material cohesion contrast of the sacrificial wall relative to the threat particle and the impact speed. The mass of the rear wall and the sacrificial wall are desirable to minimize for launch costs and dynamic concerns making it important to have an understanding of the effects of density contrast and impact speed. In this paper a fourth key parameter is identified related to fragmentation, which corresponds to the ratio of the size of the projectile relative to the transition from brittle to ductile hole growth in the projectile. Ballistic limit equations have been developed to define the failure limits of a MMOD shield, generally in terms of projectile diameter (or mass), impact velocity, and angle. Within the range of impact velocities relevant for Earth-orbiting spacecraft, three distinct regions of penetration phenomenology have been identified for Whipple shields: center dot Low velocity: the projectile is eroded (and possibly deformed) during its passage through the bumper plate, but is not fragmented. Thus, perforation of the rear wall is by a fragment with a mass and speed equal to or less than the original impactor. center dot Intermediate (shatter) velocity: impact velocities are sufficient to induce projectile fragmentation upon impact with the bumper plate, resulting in a coarse debris cloud with large solid fragments. Increasing velocity within the shatter regime results in increased fragmentation, and eventually melting, of the projectile and bumper fragments, generating a finer and more evenly dispersed debris cloud. Failure of the rear wall is a complicated combination of modes observed at low- and hypervelocity. center dot Hypervelocity: the projectile and holed-out bumper material is completely, or nearly completely, melted and/or vaporized by the initial impact. The resultant debris cloud impacts over a dispersed area of the rear wall, loading it impulsively and inducing failure through rupture or petalling. While each of these regimes are well observed with extensive empirical methods to describe these regions, differences in impactor materials, configurations of shields and questions about the limitations of the attainable impact speeds have left questions that are difficult to answer from completely empirical methods.

Genetic connectivity of the moth pollinated tree Glionnetia sericea in a highly fragmented habitat.

PubMed

Finger, Aline; Kaiser-Bunbury, Christopher N; Kettle, Chris J; Valentin, Terence; Ghazoul, Jaboury

2014-01-01

Long-distance gene flow is thought to be one prerequisite for the persistence of plant species in fragmented environments. Human influences have led to severe fragmentation of native habitats in the Seychelles islands, with many species surviving only in small and isolated populations. The endangered Seychelles endemic tree Glionnetia sericea is restricted to altitudes between 450 m and 900 m where the native forest vegetation has been largely lost and replaced with exotic invasives over the last 200 years. This study explores the genetic and ecological consequences of population fragmentation in this species by analysing patterns of genetic diversity in a sample of adults, juveniles and seeds, and by using controlled pollination experiments. Our results show no decrease in genetic diversity and no increase in genetic structuring from adult to juvenile cohorts. Despite significant inbreeding in some populations, there is no evidence of higher inbreeding in juvenile cohorts relative to adults. A Bayesian structure analysis and a tentative paternity analysis indicate extensive historical and contemporary gene flow among remnant populations. Pollination experiments and a paternity analysis show that Glionnetia sericea is self-compatible. Nevertheless, outcrossing is present with 7% of mating events resulting from pollen transfer between populations. Artificial pollination provided no evidence for pollen limitation in isolated populations. The highly mobile and specialized hawkmoth pollinators (Agrius convolvuli and Cenophodes tamsi; Sphingidae) appear to promote extensive gene flow, thus mitigating the potential negative ecological and genetic effects of habitat fragmentation in this species. We conclude that contemporary gene flow is sufficient to maintain genetic connectivity in this rare and restricted Seychelles endemic, in contrast to other island endemic tree species with limited contemporary gene flow.

Genetic Connectivity of the Moth Pollinated Tree Glionnetia sericea in a Highly Fragmented Habitat

PubMed Central

Finger, Aline; Valentin, Terence; Ghazoul, Jaboury

2014-01-01

Long-distance gene flow is thought to be one prerequisite for the persistence of plant species in fragmented environments. Human influences have led to severe fragmentation of native habitats in the Seychelles islands, with many species surviving only in small and isolated populations. The endangered Seychelles endemic tree Glionnetia sericea is restricted to altitudes between 450 m and 900 m where the native forest vegetation has been largely lost and replaced with exotic invasives over the last 200 years. This study explores the genetic and ecological consequences of population fragmentation in this species by analysing patterns of genetic diversity in a sample of adults, juveniles and seeds, and by using controlled pollination experiments. Our results show no decrease in genetic diversity and no increase in genetic structuring from adult to juvenile cohorts. Despite significant inbreeding in some populations, there is no evidence of higher inbreeding in juvenile cohorts relative to adults. A Bayesian structure analysis and a tentative paternity analysis indicate extensive historical and contemporary gene flow among remnant populations. Pollination experiments and a paternity analysis show that Glionnetia sericea is self-compatible. Nevertheless, outcrossing is present with 7% of mating events resulting from pollen transfer between populations. Artificial pollination provided no evidence for pollen limitation in isolated populations. The highly mobile and specialized hawkmoth pollinators (Agrius convolvuli and Cenophodes tamsi; Sphingidae) appear to promote extensive gene flow, thus mitigating the potential negative ecological and genetic effects of habitat fragmentation in this species. We conclude that contemporary gene flow is sufficient to maintain genetic connectivity in this rare and restricted Seychelles endemic, in contrast to other island endemic tree species with limited contemporary gene flow. PMID:25347541

Atmospheric-pressure ionization and fragmentation of peptides by solution-cathode glow discharge† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c6sc02032a Click here for additional data file.

PubMed Central

Schwartz, Andrew J.; Walton, Courtney L.; Williams, Kelsey L.; Hieftje, Gary M.

2016-01-01

Modern “-omics” (e.g., proteomics, glycomics, metabolomics, etc.) analyses rely heavily on electrospray ionization and tandem mass spectrometry to determine the structural identity of target species. Unfortunately, these methods are limited to specialized mass spectrometry instrumentation. Here, a novel approach is described that enables ionization and controlled, tunable fragmentation of peptides at atmospheric pressure. In the new source, a direct-current plasma is sustained between a tapered metal rod and a flowing sample-containing solution. As the liquid stream contacts the electrical discharge, peptides from the solution are volatilized, ionized, and fragmented. At high discharge currents (e.g., 70 mA), electrospray-like spectra are observed, dominated by singly and doubly protonated molecular ions. At lower currents (35 mA), many peptides exhibit extensive fragmentation, with a-, b-, c-, x-, and y-type ion series present as well as complex fragments, such as d-type ions, not previously observed with atmospheric-pressure dissociation. Though the mechanism of fragmentation is currently unclear, observations indicate it could result from the interaction of peptides with gas-phase radicals or ultraviolet radiation generated within the plasma. PMID:28451101

Characterization of limonin glucoside metabolites from human prostate cell culture medium using high-performance liquid chromatography/electrospray ionization mass spectrometry and tandem mass spectrometry.

PubMed

Tian, Qingguo; Kent, Kyle D; Bomser, Joshua A; Schwartz, Steven J

2004-01-01

The metabolism of limonin 17-beta-D-glucopyranoside (LG) by non-cancerous (RWPE-1) and cancerous (PC-3) human prostate epithelial cells was investigated using high-performance liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) with in-source fragmentation and tandem mass spectrometry (MS/MS). During positive ion LC/ESI-MS, LG formed an abundant sodiated species ([M+Na]+) while the protonated molecule was barely observable. [M+Na]+ further fragmented into the less abundant [LARL+H]+ and a predominantly protonated aglycone molecule (limonin) due to in-source fragmentation. The major metabolite, limonin A-ring lactone (LARL), formed an abundant protonated molecule that was fragmented into a protonated molecule of limonin by loss of one molecule of water. In MS/MS by collisionally activated dissociation (CAD), LG produced the sodiated aglycone, [aglycone+Na]+, while LARL fragmented into [M+H]+ of limonin and fragment ions resulted by further loss of water, carbon monoxide and carbon dioxide, indicating the presence of oxygenated-ring structures. The limits of detection of LG were 0.4 and 20 fmol in selected-ion monitoring (SIM) and selected-reaction monitoring (SRM) detection, respectively. Copyright 2004 John Wiley & Sons, Ltd.

High-Efficiency Ligation and Recombination of DNA Fragments by Vertebrate Cells

NASA Astrophysics Data System (ADS)

Miller, Cynthia K.; Temin, Howard M.

1983-05-01

DNA-mediated gene transfer (transfection) is used to introduce specific genes into vertebrate cells. Events soon after transfection were quantitatively analyzed by determining the infectivity of the DNA from an avian retrovirus and of mixtures of subgenomic fragments of this DNA. The limiting step of transfection with two DNA molecules is the uptake by a single cell of both DNA's in a biologically active state. Transfected cells mediate ligation and recombination of physically unlinked DNA's at nearly 100 percent efficiency.

The Heats of Formation of GaCl3 and its Fragments

NASA Technical Reports Server (NTRS)

Bauschlicher, Charles W., Jr.; Arnold, James (Technical Monitor)

1998-01-01

The heats of formation of GaC13 and its fragments are computed. The geometries and frequencies are obtained at the B3LYP level. The CCSD(T) approach is used to solve the correlation problem. The effect of Ga 3d correlation is studied, and found to affect the bond energies by up to 1 kcal/mol. Both basis set extrapolation and bond functions are considered as ways to approach the basis set limit. Spin-orbit and scalar relativistic effects are also considered.

Variational principle model for the nuclear caloric curve

DOE Office of Scientific and Technical Information (OSTI.GOV)

Das Gupta, S.

2005-12-15

Following the lead of a recent work, I perform a variational principle model calculation for the nuclear caloric curve. A Skyrme-type interaction with and without momentum dependence is used. The calculation is done for a large nucleus, i.e., in the nuclear matter limit. Thus I address the issue of volume fragmentation only. Nonetheless, the results are similar to the previous, largely phenomenological calculation for a finite nucleus. I find that the onset of fragmentation can be sudden as a function of temperature or excitation energy.

Analysis methods for Kevlar shield response to rotor fragments

NASA Technical Reports Server (NTRS)

Gerstle, J. H.

1977-01-01

Several empirical and analytical approaches to rotor burst shield sizing are compared and principal differences in metal and fabric dynamic behavior are discussed. The application of transient structural response computer programs to predict Kevlar containment limits is described. For preliminary shield sizing, present analytical methods are useful if insufficient test data for empirical modeling are available. To provide other information useful for engineering design, analytical methods require further developments in material characterization, failure criteria, loads definition, and post-impact fragment trajectory prediction.

Localized extinction of an arboreal desert lizard caused by habitat fragmentation

USGS Publications Warehouse

Munguia-Vega, Adrian; Rodriguez-Estrella, Ricardo; Shaw, William W.; Culver, Melanie

2013-01-01

We adopted a species’ perspective for predicting extinction risk in a small, endemic, and strictly scansorial lizard (Urosaurus nigricaudus), in an old (∼60 year) and highly fragmented (8% habitat remaining) agricultural landscape from the Sonoran Desert, Mexico. We genotyped 10 microsatellite loci in 280 individuals from 11 populations in fragmented and continuous habitat. Individual dispersal was restricted to less than 400 m, according to analyses of spatial autocorrelation and spatially explicit Bayesian assignment methods. Within this scale, continuous areas and narrow washes with native vegetation allowed high levels of gene flow over tens of kilometers. In the absence of the native vegetation, cleared areas and highways were identified as partial barriers. In contrast, outside the scale of dispersal, cleared areas behaved as complete barriers, and surveys corroborated the species went extinct after a few decades in all small (less than 45 ha), isolated habitat fragments. No evidence for significant loss of genetic diversity was found, but results suggested fragmentation increased the spatial scale of movements, relatedness, genetic structure, and potentially affected sex-biased dispersal. A plausible threshold of individual dispersal predicted only 23% of all fragments in the landscape were linked with migration from continuous habitat, while complete barriers isolated the majority of fragments. Our study suggested limited dispersal, coupled with an inability to use a homogeneous and hostile matrix without vegetation and shade, could result in frequent time-delayed extinctions of small ectotherms in highly fragmented desert landscapes, particularly considering an increase in the risk of overheating and a decrease in dispersal potential induced by global warming.

AFEAP cloning: a precise and efficient method for large DNA sequence assembly.

PubMed

Zeng, Fanli; Zang, Jinping; Zhang, Suhua; Hao, Zhimin; Dong, Jingao; Lin, Yibin

2017-11-14

Recent development of DNA assembly technologies has spurred myriad advances in synthetic biology, but new tools are always required for complicated scenarios. Here, we have developed an alternative DNA assembly method named AFEAP cloning (Assembly of Fragment Ends After PCR), which allows scarless, modular, and reliable construction of biological pathways and circuits from basic genetic parts. The AFEAP method requires two-round of PCRs followed by ligation of the sticky ends of DNA fragments. The first PCR yields linear DNA fragments and is followed by a second asymmetric (one primer) PCR and subsequent annealing that inserts overlapping overhangs at both sides of each DNA fragment. The overlapping overhangs of the neighboring DNA fragments annealed and the nick was sealed by T4 DNA ligase, followed by bacterial transformation to yield the desired plasmids. We characterized the capability and limitations of new developed AFEAP cloning and demonstrated its application to assemble DNA with varying scenarios. Under the optimized conditions, AFEAP cloning allows assembly of an 8 kb plasmid from 1-13 fragments with high accuracy (between 80 and 100%), and 8.0, 11.6, 19.6, 28, and 35.6 kb plasmids from five fragments at 91.67, 91.67, 88.33, 86.33, and 81.67% fidelity, respectively. AFEAP cloning also is capable to construct bacterial artificial chromosome (BAC, 200 kb) with a fidelity of 46.7%. AFEAP cloning provides a powerful, efficient, seamless, and sequence-independent DNA assembly tool for multiple fragments up to 13 and large DNA up to 200 kb that expands synthetic biologist's toolbox.

Oriented Immobilization of Fab Fragments by Site-Specific Biotinylation at the Conserved Nucleotide Binding Site for Enhanced Antigen Detection.

PubMed

Mustafaoglu, Nur; Alves, Nathan J; Bilgicer, Basar

2015-09-08

Oriented immobilization of antibodies and antibody fragments has become increasingly important as a result of the efforts to reduce the size of diagnostic and sensor devices to miniaturized dimensions for improved accessibility to the end-user. Reduced dimensions of sensor devices necessitate the immobilized antibodies to conserve their antigen binding activity for proper operation. Fab fragments are becoming more commonly used in small-scaled diagnostic devices due to their small size and ease of manufacture. In this study, we used the previously described UV-NBS(Biotin) method to functionalize Fab fragments with IBA-EG11-Biotin linker utilizing UV energy to initiate a photo-cross-linking reaction between the nucleotide binding site (NBS) on the Fab fragment and IBA-Biotin molecule. Our results demonstrate that immobilization of biotinylated Fab fragments via UV-NBS(Biotin) method generated the highest level of immobilized Fab on surfaces when compared to other typical immobilization methods while preserving antigen binding activity. UV-NBS(Biotin) method provided 432-fold, 114-fold, and 29-fold improved antigen detection sensitivity than physical adsorption, NHS-Biotin, and ε-NH3(+), methods, respectively. Additionally, the limit of detection (LOD) for PSA utilizing Fab fragments immobilized via UV-NBS(Biotin) method was significantly lower than that of the other immobilization methods, with an LOD of 0.4 pM PSA. In summary, site-specific biotinylation of Fab fragments without structural damage or loss in antigen binding activity provides a wide range of application potential for UV-NBS immobilization technique across numerous diagnostic devices and nanotechnologies.

Rhizome Fragmentation by Vertical Disks Reduces Elymus repens Growth and Benefits Italian Ryegrass-White Clover Crops

PubMed Central

Ringselle, Björn; Bertholtz, Erik; Magnuski, Ewa; Brandsæter, Lars Olav; Mangerud, Kjell; Bergkvist, Göran

2018-01-01

Tillage controls perennial weeds, such as Elymus repens, partly because it fragments their underground storage organs. However, tillage is difficult to combine with a growing crop, which limits its application. The aim of this study was to evaluate how soil vertical cutting with minimum soil disturbance and mowing affect the growth and competitive ability of E. repens in a grass–clover crop. A tractor-drawn prototype with vertical disks was used to fragment E. repens rhizomes with minimal soil and crop disturbance. In experiments performed in 2014 and 2015 at a field site close to Uppsala, Sweden, the rhizomes were fragmented before crop sowing (ERF), during crop growth (LRF), or both (ERF+LRF). Fragmentation was combined with repeated mowing (yes/no) and four companion crop treatments (none, Italian ryegrass, white clover, and grass/clover mixture). The results showed that in the grass–clover crop, rhizome fragmentation reduced E. repens rhizome biomass production and increased Italian ryegrass shoot biomass. ERF and LRF both reduced E. repens rhizome biomass by about 38% compared with the control, while ERF+LRF reduced it by 63%. Italian ryegrass shoot biomass was increased by 78% by ERF, 170% by LRF and 200% by ERF+LRF. Repeated mowing throughout the experiment reduced E. repens rhizome biomass by about 75%. Combining repeated mowing with rhizome fragmentation did not significantly increase the control effect compared to mowing alone. We concluded that rhizome fragmentation using vertical disks can be used both before sowing and during crop growth to enhance the controlling effect of grass–clover crops on E. repens. PMID:29375611

Lattice enumeration for inverse molecular design using the signature descriptor.

PubMed

Martin, Shawn

2012-07-23

We describe an inverse quantitative structure-activity relationship (QSAR) framework developed for the design of molecular structures with desired properties. This framework uses chemical fragments encoded with a molecular descriptor known as a signature. It solves a system of linear constrained Diophantine equations to reorganize the fragments into novel molecular structures. The method has been previously applied to problems in drug and materials design but has inherent computational limitations due to the necessity of solving the Diophantine constraints. We propose a new approach to overcome these limitations using the Fincke-Pohst algorithm for lattice enumeration. We benchmark the new approach against previous results on LFA-1/ICAM-1 inhibitory peptides, linear homopolymers, and hydrofluoroether foam blowing agents. Software implementing the new approach is available at www.cs.otago.ac.nz/homepages/smartin.

Longitudinal study of sperm DNA fragmentation as measured by terminal uridine nick end-labelling assay.

PubMed

Sergerie, M; Laforest, G; Boulanger, K; Bissonnette, F; Bleau, G

2005-07-01

One major limitation in the use of sperm DNA fragmentation as measured by the TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) assay is the paucity of solid data on the stability of this parameter. The objective of our study was to evaluate variations in the degree of sperm DNA fragmentation, as measured by the TUNEL assay, over a 6 month period. Five donors provided semen samples (total 107) on the average three times per month, and 10 infertility patients provided semen samples every 4 weeks (total 58). The mean percentage of sperm DNA fragmentation for donors was 13.18%, the within-donor standard deviation (SD(W) = 3.79%) was small compared to between-donor (SD(B) = 17.56%). For the group of patients, the mean percentage of sperm DNA fragmentation was 22.44%, with SD(W) of 4.43% within patients and SD(B) of 29.48% between patients. No seasonal rhythm was observed during the study. The intra-class correlation coefficient for all subjects combined was 0.83. Compared to sperm concentration, individual coefficients of variation for sperm DNA fragmentation indicated less variability in four subjects, but were similar in the others. This longitudinal study shows that sperm DNA fragmentation is a parameter with good stability (repeatability) over time; it can be taken as a baseline both in healthy fertile men and in patients from infertility couples.

Tropical Forest Fragmentation Limits Movements, but Not Occurrence of a Generalist Pollinator Species

PubMed Central

Volpe, Noelia L.; Robinson, W. Douglas; Frey, Sarah J. K.; Hadley, Adam S.; Betts, Matthew G.

2016-01-01

Habitat loss and fragmentation influence species distributions and therefore ecological processes that depend upon them. Pollination may be particularly susceptible to fragmentation, as it depends on frequent pollinator movement. Unfortunately, most pollinators are too small to track efficiently which has precluded testing the hypothesis that habitat fragmentation reduces or eliminates pollen flow by disrupting pollinator movement. We used radio-telemetry to examine space use of the green hermit hummingbird (Phaethornis guy), an important ‘hub’ pollinator of understory flowering plants across substantial portions of the neotropics and the primary pollinator of a keystone plant which shows reduced pollination success in fragmented landscapes. We found that green hermits strongly avoided crossing large stretches of non-forested matrix and preferred to move along stream corridors. Forest gaps as small as 50 m diminished the odds of movement by 50%. Green hermits occurred almost exclusively inside the forest, with the odds of occurrence being 8 times higher at points with >95% canopy cover compared with points having <5% canopy cover. Nevertheless, surprisingly. the species occurred in fragmented landscapes with low amounts of forest (~30% within a 2 km radius). Our results indicate that although green hermits are present even in landscapes with low amounts of tropical forest, movement within these landscapes ends up strongly constrained by forest gaps. Restricted movement of pollinators may be an underappreciated mechanism for widespread declines in pollination and plant fitness in fragmented landscapes, even when in the presence of appropriate pollinators. PMID:27941984

Nudged elastic band method and density functional theory calculation for finding a local minimum energy pathway of p-benzoquinone and phenol fragmentation in mass spectrometry.

PubMed

Sugimura, Natsuhiko; Igarashi, Yoko; Aoyama, Reiko; Shibue, Toshimichi

2017-02-01

Analysis of the fragmentation pathways of molecules in mass spectrometry gives a fundamental insight into gas-phase ion chemistry. However, the conventional intrinsic reaction coordinates method requires knowledge of the transition states of ion structures in the fragmentation pathways. Herein, we use the nudged elastic band method, using only the initial and final state ion structures in the fragmentation pathways, and report the advantages and limitations of the method. We found a minimum energy path of p-benzoquinone ion fragmentation with two saddle points and one intermediate structure. The primary energy barrier, which corresponded to the cleavage of the C-C bond adjacent to the CO group, was calculated to be 1.50 eV. An additional energy barrier, which corresponded to the cleavage of the CO group, was calculated to be 0.68 eV. We also found an energy barrier of 3.00 eV, which was the rate determining step of the keto-enol tautomerization in CO elimination from the molecular ion of phenol. The nudged elastic band method allowed the determination of a minimum energy path using only the initial and final state ion structures in the fragmentation pathways, and it provided faster than the conventional intrinsic reaction coordinates method. In addition, this method was found to be effective in the analysis of the charge structures of the molecules during the fragmentation in mass spectrometry.

Fragmentation of Massive Dense Cores Down to <~ 1000 AU: Relation between Fragmentation and Density Structure

NASA Astrophysics Data System (ADS)

Palau, Aina; Estalella, Robert; Girart, Josep M.; Fuente, Asunción; Fontani, Francesco; Commerçon, Benoit; Busquet, Gemma; Bontemps, Sylvain; Sánchez-Monge, Álvaro; Zapata, Luis A.; Zhang, Qizhou; Hennebelle, Patrick; di Francesco, James

2014-04-01

In order to shed light on the main physical processes controlling fragmentation of massive dense cores, we present a uniform study of the density structure of 19 massive dense cores, selected to be at similar evolutionary stages, for which their relative fragmentation level was assessed in a previous work. We inferred the density structure of the 19 cores through a simultaneous fit of the radial intensity profiles at 450 and 850 μm (or 1.2 mm in two cases) and the spectral energy distribution, assuming spherical symmetry and that the density and temperature of the cores decrease with radius following power-laws. Even though the estimated fragmentation level is strictly speaking a lower limit, its relative value is significant and several trends could be explored with our data. We find a weak (inverse) trend of fragmentation level and density power-law index, with steeper density profiles tending to show lower fragmentation, and vice versa. In addition, we find a trend of fragmentation increasing with density within a given radius, which arises from a combination of flat density profile and high central density and is consistent with Jeans fragmentation. We considered the effects of rotational-to-gravitational energy ratio, non-thermal velocity dispersion, and turbulence mode on the density structure of the cores, and found that compressive turbulence seems to yield higher central densities. Finally, a possible explanation for the origin of cores with concentrated density profiles, which are the cores showing no fragmentation, could be related with a strong magnetic field, consistent with the outcome of radiation magnetohydrodynamic simulations. The James Clerk Maxwell Telescope is operated by the Joint Astronomy Centre on behalf of the Science and Technology Facilities Council of the United Kingdom, the Netherlands Organisation for Scientific Research, and the National Research Council of Canada.

Time-resolved ion imaging at free-electron lasers using TimepixCam

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fisher-Levine, Merlin; Boll, Rebecca; Ziaee, Farzaneh

In this paper, the application of a novel fast optical-imaging camera, TimepixCam, to molecular photoionization experiments using the velocity-map imaging technique at a free-electron laser is described. TimepixCam is a 256 × 256 pixel CMOS camera that is able to detect and time-stamp ion hits with 20 ns timing resolution, thus making it possible to record ion momentum images for all fragment ions simultaneously and avoiding the need to gate the detector on a single fragment. This allows the recording of significantly more data within a given amount of beam time and is particularly useful for pump–probe experiments, where drifts,more » for example, in the timing and pulse energy of the free-electron laser, severely limit the comparability of pump–probe scans for different fragments taken consecutively. Finally, in principle, this also allows ion–ion covariance or coincidence techniques to be applied to determine angular correlations between fragments.« less

Feedback between intracellular flow, signaling and active stresses in Physarum plasmodial fragments

NASA Astrophysics Data System (ADS)

Zhang, Shun; Guy, Robert; Del Alamo, Juan Carlos

2016-11-01

Physarum polycephalum is a multinucleated slime mold whose endoplasm flows periodically driven by the contraction of its ectoplasm, a dense shell of F-actin cross-linked by myosin molecular motors and attached to the cell membrane. Ectoplasm contractions are regulated by calcium ions whose propagation is in turn governed by the flow. We study experimentally how this feedback leads to auto-oscillation by simultaneously measuring endoplasmic flow speed and rheological properties, the traction stresses between the ectoplasm and its substratum and the distribution of endoplasmic free calcium ions. We find that physarum fragments smaller than 100 microns remain round and stay in place. However, larger fragments break symmetry leading to sustained forward locomotion, in process that is reminiscent of an interfacial instability that seems to settle around two different limit cycles (traveling waves and standing waves). By using different adhesive coatings in the substratum we investigate the role of substratum friction in the emergence of coherent endoplasmic flow patterns and overall physarum fragment locomotion.

Time-resolved ion imaging at free-electron lasers using TimepixCam

DOE PAGES

Fisher-Levine, Merlin; Boll, Rebecca; Ziaee, Farzaneh; ...

2018-02-20

In this paper, the application of a novel fast optical-imaging camera, TimepixCam, to molecular photoionization experiments using the velocity-map imaging technique at a free-electron laser is described. TimepixCam is a 256 × 256 pixel CMOS camera that is able to detect and time-stamp ion hits with 20 ns timing resolution, thus making it possible to record ion momentum images for all fragment ions simultaneously and avoiding the need to gate the detector on a single fragment. This allows the recording of significantly more data within a given amount of beam time and is particularly useful for pump–probe experiments, where drifts,more » for example, in the timing and pulse energy of the free-electron laser, severely limit the comparability of pump–probe scans for different fragments taken consecutively. Finally, in principle, this also allows ion–ion covariance or coincidence techniques to be applied to determine angular correlations between fragments.« less

Ligament Mediated Fragmentation of Viscoelastic Liquids

NASA Astrophysics Data System (ADS)

Keshavarz, Bavand; Houze, Eric C.; Moore, John R.; Koerner, Michael R.; McKinley, Gareth H.

2016-10-01

The breakup and atomization of complex fluids can be markedly different than the analogous processes in a simple Newtonian fluid. Atomization of paint, combustion of fuels containing antimisting agents, as well as physiological processes such as sneezing are common examples in which the atomized liquid contains synthetic or biological macromolecules that result in viscoelastic fluid characteristics. Here, we investigate the ligament-mediated fragmentation dynamics of viscoelastic fluids in three different canonical flows. The size distributions measured in each viscoelastic fragmentation process show a systematic broadening from the Newtonian solvent. In each case, the droplet sizes are well described by Gamma distributions which correspond to a fragmentation-coalescence scenario. We use a prototypical axial step strain experiment together with high-speed video imaging to show that this broadening results from the pronounced change in the corrugated shape of viscoelastic ligaments as they separate from the liquid core. These corrugations saturate in amplitude and the measured distributions for viscoelastic liquids in each process are given by a universal probability density function, corresponding to a Gamma distribution with nmin=4 . The breadth of this size distribution for viscoelastic filaments is shown to be constrained by a geometrical limit which can not be exceeded in ligament-mediated fragmentation phenomena.

The RNA methyltransferase Dnmt2 is required for efficient Dicer-2-dependent siRNA pathway activity in Drosophila.

PubMed

Durdevic, Zeljko; Mobin, Mehrpouya Balaghy; Hanna, Katharina; Lyko, Frank; Schaefer, Matthias

2013-09-12

Transfer RNA (tRNA) fragmentation in response to stress conditions has been described in many organisms. tRNA fragments have been found in association with small interfering RNA (siRNA) components, but the biological role of these interactions remains unclear. We report here that the tRNA methyltransferase Dnmt2 is essential for efficient Dicer-2 (Dcr-2) function in Drosophila. Using small RNA (sRNA) sequencing, we confirmed that Dnmt2 limits the extent of tRNA fragmentation during the heat-shock response. tRNAs as well as tRNA fragments serve as Dcr-2 substrates, and Dcr-2 degrades tRNA-derived sequences, especially under heat-shock conditions. tRNA-derived RNAs are able to inhibit Dcr-2 activity on long double-stranded RNAs (dsRNAs). Consequently, heat-shocked Dnmt2 mutant animals accumulate dsRNAs, produce fewer siRNAs, and show misregulation of siRNA pathway-dependent genes. These results reveal the impact of tRNA fragmentation on siRNA pathways and implicate tRNA modifications in the regulation of sRNA homeostasis during the heat-shock response. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

Ligament Mediated Fragmentation of Viscoelastic Liquids.

PubMed

Keshavarz, Bavand; Houze, Eric C; Moore, John R; Koerner, Michael R; McKinley, Gareth H

2016-10-07

The breakup and atomization of complex fluids can be markedly different than the analogous processes in a simple Newtonian fluid. Atomization of paint, combustion of fuels containing antimisting agents, as well as physiological processes such as sneezing are common examples in which the atomized liquid contains synthetic or biological macromolecules that result in viscoelastic fluid characteristics. Here, we investigate the ligament-mediated fragmentation dynamics of viscoelastic fluids in three different canonical flows. The size distributions measured in each viscoelastic fragmentation process show a systematic broadening from the Newtonian solvent. In each case, the droplet sizes are well described by Gamma distributions which correspond to a fragmentation-coalescence scenario. We use a prototypical axial step strain experiment together with high-speed video imaging to show that this broadening results from the pronounced change in the corrugated shape of viscoelastic ligaments as they separate from the liquid core. These corrugations saturate in amplitude and the measured distributions for viscoelastic liquids in each process are given by a universal probability density function, corresponding to a Gamma distribution with n_{min}=4. The breadth of this size distribution for viscoelastic filaments is shown to be constrained by a geometrical limit which can not be exceeded in ligament-mediated fragmentation phenomena.

Effectiveness of Alpha-toxin Fab Monoclonal Antibody Therapy in Limiting the Pathology of Staphylococcus aureus Keratitis.

PubMed

Caballero, Armando R; Foletti, Davide L; Bierdeman, Michael A; Tang, Aihua; Arana, Angela M; Hasa-Moreno, Adela; Sangalang, Emma Ruth B; O'Callaghan, Richard J

2015-08-01

To investigate the effectiveness of a high-affinity human monoclonal antibody Fab fragment to Staphylococcus aureus alpha-toxin (LTM14 Fab) as therapy for S. aureus keratitis. A single topical drop of the LTM14 Fab antibody to alpha-toxin alone, or in 0.006% benzalkonium chloride (BAK), was applied every 30 min to S. aureus-infected rabbit corneas from 9 to 14 hours post-infection. Erosions and pathology were measured at 15 h post-infection. LTM14 Fab with BAK limited corneal erosions better than LTM14 Fab alone (p = 0.036), and both limited erosions compared to untreated eyes (p ≤ 0.0001). Overall pathology was similar in all groups (p ≥ 0.070), but iritis and chemosis were reduced by treatment (p ≤ 0.036). The high-affinity human monoclonal Fab fragment antibody (LTM14 Fab) to S. aureus alpha-toxin was effective in reducing corneal damage during S. aureus keratitis.

Interrami intraoral fixation technique for severe mandibular rifle fragmented bullet injury management.

PubMed

Shuker, Sabri T

2013-07-01

Interrami intraoral Kirschner wire fixation technique is presented for the reduction, stabilization, and immobilization of a pulverized and avulsed lower jaw caused by rifle fragmented bullet injuries. This indirect mandibular war injury fixation technique was tolerated by the patients and tissue more than any indirect external fixation. In addition, it is easier than open reduction using large bone plates for disrupted ballistics mandibular injury defects. An interrami intraoral fixation is appropriate for severely disrupted mandibular hard and soft tissues, and has been adapted in cases of mass casualties and limited resources. Benefits of use include limited hospital beds and fewer follow-up visits. Rifle fragmented bullet injuries need more attention for several reasons: not only because of the higher mortality and devastating nature of the injuries, but also because these injuries are responsible for an unreported type of bullet biomechanism wounding in the craniofacial region. In turn, this necessitates specialized victim management. The survival rates depend on immediate proper execution of airway, breathing, and circulation, which become more complicated as it relates to airway compromise and oropharyngeal hemorrhage resuscitation. Survival is predicated on the implementation of feasible, sensible, life-saving techniques that are applied at the appropriate time.

Discrete RNA libraries from pseudo-torsional space

PubMed Central

Humphris-Narayanan, Elisabeth

2012-01-01

The discovery that RNA molecules can fold into complex structures and carry out diverse cellular roles has led to interest in developing tools for modeling RNA tertiary structure. While significant progress has been made in establishing that the RNA backbone is rotameric, few libraries of discrete conformations specifically for use in RNA modeling have been validated. Here, we present six libraries of discrete RNA conformations based on a simplified pseudo-torsional notation of the RNA backbone, comparable to phi and psi in the protein backbone. We evaluate the ability of each library to represent single nucleotide backbone conformations and we show how individual library fragments can be assembled into dinucleotides that are consistent with established RNA backbone descriptors spanning from sugar to sugar. We then use each library to build all-atom models of 20 test folds and we show how the composition of a fragment library can limit model quality. Despite the limitations inherent in using discretized libraries, we find that several hundred discrete fragments can rebuild RNA folds up to 174 nucleotides in length with atomic-level accuracy (<1.5Å RMSD). We anticipate the libraries presented here could easily be incorporated into RNA structural modeling, analysis, or refinement tools. PMID:22425640

Genetic erosion impedes adaptive responses to stressful environments

PubMed Central

Bijlsma, R; Loeschcke, Volker

2012-01-01

Biodiversity is increasingly subjected to human-induced changes of the environment. To persist, populations continually have to adapt to these often stressful changes including pollution and climate change. Genetic erosion in small populations, owing to fragmentation of natural habitats, is expected to obstruct such adaptive responses: (i) genetic drift will cause a decrease in the level of adaptive genetic variation, thereby limiting evolutionary responses; (ii) inbreeding and the concomitant inbreeding depression will reduce individual fitness and, consequently, the tolerance of populations to environmental stress. Importantly, inbreeding generally increases the sensitivity of a population to stress, thereby increasing the amount of inbreeding depression. As adaptation to stress is most often accompanied by increased mortality (cost of selection), the increase in the ‘cost of inbreeding’ under stress is expected to severely hamper evolutionary adaptive processes. Inbreeding thus plays a pivotal role in this process and is expected to limit the probability of genetically eroded populations to successfully adapt to stressful environmental conditions. Consequently, the dynamics of small fragmented populations may differ considerably from large nonfragmented populations. The resilience of fragmented populations to changing and deteriorating environments is expected to be greatly decreased. Alleviating inbreeding depression, therefore, is crucial to ensure population persistence. PMID:25568035

Effects of introduction of new resources and fragmentation of existing resources on limiting wealth distribution in asset exchange models

NASA Astrophysics Data System (ADS)

Ali Saif, M.; Gade, Prashant M.

2009-03-01

Pareto law, which states that wealth distribution in societies has a power-law tail, has been the subject of intensive investigations in the statistical physics community. Several models have been employed to explain this behavior. However, most of the agent based models assume the conservation of number of agents and wealth. Both these assumptions are unrealistic. In this paper, we study the limiting wealth distribution when one or both of these assumptions are not valid. Given the universality of the law, we have tried to study the wealth distribution from the asset exchange models point of view. We consider models in which (a) new agents enter the market at a constant rate (b) richer agents fragment with higher probability introducing newer agents in the system (c) both fragmentation and entry of new agents is taking place. While models (a) and (c) do not conserve total wealth or number of agents, model (b) conserves total wealth. All these models lead to a power-law tail in the wealth distribution pointing to the possibility that more generalized asset exchange models could help us to explain the emergence of a power-law tail in wealth distribution.

Study of Projectile Fragmentation Reactions at Intermediate Energies

NASA Astrophysics Data System (ADS)

Souliotis, George A.

The experimental measurements of the present work represent a first attempt towards obtaining systematic data of projectile fragment distributions near 0 degrees produced by intermediate-energy beams delivered by the K1200 cyclotron at the NSCL. The measurements were performed with two different devices operated as 0-degree spectrometers: the K1200 interim beamline and the A1200 mass separator. Momentum distributions and limited angular distributions of projectile -like fragments (PLFs) from a ^{14} N (75 MeV/u) beam with ^{27 }Al and ^{181}Ta targets were measured using the interim K1200 beamline, but the results were of limited quality. In a first series of measurements with the A1200, the 0-degree momentum distributions of PLFs from several beams (^{20} Ne (85 MeV/u), ^{15}N (70 MeV/u) and ^{18}O (70 Mev/u)) were measured, but only a limited number of complete momentum spectra were obtained. In a second series, a detailed measurement of neutron-rich PLFs from the reaction ^{18}O (80 MeV/u) + ^{27}Al was successfully performed. Accurate measurements of 0-degree momentum distributions and relative yields of PLFs were obtained and analysed. Analysis of the centroids of the momentum-per -nucleon (velocity) distributions of the fragments indicated that, along with the fragmentation (break-up) channels, direct stripping and/or pick-up channels contribute significantly to the production of the fragments. Interestingly, the mean velocities of nucleon pick-up products were considerably lower than that of the beam, consistent with momentum conservation and the additional assumption that the nucleon(s) picked -up from the target have preferentially momenta (on average) equal to the nuclear Fermi momentum oriented in the direction of the projectile motion. The widths of the momentum distributions of both nucleon-removal and nucleon pick-up products were in agreement with the independent-particle model of Goldhaber and the cluster stripping model of Friedman. In the case of pick-up products, the momentum distributions of both the projectile part and the picked-up nucleon(s) had to be taken into account. Calculations of yields with the abrasion-ablation model and the intranuclear cascade model showed that both these high-energy models were not able to describe the trend of the data. However, calculations with Friedman's model provided a good description of the experimental yields, implying that a break-up (and/or pick -up) mechanism involving low excitation energy is responsible for the production of PLFs from the present light projectile.

An Attempt to Observe Debris from the Breakup of a Titan 3C-4 Transtage

NASA Technical Reports Server (NTRS)

Barker, E. S.; Matney, M. J.; Yanagisawa, T.; Liou, J.-C.; Abercromby, K. J.; Rodriquez, H. M.; Horstman, M. F.; Seitzer, P.

2007-01-01

In February 2007 dedicated observations were made of the orbital space predicted to contain debris from the breakup of the Titan 3C-4 transtage back on February 21, 1992. These observations were carried out on the Michigan Orbital DEbris Survey Telescope (MODEST) in Chile with its 1.3deg field of view. The search region or orbital space (inclination and right ascension of the ascending node (RAAN) was predicted using NASA#s LEGEND (LEO-to-GEO Environment Debris) code to generate a Titan debris cloud. Breakup fragments are created based on the NASA Standard Breakup Model (including fragment size, area-to-mass (A/M), and delta-V distributions). Once fragments are created, they are propagated forward in time with a subroutine GEOPROP. Perturbations included in GEOPROP are those due to solar/lunar gravity, radiation pressure, and major geopotential terms. Barker, et. al, (AMOS Conference Proceedings, 2006, pp. 596-604) used similar LEGEND predictions to correlate survey observations made by MODEST (February 2002) and found several possible night-to-night correlations in the limited survey dataset. One conc lusion of the survey search was to dedicate a MODEST run to observing a GEO region predicted to contain debris fragments and actual Titan debris objects (SSN 25000, 25001 and 30000). Such a dedicated run was undertaken with MODEST between February 17 and 23, 2007 (UT dates). MODEST#s limiting magnitude of 18.0 (S\\N approx.10) corresponds to a size of 22cm assuming a diffuse Lambertian albedo of 0.2. However, based on observed break-up data, we expect most debris fragments to be smaller than 22cm which implies a need to increase the effective sensitivity of MODEST for smaller objects. MODEST#s limiting size can be lowered by increasing the exposure time (20 instead of 5 seconds) and applying special image processing. The special processing combines individual CCD images to detect faint objects that are invisible on a single CCD image. Sub-images are cropped from six consecutive CCD images with pixel shifts between images being consistent with the predicted movement of a Titan object. A median image of all the sub-images is then created leaving only those objects with the proper Titan motion. Limiting the median image in this manner brings the needed computer time to process all images taken on one night down to about 50 hours of CPU time.

How well can step-off and gap distances be reduced when treating intra-articular distal radius fractures with fragment specific fixation when using fluoroscopy.

PubMed

Thiart, M; Ikram, A; Lamberts, R P

2016-12-01

Although fragment specific fixation has proved to be an effective treatment regime, it has not been established how successfully this treatment could be performed using fluoroscopy and what the added value of arthroscopy could be. Establish gap and step-off distances after in intra-articular distal radius fractures that have been treated with fragment specific fixation while using fluoroscopy. Forty-four patients with an intra-articular distal radius fracture were treated with fragment specific fixation while using fluoroscopy. After the treatment of the intra-articular distal radius fracture with fragment specific fixation and the use of fluoroscopy, but before the completion of the surgical intervention, all gap, and step-off distances were determined by using arthroscopy. In addition, the joint was checked for any other wrist pathologies. Arthroscopy after the surgical intervention showed that in 37 patients no gap distances could be detected, while in six patients a gap distance of≤2mm was found and in one patient, a gap distance of 3mm. Similarly, arthroscopy revealed no step-off distances in 33 patients, while in 11 patients a step-off distance of≤2mm was found. Although additional wrist pathologies were found in 48% of our population, only one patient needed surgical intervention. Three months after the surgical intervention wrist flexion was 41±10°, wrist extension 51±17°, ulnar deviation 19±10°, radial deviation 32±12° while patients could pronate and supinate their wrist to 85±5° and 74±20°, respectively. Intra-articular distal radius fractures can be treated successfully with fragment specific fixation and the use of fluoroscopy. As almost all gap and step-off distances could be reduced to an acceptable level, the scope for arthroscopy to further improve this treatment regime is limited. The functional outcome scores that were found 3 months after the surgical intervention were similar to what has been reported in other studies using different treatment option. These findings suggest that fragment specific fixation is a good alternative for treating intra-articular distal radius fractures. As in most cases, only fluoroscopy is needed for fragment specific fixation, this treatment technique is a good treatment option for resource-limited hospitals, setting who do not have access to arthroscopy. III, case-control study. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Hubble Space Telescope Observations of the Nucleus Fragment 73P/Schwassmann-Wachmann 3-B

NASA Astrophysics Data System (ADS)

Toth, I.; Lamy, P. L.; Weaver, H. A.

2003-05-01

The nucleus of 73P/Schwassmann-Wachmann 3 (73P/SW3), a Jupiter-Family comet, has non-tidally broken into at least three components A, B and C, in autumn 1995. Fragment B was detected with the Planetary Camera 2 of the Hubble Space Telescope on 26 November 2001. i.e., after the comet's perihelion passage on 27.9 January 2001, when it was at 3.25 AU from the Sun, 2.33 AU from the Earth, and at a solar phase angle of 7.4o. The high spatial resolution of the PC2 allowed to separate the signal of the faint nucleus fragment from that of its active coma, and we measured its R magnitude from our images taken with the F675W filter. Assuming a spherical body whose geometric albedo is 0.04 and a linear phase coefficient of 0.04 mag/deg for the R band, we derived an effective radius of 0.68+/-0.04 km. The short time span of our HST observations did not allow to determine its shape but the lower limit of its axial ratio a/b (assuming a prolate spheroid) is ˜1.16. From the pre-breakup radius of the original nucleus of 1.3 km determined by Boehnhardt et al. (2002: EMPl 90, 131), we found that the fractional volume of fragment B is about 14%. Its size of a few hundred meters, typical of fragments of other split comets, is probably too large for a primordial building block and indicate that it still is a conglomerate; further disintegration of this fragment remains possible. Its level of activity is remarkably high in spite of a heliocentric distance of ˜3.2 AU, just beyond the limit for sublimation of water ice (2.8 AU). We measured an Afρ of 19.6-23.2 cm but the determination of a dust production rate is precluded by the absence of gas production rates at the time of our observation. Ground-based observations performed in 2001 by Boehnhardt et al. (2002: EMPl 90, 131) indicate that another fragment (E) has already disappeared, and we conclude that this may well be short-term fate of fragment B as it may not survive its next perihelion passage in 2006. The tau-Herculids meteor swarm associated to 73P/SW3 will likely experience an increase of activity. This work was supported by the CNRS, CNES, and Université de Provence, France, and by the Hungarian Academy of Science through grant No. 9871.

Cytosolic expression of functional Fab fragments in Escherichia coli using a novel combination of dual SUMO expression cassette and EnBase® cultivation mode.

PubMed

Rezaie, F; Davami, F; Mansouri, K; Agha Amiri, S; Fazel, R; Mahdian, R; Davoudi, N; Enayati, S; Azizi, M; Khalaj, V

2017-05-08

The Escherichia coli expression system is highly effective in producing recombinant proteins. However, there are some limitations in this system, especially in obtaining correctly folded forms of some complex proteins such as Fab fragments. To improve the solubility and folding quality of Fab fragments, we have examined the effect of simultaneous application of a SUMO fusion tag, EnBase ® cultivation mode and a redox mutant strain in the E. coli expression system. A bicistronic gene construct was designed to express an antivascular endothelial growth factor (VEGF) Fab fragment as a model system. The construct contained a dual SUMO fusion gene fragment to encode SUMO-tagged heavy and light chains. While the expression of the construct in batch cultures of BL21 or SHuffle ® transformants produced insoluble and unfolded products, the induction of the transformants in EnBase ® medium resulted in soluble and correctly folded Fab fragment, reaching as high as 19% of the total protein in shuffle strain. The functional assays indicated that the biological activity of the target Fab is similar to the commercial anti-VEGF, Lucentis ® . This study demonstrated that the combination of SUMO fusion technology, EnBase ® cultivation system and recruiting a redox mutant of E. coli can efficiently enhance the solubility and productivity of recombinant Fab fragments. The presented strategy provides not only a novel method to produce soluble and active form of an anti-VEGF Fab but also may use in the efficient production of other antibody fragments. © 2017 The Society for Applied Microbiology.

Hyperthermal (1-100 eV) nitrogen ion scattering damage to D-ribose and 2-deoxy-D-ribose films.

PubMed

Deng, Zongwu; Bald, Ilko; Illenberger, Eugen; Huels, Michael A

2007-10-14

Highly charged heavy ion traversal of a biological medium can produce energetic secondary fragment ions. These fragment ions can in turn cause collisional and reactive scattering damage to DNA. Here we report hyperthermal (1-100 eV) scattering of one such fragment ion (N(+)) from biologically relevant sugar molecules D-ribose and 2-deoxy-D-ribose condensed on polycrystalline Pt substrate. The results indicate that N(+) ion scattering at kinetic energies down to 10 eV induces effective decomposition of both sugar molecules and leads to the desorption of abundant cation and anion fragments. Use of isotope-labeled molecules (5-(13)C D-ribose and 1-D D-ribose) partly reveals some site specificity of the fragment origin. Several scattering reactions are also observed. Both ionic and neutral nitrogen atoms abstract carbon from the molecules to form CN(-) anion at energies down to approximately 5 eV. N(+) ions also abstract hydrogen from hydroxyl groups of the molecules to form NH(-) and NH(2) (-) anions. A fraction of OO(-) fragments abstract hydrogen to form OH(-). The formation of H(3)O(+) ions also involves hydrogen abstraction as well as intramolecular proton transfer. These findings suggest a variety of severe damaging pathways to DNA molecules which occur on the picosecond time scale following heavy ion irradiation of a cell, and prior to the late diffusion-limited homogeneous chemical processes.

Synthesis of a posterior indicator protein in normal embryos and double abdomens of Smittia sp. (Chironomidae, Diptera).

PubMed Central

Jäckle, H; Kalthoff, K

1980-01-01

In embryos of the chironomid midge Smittia, synthesis of a posterior indicator protein designated PI1 (Mr approximately 50,000; pI approximately 5.5) forecasts development of an abdomen as opposed to head and thorax. The protein is synthesized several hours before germ anlage formation. In normal embryos at early blastoderm stages, synthesis of PI1 is restricted to posterior embryonic fragments but not to pole cells. In "double-abdomen" embryos, a mirror-image duplication of the abdomen is formed by cells that would otherwise develop into head and thorax. Embryos were programmed for double-abdomen development by UV irradiation of the anterior pole, and half of them were reprogrammed for normal development by subsequent exposure to visible light (photoreversal). Correspondingly, PI1 was synthesized in anterior fragments of UV-irradiated embryos but not after photoreversal. In a control experiment, UV irradiation of the posterior pole caused neither double-abdomen formation nor PI1 synthesis in anterior fragments. The identity of PI1 formed in anterior fragments of prospective double abdomens with the protein found in posterior fragments was revealed by two-dimensional gel electrophoresis and limited proteolysis. Suppression of PI1 synthesis in anterior fragments of normal embryos is ascribed to the activity of cytoplasmic ribonucleoprotein particles thought to act as anterior determinants. Images PMID:6935679

Third prize: the impact of fluid environment manipulation on shockwave lithotripsy artificial calculi fragmentation rates.

PubMed

Méndez-Probst, Carlos E; Fernadez, Alfonso; Erdeljan, Petar; Vanjecek, Maaike; Cadieux, Peter A; Razvi, Hassan

2011-03-01

Studies have suggested that shockwave lithotripsy (SWL) stone fragmentation rates can be affected by characteristics of the fluid media surrounding the stone, although evidence to implicate the impact of urine specific gravity (SG) is limited and inconclusive. Our aim is to further explore the impact fluid media and SGs have on stone fragmentation using a variable focus lithotripter. Artificial stones were presoaked for 24 hours in urine and then shocked in various fluid media including artificial urine (SG 1.010 control, 1.020, and 1.07), human pooled urine (HPU), degassed HPU, Pentastarch, 100% and 30% contrast, degassed 30% contrast, 100% ethanol, deionized water (dH(2)O), degassed dH(2)O, 5% glucose, Ringer lactate, 0.9% saline, glycerol, whole blood, and lubricating gel. After soaking, SWL using the Modulith SLX-F2 electromagnetic lithotripter was performed. Fragments were dried and sieved using a 4-mm diameter opening grid. Fragments >4 mm were weighed and fragmentation coefficients (FCs) calculated (pre-SWL weight - post-SWL weight)/(pre-SWL weight) × 100. Fifteen stones were shocked for each fluid group. Fluid type, viscosity, and degassing all significantly impacted stone fragmentation. While the solutions' SG, per se, did not appear to affect stone fragmentation, the use of degassed 30% contrast significantly improved stone destruction over the SG 1.010 artificial urine control (95.3% vs 71.4, P < 0.01). Furthermore, degassing improved comminution rates by increasing the number of completely fragmented stones (FC = 100%). Using degassed 30% contrast, 12/15 stones were completely fragmented, compared with only 2/15 in the control group (P = 0.007). Among the whole blood, glycerol, and lubricating gel groups, only 1/15, 0/15, and 1/15 stones reached 100% FC respectively in the narrow focus, possibly because of the detrimental impact of increased viscosity. Different fluid media can significantly affect FC in vitro. Among the various fluids tested, degassed 30% contrast significantly increased the FC and total number of completely fragmented stones.

The effects of habitat fragmentation on the social kin structure and mating system of the agile antechinus, Antechinus agilis.

PubMed

Banks, S C; Ward, S J; Lindenmayer, D B; Finlayson, G R; Lawson, S J; Taylor, A C

2005-05-01

Habitat fragmentation is one of the major contributors to the loss of biodiversity worldwide. However, relatively little is known about its more immediate impacts on within-patch population processes such as social structure and mating systems, whose alteration may play an important role in extinction risk. We investigated the impacts of habitat fragmentation due to the establishment of an exotic softwood plantation on the social kin structure and breeding system of the Australian marsupial carnivore, Antechinus agilis. Restricted dispersal by males in fragmented habitat resulted in elevated relatedness among potential mates in populations in fragments, potentially increasing the risk of inbreeding. Antechinus agilis nests communally in tree hollows; these nests are important points for social contact between males and females in the mating season. In response to elevated relatedness among potential mates in fragmented habitat, A. agilis significantly avoided sharing nests with opposite-sex relatives in large fragment sites (but not in small ones, possibly due to limited nest locations and small population sizes). Because opposite-sex individuals shared nests randomly with respect to relatedness in unfragmented habitat, we interpreted the phenomenon in fragmented habitat as a precursor to inbreeding avoidance via mate choice. Despite evidence that female A. agilis at high inbreeding risk selected relatively unrelated mates, there was no overall increased avoidance of related mates by females in fragmented habitats compared to unfragmented habitats. Simulations indicated that only dispersal, and not nonrandom mating, contributed to inbreeding avoidance in either habitat context. However, habitat fragmentation did influence the mating system in that the degree of multiple paternity was reduced due to the reduction in population sizes and population connectivity. This, in turn, reduced the number of males available to females in the breeding season. This suggests that in addition to the obvious impacts of reduced recruitment, patch recolonization and increased genetic drift, the isolation of populations in habitat patches may cause changes in breeding behaviour that contribute to the negative impacts of habitat fragmentation.

Volcanology: Petit spots go big

NASA Astrophysics Data System (ADS)

Snow, Jonathan E.

2016-12-01

Mantle enrichment processes were thought to be limited to parts of oceanic plates influenced by plumes and to continental interiors. Analyses of mantle fragments of the Pacific Plate suggest that such enrichment processes may operate everywhere.

Significance of the Fragmentation Region in Ultrarelativistic Heavy-Ion Collisions

NASA Astrophysics Data System (ADS)

Back, B. B.; Baker, M. D.; Barton, D. S.; Betts, R. R.; Ballintijn, M.; Bickley, A. A.; Bindel, R.; Budzanowski, A.; Busza, W.; Carroll, A.; Decowski, M. P.; García, E.; George, N.; Gulbrandsen, K.; Gushue, S.; Halliwell, C.; Hamblen, J.; Heintzelman, G. A.; Henderson, C.; Hofman, D. J.; Hollis, R. S.; Hołyński, R.; Holzman, B.; Iordanova, A.; Johnson, E.; Kane, J. L.; Katzy, J.; Khan, N.; Kucewicz, W.; Kulinich, P.; Kuo, C. M.; Lin, W. T.; Manly, S.; McLeod, D.; Michałowski, J.; Mignerey, A. C.; Nouicer, R.; Olszewski, A.; Pak, R.; Park, I. C.; Pernegger, H.; Reed, C.; Remsberg, L. P.; Reuter, M.; Roland, C.; Roland, G.; Rosenberg, L.; Sagerer, J.; Sarin, P.; Sawicki, P.; Skulski, W.; Steadman, S. G.; Steinberg, P.; Stephans, G. S.; Stodulski, M.; Sukhanov, A.; Tang, J.-L.; Teng, R.; Trzupek, A.; Vale, C.; van Nieuwenhuizen, G. J.; Verdier, R.; Wadsworth, B.; Wolfs, F. L.; Wosiek, B.; Woźniak, K.; Wuosmaa, A. H.; Wysłouch, B.

2003-08-01

We present measurements of the pseudorapidity distribution of primary charged particles produced in Au+Au collisions at three energies, (sNN)=19.6, 130, and 200GeV, for a range of collision centrali­ties. The distribution narrows for more central collisions and excess particles are produced at high pseudorapidity in peripheral collisions. For a given centrality, however, the distributions are found to scale with energy according to the “limiting fragmentation” hypothesis. The universal fragmentation region described by this scaling grows in pseudorapidity with increasing collision energy, extending well away from the beam rapidity and covering more than half of the pseudorapidity range over which particles are produced. This approach to a universal limiting curve appears to be a dominant feature of the pseudorapidity distribution and therefore of the total particle production in these collisions.

Demonstration of protein-fragment complementation assay using purified firefly luciferase fragments

PubMed Central

2013-01-01

Background Human interactome is predicted to contain 150,000 to 300,000 protein-protein interactions, (PPIs). Protein-fragment complementation assay (PCA) is one of the most widely used methods to detect PPI, as well as Förster resonance energy transfer (FRET). To date, successful applications of firefly luciferase (Fluc)-based PCA have been reported in vivo, in cultured cells and in cell-free lysate, owing to its high sensitivity, high signal-to-background (S/B) ratio, and reversible response. Here we show the assay also works with purified proteins with unexpectedly rapid kinetics. Results Split Fluc fragments both fused with a rapamycin-dependently interacting protein pair were made and expressed in E. coli system, and purified to homogeneity. When the proteins were used for PCA to detect rapamycin-dependent PPI, they enabled a rapid detection (~1 s) of PPI with high S/B ratio. When Fn7-8 domains (7 nm in length) that was shown to abrogate GFP mutant-based FRET was inserted between split Fluc and FKBP12 as a rigid linker, it still showed some response, suggesting less limitation in interacting partner’s size. Finally, the stability of the probe was investigated. Preincubation of the probes at 37 degreeC up to 1 h showed marked decrease of the luminescent signal to 1.5%, showing the limited stability of this system. Conclusion Fluc PCA using purified components will enable a rapid and handy detection of PPIs with high S/B ratio, avoiding the effects of concomitant components. Although the system might not be suitable for large-scale screening due to its limited stability, it can detect an interaction over larger distance than by FRET. This would be the first demonstration of Fluc PCA in vitro, which has a distinct advantage over other PPI assays. Our system enables detection of direct PPIs without risk of perturbation by PPI mediators in the complex cellular milieu. PMID:23536995

Effects of herbivory and patch size on tree seedling survivorship in a fog-dependent coastal rainforest in semiarid Chile.

PubMed

del-Val, Ek; Armesto, Juan J; Barbosa, Olga; Marquet, Pablo A

2007-09-01

The landscape (matrix) surrounding habitat fragments critically affects the biodiversity of those fragments due to biotic interchange and physical effects. However, to date, there have been only a limited number of studies on plant-animal interactions in fragmented landscapes, particularly on how tree seedling herbivory is affected by fragmentation. We have examined this question in a fog-dependent mosaic of rainforest fragments located on coastal mountaintops of semiarid Chile (30 degrees S), where the effects of the surrounding semiarid matrix and forest patch size (0.1-22 ha) on tree seedling survival were simultaneously addressed. The rainforest is strongly dominated by the endemic evergreen tree species Aextoxicon punctatum (Olivillo, approx. 80% of basal area). To assess the magnitudes and causes of Olivillo seedling mortality, we set up a field experiment where 512 tree seedlings of known age were transplanted into four forest fragments of different sizes in four 1.5 x 3-m plots per patch; one-half of each plot was fenced off with chicken wire to exclude small mammals. The plots were monitored for 22 months. Overall, 50% of the plants died during the experiment. The exclusion of small mammals from the plots increased seedling survival by 25%, with the effect being greater in smaller patches where matrix-dwelling herbivores are more abundant. This experiment highlights the important role of the surrounding matrix in affecting the persistence of trees in forest fragments. Because herbivores from the matrix cause greater tree seedling mortality in small patches, their effects must be taken into account in forest conservation-restoration plans.

A molecular fragment cheminformatics roadmap for mesoscopic simulation.

PubMed

Truszkowski, Andreas; Daniel, Mirco; Kuhn, Hubert; Neumann, Stefan; Steinbeck, Christoph; Zielesny, Achim; Epple, Matthias

2014-12-01

Mesoscopic simulation studies the structure, dynamics and properties of large molecular ensembles with millions of atoms: Its basic interacting units (beads) are no longer the nuclei and electrons of quantum chemical ab-initio calculations or the atom types of molecular mechanics but molecular fragments, molecules or even larger molecular entities. For its simulation setup and output a mesoscopic simulation kernel software uses abstract matrix (array) representations for bead topology and connectivity. Therefore a pure kernel-based mesoscopic simulation task is a tedious, time-consuming and error-prone venture that limits its practical use and application. A consequent cheminformatics approach tackles these problems and provides solutions for a considerably enhanced accessibility. This study aims at outlining a complete cheminformatics roadmap that frames a mesoscopic Molecular Fragment Dynamics (MFD) simulation kernel to allow its efficient use and practical application. The molecular fragment cheminformatics roadmap consists of four consecutive building blocks: An adequate fragment structure representation (1), defined operations on these fragment structures (2), the description of compartments with defined compositions and structural alignments (3), and the graphical setup and analysis of a whole simulation box (4). The basis of the cheminformatics approach (i.e. building block 1) is a SMILES-like line notation (denoted f SMILES) with connected molecular fragments to represent a molecular structure. The f SMILES notation and the following concepts and methods for building blocks 2-4 are outlined with examples and practical usage scenarios. It is shown that the requirements of the roadmap may be partly covered by already existing open-source cheminformatics software. Mesoscopic simulation techniques like MFD may be considerably alleviated and broadened for practical use with a consequent cheminformatics layer that successfully tackles its setup subtleties and conceptual usage hurdles. Molecular Fragment Cheminformatics may be regarded as a crucial accelerator to propagate MFD and similar mesoscopic simulation techniques in the molecular sciences. Graphical abstractA molecular fragment cheminformatics roadmap for mesoscopic simulation.

Integrative Review of the Literature on Adults with Limited Education and Skills and the Implications for Human Resource Development

ERIC Educational Resources Information Center

Hamilton, David W.; Torraco, Richard J.

2013-01-01

Adults with limited education and skills--those who lack the education and skills needed for full participation in U.S. culture and economy--are increasing in numbers. However, the knowledge base addressing this population and their educational needs is fragmented across the literature of several disciplines. A comprehensive review and critique of…

A species on a tightrope: Establishment limitations of an endangered lichen in a fragmented Mediterranean landscape.

PubMed

Cardós, Juan Luis H; Aragón, Gregorio; Martínez, Isabel

2017-04-01

Habitat loss and forest fragmentation affect the dispersal and establishment of species. Furthermore, populations growing far from the species' optimal climate might be less viable because good-quality habitat can be scarce and easily altered by smaller changes. The lichen Pectenia plumbea has oceanic climatic requirements, so in the Mediterranean region it needs the humidity provided by well-preserved forests to thrive, but most of this habitat has disappeared and the remnants are fragmented. In central Spain, this species occupies only a small proportion of the existing forests, so we aimed to determine whether this scattered distribution is due to limitations on dispersal or establishment. We selected a Mediterranean fragmented forest surface in central Spain and extracted environmental variables from 371 plots. We modeled the presence and abundance of P. plumbea and developed species distribution models (SDMs) to detect all the suitable habitats inside the Cabañeros National Park area. Pectenia plumbea was present in most of the habitats predicted as good-quality and was generally absent from the poor-quality zones (85.9% overall success). The abundance correlated fairly well with that predicted by the SDM (67%). Both models show that P. plumbea is linked to high temperature and precipitation. Good-quality habitat requirements for P. plumbea that are similar to oceanic conditions are found only in specific forested, stony slopes derived from historical land management. This habitat is scarce, but P. plumbea has successfully tracked all of these scattered areas via its high dispersal capacity. © 2017 Botanical Society of America.

Threatened and Endangered Subspecies with Vulnerable Ecological Traits Also Have High Susceptibility to Sea Level Rise and Habitat Fragmentation

PubMed Central

Benscoter, Allison M.; Reece, Joshua S.; Noss, Reed F.; Brandt, Laura A.; Mazzotti, Frank J.; Romañach, Stephanie S.; Watling, James I.

2013-01-01

The presence of multiple interacting threats to biodiversity and the increasing rate of species extinction make it critical to prioritize management efforts on species and communities that maximize conservation success. We implemented a multi-step approach that coupled vulnerability assessments evaluating threats to Florida taxa such as climate change, sea-level rise, and habitat fragmentation with in-depth literature surveys of taxon-specific ecological traits. The vulnerability, adaptive capacity, and ecological traits of 12 threatened and endangered subspecies were compared to non-listed subspecies of the same parent species. Overall, the threatened and endangered subspecies showed high vulnerability and low adaptive capacity, in particular to sea level rise and habitat fragmentation. They also exhibited larger home ranges and greater dispersal limitation compared to non-endangered subspecies, which may inhibit their ability to track changing climate in fragmented landscapes. There was evidence for lower reproductive capacity in some of the threatened or endangered taxa, but not for most. Taxa located in the Florida Keys or in other low coastal areas were most vulnerable to sea level rise, and also showed low levels of adaptive capacity, indicating they may have a lower probability of conservation success. Our analysis of at-risk subspecies and closely related non-endangered subspecies demonstrates that ecological traits help to explain observed differences in vulnerability and adaptive capacity. This study points to the importance of assessing the relative contributions of multiple threats and evaluating conservation value at the species (or subspecies) level when resources are limited and several factors affect conservation success. PMID:23940614

The Effects of Trivalent Lanthanide Cationization on the Electron Transfer Dissociation of Acidic Fibrinopeptide B and its Analogs

NASA Astrophysics Data System (ADS)

Commodore, Juliette J.; Cassady, Carolyn J.

2016-09-01

Electrospray ionization (ESI) on mixtures of acidic fibrinopeptide B and two peptide analogs with trivalent lanthanide salts generates [M + Met + H]4+, [M + Met]3+, and [M + Met -H]2+, where M = peptide and Met = metal (except radioactive promethium). These ions undergo extensive and highly efficient electron transfer dissociation (ETD) to form metallated and non-metallated c- and z-ions. All metal adducted product ions contain at least two acidic sites, which suggest attachment of the lanthanide cation at the side chains of one or more acidic residues. The three peptides undergo similar fragmentation. ETD on [M + Met + H]4+ leads to cleavage at every residue; the presence of both a metal ion and an extra proton is very effective in promoting sequence-informative fragmentation. Backbone dissociation of [M + Met]3+ is also extensive, although cleavage does not always occur between adjacent glutamic acid residues. For [M + Met - H ]2+, a more limited range of product ions form. All lanthanide metal peptide complexes display similar fragmentation except for europium (Eu). ETD on [M + Eu - H]2+ and [M + Eu]3+ yields a limited amount of peptide backbone cleavage; however, [M + Eu + H]4+ dissociates extensively with cleavage at every residue. With the exception of the results for Eu(III), metallated peptide ion formation by ESI, ETD fragmentation efficiencies, and product ion formation are unaffected by the identity of the lanthanide cation. Adduction with trivalent lanthanide metal ions is a promising tool for sequence analysis of acidic peptides by ETD.

Minimizing back exchange in the hydrogen exchange-mass spectrometry experiment.

PubMed

Walters, Benjamin T; Ricciuti, Alec; Mayne, Leland; Englander, S Walter

2012-12-01

The addition of mass spectrometry (MS) analysis to the hydrogen exchange (HX) proteolytic fragmentation experiment extends powerful HX methodology to the study of large biologically important proteins. A persistent problem is the degradation of HX information due to back exchange of deuterium label during the fragmentation-separation process needed to prepare samples for MS measurement. This paper reports a systematic analysis of the factors that influence back exchange (solution pH, ionic strength, desolvation temperature, LC column interaction, flow rates, system volume). The many peptides exhibit a range of back exchange due to intrinsic amino acid HX rate differences. Accordingly, large back exchange leads to large variability in D-recovery from one residue to another as well as one peptide to another that cannot be corrected for by reference to any single peptide-level measurement. The usual effort to limit back exchange by limiting LC time provides little gain. Shortening the LC elution gradient by 3-fold only reduced back exchange by ~2%, while sacrificing S/N and peptide count. An unexpected dependence of back exchange on ionic strength as well as pH suggests a strategy in which solution conditions are changed during sample preparation. Higher salt should be used in the first stage of sample preparation (proteolysis and trapping) and lower salt (<20 mM) and pH in the second stage before electrospray injection. Adjustment of these and other factors together with recent advances in peptide fragment detection yields hundreds of peptide fragments with D-label recovery of 90% ± 5%.

Threatened and endangered subspecies with vulnerable ecological traits also have high susceptibility to sea level rise and habitat fragmentation.

PubMed

Benscoter, Allison M; Reece, Joshua S; Noss, Reed F; Brandt, Laura A; Mazzotti, Frank J; Romañach, Stephanie S; Watling, James I

2013-01-01

The presence of multiple interacting threats to biodiversity and the increasing rate of species extinction make it critical to prioritize management efforts on species and communities that maximize conservation success. We implemented a multi-step approach that coupled vulnerability assessments evaluating threats to Florida taxa such as climate change, sea-level rise, and habitat fragmentation with in-depth literature surveys of taxon-specific ecological traits. The vulnerability, adaptive capacity, and ecological traits of 12 threatened and endangered subspecies were compared to non-listed subspecies of the same parent species. Overall, the threatened and endangered subspecies showed high vulnerability and low adaptive capacity, in particular to sea level rise and habitat fragmentation. They also exhibited larger home ranges and greater dispersal limitation compared to non-endangered subspecies, which may inhibit their ability to track changing climate in fragmented landscapes. There was evidence for lower reproductive capacity in some of the threatened or endangered taxa, but not for most. Taxa located in the Florida Keys or in other low coastal areas were most vulnerable to sea level rise, and also showed low levels of adaptive capacity, indicating they may have a lower probability of conservation success. Our analysis of at-risk subspecies and closely related non-endangered subspecies demonstrates that ecological traits help to explain observed differences in vulnerability and adaptive capacity. This study points to the importance of assessing the relative contributions of multiple threats and evaluating conservation value at the species (or subspecies) level when resources are limited and several factors affect conservation success.

Minimizing Back Exchange in the Hydrogen Exchange-Mass Spectrometry Experiment

NASA Astrophysics Data System (ADS)

Walters, Benjamin T.; Ricciuti, Alec; Mayne, Leland; Englander, S. Walter

2012-12-01

The addition of mass spectrometry (MS) analysis to the hydrogen exchange (HX) proteolytic fragmentation experiment extends powerful HX methodology to the study of large biologically important proteins. A persistent problem is the degradation of HX information due to back exchange of deuterium label during the fragmentation-separation process needed to prepare samples for MS measurement. This paper reports a systematic analysis of the factors that influence back exchange (solution pH, ionic strength, desolvation temperature, LC column interaction, flow rates, system volume). The many peptides exhibit a range of back exchange due to intrinsic amino acid HX rate differences. Accordingly, large back exchange leads to large variability in D-recovery from one residue to another as well as one peptide to another that cannot be corrected for by reference to any single peptide-level measurement. The usual effort to limit back exchange by limiting LC time provides little gain. Shortening the LC elution gradient by 3-fold only reduced back exchange by ~2 %, while sacrificing S/N and peptide count. An unexpected dependence of back exchange on ionic strength as well as pH suggests a strategy in which solution conditions are changed during sample preparation. Higher salt should be used in the first stage of sample preparation (proteolysis and trapping) and lower salt (<20 mM) and pH in the second stage before electrospray injection. Adjustment of these and other factors together with recent advances in peptide fragment detection yields hundreds of peptide fragments with D-label recovery of 90 % ± 5 %.

Threatened and endangered subspecies with vulnerable ecological traits Also have high susceptibility to sea level rise and habitat fragmentation

USGS Publications Warehouse

Benscoter, Allison M.; Reece, Joshua S.; Noss, Reed F.; Brandt, Laura B.; Mazzotti, Frank J.; Romañach, Stephanie S.; Watling, James I.

2013-01-01

The presence of multiple interacting threats to biodiversity and the increasing rate of species extinction make it critical to prioritize management efforts on species and communities that maximize conservation success. We implemented a multi-step approach that coupled vulnerability assessments evaluating threats to Florida taxa such as climate change, sea-level rise, and habitat fragmentation with in-depth literature surveys of taxon-specific ecological traits. The vulnerability, adaptive capacity, and ecological traits of 12 threatened and endangered subspecies were compared to non-listed subspecies of the same parent species. Overall, the threatened and endangered subspecies showed high vulnerability and low adaptive capacity, in particular to sea level rise and habitat fragmentation. They also exhibited larger home ranges and greater dispersal limitation compared to non-endangered subspecies, which may inhibit their ability to track changing climate in fragmented landscapes. There was evidence for lower reproductive capacity in some of the threatened or endangered taxa, but not for most. Taxa located in the Florida Keys or in other low coastal areas were most vulnerable to sea level rise, and also showed low levels of adaptive capacity, indicating they may have a lower probability of conservation success. Our analysis of at-risk subspecies and closely related non-endangered subspecies demonstrates that ecological traits help to explain observed differences in vulnerability and adaptive capacity. This study points to the importance of assessing the relative contributions of multiple threats and evaluating conservation value at the species (or subspecies) level when resources are limited and several factors affect conservation success.

How do protected landscapes associated with high biodiversity and population levels change?

PubMed Central

Echeverria, Cristian

2017-01-01

Most protected areas (PA) try to limit logging of forests by means of restrictions on access and use, especially in areas where local communities coexist with the forests and depend on resources derived from PAs. In such contexts, achieving full or effective protection of the forests is almost impossible. This fact has led to researching beyond PAs boundaries in order to examine large surrounding landscapes with multiple forms of properties and restriction on forests use. The present study assessed the change in forest cover and fragmentation between 1990 and 2014, in addition to the drivers that explain such changes in a landscape with the presence of PAs and high-density population belonging to the Chocó-Darien biodiversity hotspot. Results indicated differences in the extent and spatial patterns of change in forest cover of PAs and their surrounding landscapes. Two PAs exhibited a tendency to increase fragmentation and lose their forests in comparison with the stable protection of the forests in other PAs during this period. However, the greatest change in forest cover and fragmentation was observed in the surrounding landscapes, where the best connection to markets and transport networks were the dominating deforestation drivers. Our findings corroborated that the PAs were a shield against the deforestation of the tropical Andean forest, especially in landscapes with high-density population. However, the fragmentation of the forest cannot be avoided around the PAs limits. It is expected that, if this tendency continues in the future, the biodiversity in the Chocó-Darien hotspot will be seriously affected. PMID:28672002

DOE Office of Scientific and Technical Information (OSTI.GOV)

Scolozzi, Rocco, E-mail: rocco.scolozzi@fmach.it; Geneletti, Davide, E-mail: geneletti@ing.unitn.it

Habitat loss and fragmentation are often concurrent to land conversion and urbanization. Simple application of GIS-based landscape pattern indicators may be not sufficient to support meaningful biodiversity impact assessment. A review of the literature reveals that habitat definition and habitat fragmentation are frequently inadequately considered in environmental assessment, notwithstanding the increasing number of tools and approaches reported in the landscape ecology literature. This paper presents an approach for assessing impacts on habitats on a local scale, where availability of species data is often limited, developed for an alpine valley in northern Italy. The perspective of the methodology is multiple scalemore » and species-oriented, and provides both qualitative and quantitative definitions of impact significance. A qualitative decision model is used to assess ecological values in order to support land-use decisions at the local level. Building on recent studies in the same region, the methodology integrates various approaches, such as landscape graphs, object-oriented rule-based habitat assessment and expert knowledge. The results provide insights into future habitat loss and fragmentation caused by land-use changes, and aim at supporting decision-making in planning and suggesting possible ecological compensation. - Highlights: Black-Right-Pointing-Pointer Many environmental assessments inadequately consider habitat loss and fragmentation. Black-Right-Pointing-Pointer Species-perspective for defining habitat quality and connectivity is claimed. Black-Right-Pointing-Pointer Species-based tools are difficult to be applied with limited availability of data. Black-Right-Pointing-Pointer We propose a species-oriented and multiple scale-based qualitative approach. Black-Right-Pointing-Pointer Advantages include being species-oriented and providing value-based information.« less

Telomere Restriction Fragment (TRF) Analysis.

PubMed

Mender, Ilgen; Shay, Jerry W

2015-11-20

While telomerase is expressed in ~90% of primary human tumors, most somatic tissue cells except transiently proliferating stem-like cells do not have detectable telomerase activity (Shay and Wright, 1996; Shay and Wright, 2001). Telomeres progressively shorten with each cell division in normal cells, including proliferating stem-like cells, due to the end replication (lagging strand synthesis) problem and other causes such as oxidative damage, therefore all somatic cells have limited cell proliferation capacity (Hayflick limit) (Hayflick and Moorhead, 1961; Olovnikov, 1973). The progressive telomere shortening eventually leads to growth arrest in normal cells, which is known as replicative senescence (Shay et al. , 1991). Once telomerase is activated in cancer cells, telomere length is stabilized by the addition of TTAGGG repeats to the end of chromosomes, thus enabling the limitless continuation of cell division (Shay and Wright, 1996; Shay and Wright, 2001). Therefore, the link between aging and cancer can be partially explained by telomere biology. There are many rapid and convenient methods to study telomere biology such as Telomere Restriction Fragment (TRF), Telomere Repeat Amplification Protocol (TRAP) (Mender and Shay, 2015b) and Telomere dysfunction Induced Foci (TIF) analysis (Mender and Shay, 2015a). In this protocol paper we describe Telomere Restriction Fragment (TRF) analysis to determine average telomeric length of cells. Telomeric length can be indirectly measured by a technique called Telomere Restriction Fragment analysis (TRF). This technique is a modified Southern blot, which measures the heterogeneous range of telomere lengths in a cell population using the length distribution of the terminal restriction fragments (Harley et al. , 1990; Ouellette et al. , 2000). This method can be used in eukaryotic cells. The description below focuses on the measurement of human cancer cells telomere length. The principle of this method relies on the lack of restriction enzyme recognition sites within TTAGGG tandem telomeric repeats, therefore digestion of genomic DNA, not telomeric DNA, with a combination of 6 base restriction endonucleases reduces genomic DNA size to less than 800 bp.

Anticipated effects of development on habitat fragmentation and movement of mammals into and out of the Schoodic District, Acadia National Park, Maine

USGS Publications Warehouse

Rohweder, Jason J.; De Jager, Nathan R.; Guntenspergen, Glenn R.

2012-01-01

Most national parks interact with adjacent lands because their boundaries fail to encompass all regional habitats, species pools, and migration routes. Activities planned for adjacent lands can have adverse effects on park resources and visitor experiences. For example, fragmentation of adjacent habitat into smaller and more isolated remnants may influence the suitability of park habitat for a wide range of species and limit animal dispersal pathways, which may influence visitor experiences and park resources as well as the energy balance and population dynamics of the animals themselves. In this study, we examined habitat fragmentation consequences owing to a planned 1,295 hectare development by Winter Harbor Holding Company (WHHC) adjacent to the Schoodic District of Acadia National Park (ANP), Maine. Specifically, we examined the effects of development on (a) core natural habitat area (a cross-habitat indicator of fragmentation), (b) the suitability of habitat for bobcat, fisher, mink, and moose, and (c) the movement of these four species between ANP and other nearby protected areas (species specific indicators of fragmentation). Our intention was to assist ANP staff in forecasting both the general and specific effects of development on natural habitat area, habitat suitability, and animal movement, which would allow them to develop suitable management alternatives.

Fusion-fission and quasifission of superheavy systems with Z =110 -116 formed in 48Ca-induced reactions

NASA Astrophysics Data System (ADS)

Kozulin, E. M.; Knyazheva, G. N.; Itkis, I. M.; Itkis, M. G.; Bogachev, A. A.; Chernysheva, E. V.; Krupa, L.; Hanappe, F.; Dorvaux, O.; Stuttgé, L.; Trzaska, W. H.; Schmitt, C.; Chubarian, G.

2014-11-01

Background: In heavy-ion-induced reactions the mechanism leading to the formation of the compound nucleus and the role of quasifission is still not clear. Purpose: Investigation of the quasifission process of superheavy composite systems with Z =110 -116 and comparison with properties of fusion-fission and quasifission of lighter composite systems. Method: Mass and energy distributions of fissionlike fragments formed in the reactions 48Ca+232Th, 238U , 244Pu , and 248Cm at energies near the Coulomb barrier have been measured using the double-arm time-of-flight spectrometer CORSET at the U-400 cyclotron of the FLNR JINR. Results: The most probable fragment masses as well as total kinetic energies and their dispersions in dependence on the interaction energies and ion-target combinations have been studied for asymmetric and symmetric fragments formed in the reactions. The capture cross sections were obtained for the reactions 48Ca+244Pu and 248Cm . The lower limits for fission barriers of 283 -286Cn , 289 -292Fl , and 293 -296Lv compound nuclei were estimated. Conclusions: Analysis of the properties of symmetric fragments has shown that a significant part of these fragments may be attributed to fusion-fission process for the reactions 48Ca +238U , 244Pu , and 248Cm .

Experimental and data analysis techniques for deducing collision-induced forces from photographic histories of engine rotor fragment impact/interaction with a containment ring

NASA Technical Reports Server (NTRS)

Yeghiayan, R. P.; Leech, J. W.; Witmer, E. A.

1973-01-01

An analysis method termed TEJ-JET is described whereby measured transient elastic and inelastic deformations of an engine-rotor fragment-impacted structural ring are analyzed to deduce the transient external forces experienced by that ring as a result of fragment impact and interaction with the ring. Although the theoretical feasibility of the TEJ-JET concept was established, its practical feasibility when utilizing experimental measurements of limited precision and accuracy remains to be established. The experimental equipment and the techniques (high-speed motion photography) employed to measure the transient deformations of fragment-impacted rings are described. Sources of error and data uncertainties are identified. Techniques employed to reduce data reading uncertainties and to correct the data for optical-distortion effects are discussed. These procedures, including spatial smoothing of the deformed ring shape by Fourier series and timewise smoothing by Gram polynomials, are applied illustratively to recent measurements involving the impact of a single T58 turbine rotor blade against an aluminum containment ring. Plausible predictions of the fragment-ring impact/interaction forces are obtained by one branch of this TEJ-JET method; however, a second branch of this method, which provides an independent estimate of these forces, remains to be evaluated.

Understanding the Physical Structure of the Comet Shoemaker-Levy 9 Fragments

NASA Astrophysics Data System (ADS)

Rettig, Terrence

2000-07-01

Images of the fragmented comet Shoemaker-Levy 9 {SL9} as it approached Jupiter in 1994 provided a unique opportunity to {1} probe the comae, {2} understand the structure of the 20 cometary objects, and {3} provide limits on the Jovian impact parameters. The primary cometary questions were: how were the fragments formed and what was their central structure? There still remains a diversity of opinion regarding the structure of the 21 comet-like fragments as well as the specifics of the disruption event itself. We have shown from Monte Carlo modeling of surface brightness profiles that SL9 fragments had unusual dust size distributions and outflow velocities. Further work of a preliminary nature showed some of the central reflecting area excesses derived from surface brightness profile fitting {w/psf} appeared distributed rather than centrally concentrated as would be expected for comet- like objects, some central excesses were negative and also, the excesses could vary with time. With an improved coma subtraction technique we propose to model each coma surface brightness profile, extract central reflecting areas or central brightness excesses for the non-star-contaminated WFPC-2 SL9, to determine the behavior and characteristics of the central excesses as the fragments approached Jupiter. A second phase of the proposal will be to use numerical techniques {in conjunction with D. Richardson} to investigate the various fragment models. This is a difficult modeling process that will allow us to model the structure and physical characteristics of the fragments and thus constrain parameters for the Jovian impact events. The results will be used to constrain the structure of the central fragment cores of SL9 and how the observed dust comae were produced. The results will provide evidence to discriminate between the parent nucleus models {i.e., were the fragments solid objects or swarms of particles?} and provide better constraints on the atmospheric impact models. The physical characteristics of cometary nuclei are not well understood and the SL9 data provides an important opportunity to constrain these parameters.

Fragment Based Optimization of Metabotropic Glutamate Receptor 2 (mGluR2) Positive Allosteric Modulators in the Absence of Structural Information.

PubMed

Szabó, György; Túrós, György I; Kolok, Sándor; Vastag, Mónika; Sánta, Zsuzsanna; Dékány, Miklós; Lévay, György I; Greiner, István; Natsumi, Minami; Tatsuya, Watanabe; Keserű, György M

2018-03-14

Metabotropic glutamate receptor 2 (mGluR2) positive allosteric modulators (PAMs) have been implicated as potential pharmacotherapy for psychiatric conditions. Screening our corporate compound deck, we identified a benzotriazole fragment (4) that was rapidly optimized to a potent and metabolically stable early lead (16). The highly lipophilic character of 16, together with its limited solubility, permeability, and high protein binding, however, did not allow reaching of the proof of concept in vivo. Since further attempts on the optimization of druglike properties were unsuccessful, the original hit 4 has been revisited and was optimized following the principles of fragment based drug discovery (FBDD). Lacking structural information on the receptor-ligand complex, we implemented a group efficiency (GE) based strategy and identified a new fragment like lead (60) with more balanced profile. Significant improvement achieved on the druglike properties nominated the compound for in vivo proof of concept studies that revealed the chemotype being a promising PAM lead targeting mGluR2 receptors.

Research on memory management in embedded systems

NASA Astrophysics Data System (ADS)

Huang, Xian-ying; Yang, Wu

2005-12-01

Memory is a scarce resource in embedded system due to cost and size. Thus, applications in embedded systems cannot use memory randomly, such as in desktop applications. However, data and code must be stored into memory for running. The purpose of this paper is to save memory in developing embedded applications and guarantee running under limited memory conditions. Embedded systems often have small memory and are required to run a long time. Thus, a purpose of this study is to construct an allocator that can allocate memory effectively and bear a long-time running situation, reduce memory fragmentation and memory exhaustion. Memory fragmentation and exhaustion are related to the algorithm memory allocated. Static memory allocation cannot produce fragmentation. In this paper it is attempted to find an effective allocation algorithm dynamically, which can reduce memory fragmentation. Data is the critical part that ensures an application can run regularly, which takes up a large amount of memory. The amount of data that can be stored in the same size of memory is relevant with the selected data structure. Skills for designing application data in mobile phone are explained and discussed also.

Targeting α-synuclein oligomers by protein-fragment complementation for drug discovery in synucleinopathies.

PubMed

Moussaud, Simon; Malany, Siobhan; Mehta, Alka; Vasile, Stefan; Smith, Layton H; McLean, Pamela J

2015-05-01

Reducing the burden of α-synuclein oligomeric species represents a promising approach for disease-modifying therapies against synucleinopathies such as Parkinson's disease and dementia with Lewy bodies. However, the lack of efficient drug discovery strategies that specifically target α-synuclein oligomers has been a limitation to drug discovery programs. Here we describe an innovative strategy that harnesses the power of bimolecular protein-fragment complementation to monitor synuclein-synuclein interactions. We have developed two robust models to monitor α-synuclein oligomerization by generating novel stable cell lines expressing α-synuclein fusion proteins for either fluorescent or bioluminescent protein-fragment complementation under the tetracycline-controlled transcriptional activation system. A pilot screen was performed resulting in the identification of two potential hits, a p38 MAPK inhibitor and a casein kinase 2 inhibitor, thereby demonstrating the suitability of our protein-fragment complementation assay for the measurement of α-synuclein oligomerization in living cells at high throughput. The application of the strategy described herein to monitor α-synuclein oligomer formation in living cells with high throughput will facilitate drug discovery efforts for disease-modifying therapies against synucleinopathies and other proteinopathies.

NMR characterization of weak interactions between RhoGDI2 and fragment screening hits.

PubMed

Liu, Jiuyang; Gao, Jia; Li, Fudong; Ma, Rongsheng; Wei, Qingtao; Wang, Aidong; Wu, Jihui; Ruan, Ke

2017-01-01

The delineation of intrinsically weak interactions between novel targets and fragment screening hits has long limited the pace of hit-to-lead evolution. Rho guanine-nucleotide dissociation inhibitor 2 (RhoGDI2) is a novel target that lacks any chemical probes for the treatment of tumor metastasis. Protein-observed and ligand-observed NMR spectroscopy was used to characterize the weak interactions between RhoGDI2 and fragment screening hits. We identified three hits of RhoGDI2 using streamlined NMR fragment-based screening. The binding site residues were assigned using non-uniformly sampled C α - and H α -based three dimensional NMR spectra. The molecular docking to the proposed geranylgeranyl binding pocket of RhoGDI2 was guided by NMR restraints of chemical shift perturbations and ligand-observed transferred paramagnetic relaxation enhancement. We further validated the weak RhoGDI2-hit interactions using mutagenesis and structure-affinity analysis. Weak interactions between RhoGDI2 and fragment screening hits were delineated using an integrated NMR approach. Binders to RhoGDI2 as a potential anti-cancer target have been first reported, and their weak interactions were depicted using NMR spectroscopy. Our work highlights the powerfulness and the versatility of the integrative NMR techniques to provide valuable structural insight into the intrinsically weak interactions between RhoGDI2 and the fragment screening hits, which could hardly be conceived using other biochemical techniques. Copyright © 2016 Elsevier B.V. All rights reserved.

Use of CID/ETD Mass Spectrometry to Analyze Glycopeptides

PubMed Central

Mechref, Yehia

2013-01-01

Collision-induced dissociation (CID) tandem mass spectrometry (MS) does not allow the characterization of glycopeptides because of the fragmentation of their glycan structures and limited fragmentation of peptide backbones. Electron-transfer dissociation (ETD) tandem MS, on the other hand, offers an alternative approach allowing the fragmentation of only peptide backbones of glycopeptides. Characterization of glycopeptides using both CID and ETD is summarized in this unit. While CID provide information related to the composition of glycan moiety attached to a peptide backbone, ETD permits de novo sequencing of peptides, since it prompts only peptide backbone fragmentation while keeping posttranslational modifications intact. Radical anions transfer of electrons to peptide backbone which induces cleavage of the N-Cα bond is observed in ETD. The glycan moiety is retained on the peptide backbone, largely unaffected by the ETD process. Accordingly, ETD allows not only the identification of the amino acid sequence of a glycopeptide, but also the unambiguous assignment of its glycosylation site. When data acquired from both fragmentation techniques are combined, it is possible to characterize comprehensively the entire glycopeptide. This is achieved using an instrument capable of alternating between CID and ETD experiments during an LC-MS/MS analysis. This unit discusses the different fragmentation of glycopeptides observed in CID and ETD. Tables of residue masses associated with oxonium ions observed in CID are provided to help in the interpretation of CID mass spectra. The utility of both CID and ETD for better characterization of glycopeptides are demonstrated for a model glycoprotein. PMID:22470127

The influence of habitat fragmentation on helminth communities in rodent populations from a Brazilian Mountain Atlantic Forest.

PubMed

Cardoso, T S; Simões, R O; Luque, J L F; Maldonado, A; Gentile, R

2016-07-01

The influence of habitat structure on helminth communities of three sigomdontinae rodent species (Akodon cursor, A. montensis and Oligoryzomys nigripes) was investigated in forest fragments within an agricultural landscape in south-eastern Brazil. This is a pionner study correlating the occurrence of helminth species of rodent hosts with microhabitat characteristics. Rodents were collected from 12 fragments and in a continuous conserved area. Up to 13 nematode, three cestode and two trematode species were identified, and habitat fragmentation was found to have more influence on the helminth composition of O. nigripes compared to the other two rodent species. Fragmentation appeared to limit the development of some helminths' life cycles, e.g. with some species such as Trichofreitasia lenti, Protospirura numidica, Cysticercus fasciolaris and Avellaria sp., occurring mostly in areas with less anthropic impact. However, fragmentation did not seem to affect the life cycles of other dominant helminths, such as the trematode Canaania obesa, the nematodes Stilestrongylus lanfrediae, S. eta and S. aculeata, and the cestode Rodentolepis akodontis. The helminth community structure followed a nested pattern of distribution in A. montensis and O. nigripes. Stilestrongylus lanfrediae seemed to be more associated with dense understorey, C. obesa with open canopy and dense understorey, and Guerrerostrongylus zetta with organic matter on the ground. Their presence in each area may be explained by aspects of their life cycles that take place in the external environment outside the host.

A SQUID-based metal detector—comparison to coil and x-ray systems

NASA Astrophysics Data System (ADS)

Bick, M.; Sullivan, P.; Tilbrook, D. L.; Du, J.; Gnanarajan, S.; Leslie, K. E.; Foley, C. P.

2005-03-01

The presence of foreign metal bodies and fragments in foodstuff and pharmaceutical products is of major concern to producers. Further, hidden metal objects can pose threats to security. In particular, stainless steel is difficult to detect by conventional coil metal detectors due to its low conductivity. We have employed an HTS SQUID magnetometer for the detection of stainless steel particles which is based on the measurement of the remanent magnetization of the particle. Our aim was to determine the detection limits of HTS SQUID-based remote magnetometry, especially for food inspection purposes, and to make a comparison of this technique to commonly used eddy current coil and x-ray inspection systems. We show that the SQUID system's sensitivity to stainless steel fragments is significantly higher than that of coil systems if the samples are magnetized in a 100 mT magnetic field prior to detection. Further, it has a higher sensitivity than x-ray systems, depending on the density distribution of the product under inspection. A 0.6 mg piece of grade-316 stainless steel (a fragment of a hypodermic needle 0.5 mm long and 0.65 mm diameter) represents the detection limit of our system with a 150 × 150 mm2 inspection orifice.

Three new sensitive and specific heat-shock protein 70 PCRs for global Leishmania species identification.

PubMed

Montalvo, A M; Fraga, J; Maes, I; Dujardin, J-C; Van der Auwera, G

2012-07-01

The heat-shock protein 70 gene (hsp70) has been exploited for Leishmania species identification in the New and Old World, using PCR followed by restriction fragment length polymorphism (RFLP) analysis. Current PCR presents limitations in terms of sensitivity, which hampers its use for analyzing clinical and biological samples, and specificity, which makes it inappropriate to discriminate between Leishmania and other trypanosomatids. The aim of the study was to improve the sensitivity and specificity of a previously reported hsp70 PCR using alternative PCR primers and RFLPs. Following in silico analysis of available sequences, three new PCR primer sets and restriction digest schemes were tested on a globally representative panel of 114 Leishmania strains, various other infectious agents, and clinical samples. The largest new PCR fragment retained the discriminatory power from RFLP, while two smaller fragments discriminated less species. The detection limit of the new PCRs was between 0.05 and 0.5 parasite genomes, they amplified clinical samples more efficiently, and were Leishmania specific. We succeeded in significantly improving the specificity and sensitivity of the PCRs for hsp70 Leishmania species typing. The improved PCR-RFLP assays can impact diagnosis, treatment, and epidemiological studies of leishmaniasis in any setting worldwide.

Time-Lag in Responses of Birds to Atlantic Forest Fragmentation: Restoration Opportunity and Urgency.

PubMed

Uezu, Alexandre; Metzger, Jean Paul

2016-01-01

There are few opportunities to evaluate the relative importance of landscape structure and dynamics upon biodiversity, especially in highly fragmented tropical landscapes. Conservation strategies and species risk evaluations often rely exclusively on current aspects of landscape structure, although such limited assumptions are known to be misleading when time-lag responses occur. By relating bird functional-group richness to forest patch size and isolation in ten-year intervals (1956, 1965, 1978, 1984, 1993 and 2003), we revealed that birds with different sensitivity to fragmentation display contrasting responses to landscape dynamics in the Brazilian Atlantic Forest. For non-sensitive groups, there was no time-lag in response: the recent degree of isolation best explains their variation in richness, which likely relates to these species' flexibility to adapt to changes in landscape structure. However, for sensitive bird groups, the 1978 patch area was the best explanatory variable, providing evidence for a 25-year time-lag in response to habitat reduction. Time-lag was more likely in landscapes that encompass large patches, which can support temporarily the presence of some sensitive species, even when habitat cover is relatively low. These landscapes potentially support the most threatened populations and should be priorities for restoration efforts to avoid further species loss. Although time-lags provide an opportunity to counteract the negative consequences of fragmentation, it also reinforces the urgency of restoration actions. Fragmented landscapes will be depleted of biodiversity if landscape structure is only maintained, and not improved. The urgency of restoration action may be even higher in landscapes where habitat loss and fragmentation history is older and where no large fragment remained to act temporarily as a refuge.

Negative Impacts of Human Land Use on Dung Beetle Functional Diversity

PubMed Central

Barragán, Felipe; Moreno, Claudia E.; Escobar, Federico; Halffter, Gonzalo; Navarrete, Dario

2011-01-01

The loss of biodiversity caused by human activity is assumed to alter ecosystem functioning. However our understanding of the magnitude of the effect of these changes on functional diversity and their impact on the dynamics of ecological processes is still limited. We analyzed the functional diversity of copro-necrophagous beetles under different conditions of land use in three Mexican biosphere reserves. In Montes Azules pastures, forest fragments and continuous rainforest were analyzed, in Los Tuxtlas rainforest fragments of different sizes were analyzed and in Barranca de Metztitlán two types of xerophile scrub with different degrees of disturbance from grazing were analyzed. We assigned dung beetle species to functional groups based on food relocation, beetle size, daily activity period and food preferences, and as measures of functional diversity we used estimates based on multivariate methods. In Montes Azules functional richness was lower in the pastures than in continuous rainforest and rainforest fragments, but fragments and continuous forest include functionally redundant species. In small rainforest fragments (<5 ha) in Los Tuxtlas, dung beetle functional richness was lower than in large rainforest fragments (>20 ha). Functional evenness and functional dispersion did not vary among habitat types or fragment size in these reserves. In contrast, in Metztitlán, functional richness and functional dispersion were different among the vegetation types, but differences were not related to the degree of disturbance by grazing. More redundant species were found in submontane than in crassicaule scrub. For the first time, a decrease in the functional diversity in communities of copro-necrophagous beetles resulting from changes in land use is documented, the potential implications for ecosystem functioning are discussed and a series of variables that could improve the evaluation of functional diversity for this biological group is proposed. PMID:21448292

Effects of logging, hunting, and forest fragment size on physiological stress levels of two sympatric ateline primates in Colombia

PubMed Central

Rimbach, Rebecca; Link, Andrés; Heistermann, Michael; Gómez-Posada, Carolina; Galvis, Nelson; Heymann, Eckhard W.

2013-01-01

Habitat fragmentation and anthropogenic disturbances are of major concern to the conservation of endangered species because of their potentially negative impact on animal populations. Both processes can impose physiological stress (i.e. increased glucocorticoid output) on animals, and chronically elevated stress levels can have detrimental effects on the long-term viability of animal populations. Here, we investigated the effect of fragment size and human impact (logging and hunting pressure) on glucocorticoid levels of two sympatric Neotropical primates, the red howler monkey (Alouatta seniculus) and the critically endangered brown spider monkey (Ateles hybridus). These two species have been reported to contrast strongly in their ability to cope with anthropogenic disturbances. We collected faecal samples from eight spider monkey groups and 31 howler monkey groups, living in seven and 10 different forest fragments in Colombia, respectively. We measured faecal glucocorticoid metabolite (FGCM) levels in both species using previously validated methods. Surprisingly, fragment size did not influence FGCM levels in either species. Spider monkeys showed elevated FGCMs in fragments with the highest level of human impact, whereas we did not find this effect in howler monkeys. This suggests that the two species differ in their physiological responsiveness to anthropogenic changes, further emphasizing why brown spider monkeys are at higher extinction risk than red howler monkeys. If these anthropogenic disturbances persist in the long term, elevated FGCM levels can potentially lead to a state of chronic stress, which might limit the future viability of populations. We propose that FGCM measurements should be used as a tool to monitor populations living in disturbed areas and to assess the success of conservation strategies, such as corridors connecting forest fragments. PMID:27293615

Negative impacts of human land use on dung beetle functional diversity.

PubMed

Barragán, Felipe; Moreno, Claudia E; Escobar, Federico; Halffter, Gonzalo; Navarrete, Dario

2011-03-23

The loss of biodiversity caused by human activity is assumed to alter ecosystem functioning. However our understanding of the magnitude of the effect of these changes on functional diversity and their impact on the dynamics of ecological processes is still limited. We analyzed the functional diversity of copro-necrophagous beetles under different conditions of land use in three Mexican biosphere reserves. In Montes Azules pastures, forest fragments and continuous rainforest were analyzed, in Los Tuxtlas rainforest fragments of different sizes were analyzed and in Barranca de Metztitlán two types of xerophile scrub with different degrees of disturbance from grazing were analyzed. We assigned dung beetle species to functional groups based on food relocation, beetle size, daily activity period and food preferences, and as measures of functional diversity we used estimates based on multivariate methods. In Montes Azules functional richness was lower in the pastures than in continuous rainforest and rainforest fragments, but fragments and continuous forest include functionally redundant species. In small rainforest fragments (<5 ha) in Los Tuxtlas, dung beetle functional richness was lower than in large rainforest fragments (>20 ha). Functional evenness and functional dispersion did not vary among habitat types or fragment size in these reserves. In contrast, in Metztitlán, functional richness and functional dispersion were different among the vegetation types, but differences were not related to the degree of disturbance by grazing. More redundant species were found in submontane than in crassicaule scrub. For the first time, a decrease in the functional diversity in communities of copro-necrophagous beetles resulting from changes in land use is documented, the potential implications for ecosystem functioning are discussed and a series of variables that could improve the evaluation of functional diversity for this biological group is proposed.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stershic, Andrew J.; Dolbow, John E.; Moës, Nicolas

The Thick Level-Set (TLS) model is implemented to simulate brittle media undergoing dynamic fragmentation. This non-local model is discretized by the finite element method with damage represented as a continuous field over the domain. A level-set function defines the extent and severity of damage, and a length scale is introduced to limit the damage gradient. Numerical studies in one dimension demonstrate that the proposed method reproduces the rate-dependent energy dissipation and fragment length observations from analytical, numerical, and experimental approaches. In conclusion, additional studies emphasize the importance of appropriate bulk constitutive models and sufficient spatial resolution of the length scale.

Genotoxic effects of occupational exposure to benzene in gasoline station workers

PubMed Central

SALEM, Eman; EL-GARAWANI, Islam; ALLAM, Heba; EL-AAL, Bahiga Abd; HEGAZY, Mofrih

2017-01-01

Benzene, a hazardous component of gasoline, is a genotoxic class I human carcinogen. This study evaluated the genotoxic effects of occupational exposure to benzene in gasoline stations. Genotoxicity of exposure to benzene was assessed in peripheral blood leucocytes of 62 gasoline station workers and compared with an equal numbers of matched controls using total genomic DNA fragmentation, micronucleus test and cell viability test. An ambient air samples were collected and analyzed for Monitoring of benzene, toluene, ethyl benzene and xylene (BTEX) in work environment and control areas. DNA fragmentation, micronucleus and dead cells percent were significantly higher in exposed workers than controls. Level of benzene, Toluene, Ethyl benzene and xylene in the work environment were higher than the control areas and the permissible limits. Gasoline station workers occupationally exposed to benzene are susceptible to genotoxic effects indicated by increased DNA fragmentation, higher frequency of micronucleus and decreased leukocytes viability. PMID:29070767

Arthroscopic fixation of the clavicle shaft fracture.

PubMed

Kim, Yang-Soo; Lee, Hyo-Jin; Kim, Jong-Ick; Yang, Hyo; Jin, Hong-Ki; Patel, Hiren Kirtibhai; Kim, Jong-Ho; Park, In

2017-01-01

This article describes an arthroscopic technique for the fixation of clavicle shaft fractures. A viewing portal is made 2 cm anterior to the fracture site, and a working portal is made 2 cm lateral to the fracture site. The guide wire for a 4.0-mm cannulated screw is inserted through the fracture site to the medial fracture fragment under arthroscopic guidance. Through the medial fragment, the guide wire is delivered through the skin anteriorly. The fracture is reduced, and then, the guide wire is drilled back across the fracture site to the lateral fracture fragment. After confirming the reduction under arthroscopy, the appropriately sized cannulated screw is inserted after reaming. This arthroscopic technique would be useful for the precise reduction and minimal invasive fixation of clavicle shaft fractures. Preliminary results are encouraging, and further studies with long-term follow-up are needed to determine the precise indications and limitations of the procedure.

DtaRefinery: a software tool for elimination of systematic errors from parent ion mass measurements in tandem mass spectra datasets

DOE Office of Scientific and Technical Information (OSTI.GOV)

Petyuk, Vladislav A.; Mayampurath, Anoop M.; Monroe, Matthew E.

2009-12-16

Hybrid two-stage mass spectrometers capable of both highly accurate mass measurement and MS/MS fragmentation have become widely available in recent years and have allowed for sig-nificantly better discrimination between true and false MS/MS pep-tide identifications by applying relatively narrow windows for maxi-mum allowable deviations for parent ion mass measurements. To fully gain the advantage of highly accurate parent ion mass meas-urements, it is important to limit systematic mass measurement errors. The DtaRefinery software tool can correct systematic errors in parent ion masses by reading a set of fragmentation spectra, searching for MS/MS peptide identifications, then fitting a model that canmore » estimate systematic errors, and removing them. This results in a new fragmentation spectrum file with updated parent ion masses.« less

Chemical Understanding of the Limited Site-Specificity in Molecular Inner-Shell Photofragmentation

DOE PAGES

Inhester, Ludger; Oostenrijk, Bart; Patanen, Minna; ...

2018-02-14

In many cases fragmentation of molecules upon inner-shell ionization is very unspecific with respect to the initially localized ionization site. Often this finding is interpreted in terms of an equilibration of internal energy into vibrational degrees of freedom after Auger decay. In this paper, we investigate the X-ray photofragmentation of ethyl trifluoroacetate upon core electron ionization at environmentally distinct carbon sites using photoelectron–photoion–photoion coincidence measurements and ab initio electronic structure calculations. For all four carbon ionization sites, the Auger decay weakens the same bonds and transfers the two charges to opposite ends of the molecule, which leads to a rapidmore » dissociation into three fragments, followed by further fragmentation steps. Finally, the lack of site specificity is attributed to the character of the dicationic electronic states after Auger decay instead of a fast equilibration of internal energy.« less

Genotoxic effects of occupational exposure to benzene in gasoline station workers.

PubMed

Salem, Eman; El-Garawani, Islam; Allam, Heba; El-Aal, Bahiga Abd; Hegazy, Mofrih

2018-04-07

Benzene, a hazardous component of gasoline, is a genotoxic class I human carcinogen. This study evaluated the genotoxic effects of occupational exposure to benzene in gasoline stations. Genotoxicity of exposure to benzene was assessed in peripheral blood leucocytes of 62 gasoline station workers and compared with an equal numbers of matched controls using total genomic DNA fragmentation, micronucleus test and cell viability test. An ambient air samples were collected and analyzed for Monitoring of benzene, toluene, ethyl benzene and xylene (BTEX) in work environment and control areas. DNA fragmentation, micronucleus and dead cells percent were significantly higher in exposed workers than controls. Level of benzene, Toluene, Ethyl benzene and xylene in the work environment were higher than the control areas and the permissible limits. Gasoline station workers occupationally exposed to benzene are susceptible to genotoxic effects indicated by increased DNA fragmentation, higher frequency of micronucleus and decreased leukocytes viability.

Effects of habitat fragmentation and disturbance on howler monkeys: a review.

PubMed

Arroyo-Rodríguez, Víctor; Dias, Pedro Américo D

2010-01-01

We examined the literature on the effects of habitat fragmentation and disturbance on howler monkeys (genus Alouatta) to (1) identify different threats that may affect howlers in fragmented landscapes; (2) review specific predictions developed in fragmentation theory and (3) identify the empirical evidence supporting these predictions. Although howlers are known for their ability to persist in both conserved and disturbed conditions, we found evidence that they are negatively affected by high levels of habitat loss, fragmentation and degradation. Patch size appears to be the main factor constraining populations in fragmented habitats, probably because patch size is positively related to food availability, and negatively related to anthropogenic pressures, physiological stress and parasite loads. Patch isolation is not a strong predictor of either patch occupancy or population size in howlers, a result that may be related to the ability of howlers to move among forest patches. Thus, we propose that it is probable that habitat loss has larger consistent negative effects on howler populations than habitat fragmentation per se. In general, food availability decreases with patch size, not only due to habitat loss, but also because the density of big trees, plant species richness and howlers' home range size are lower in smaller patches, where howlers' population densities are commonly higher. However, it is unclear which vegetation attributes have the biggest influence on howler populations. Similarly, our knowledge is still limited concerning the effects of postfragmentation threats (e.g. hunting and logging) on howlers living in forest patches, and how several endogenous threats (e.g. genetic diversity, physiological stress, and parasitism) affect the distribution, population structure and persistence of howlers. More long-term studies with comparable methods are necessary to quantify some of the patterns discussed in this review, and determine through meta-analyses whether there are significant inter-specific differences in species' responses to habitat loss and fragmentation. (c) 2009 Wiley-Liss, Inc.

Does the optimal position of the acetabular fragment should be within the radiological normal range for all developmental dysplasia of the hip? A patient-specific finite element analysis.

PubMed

Wang, Xuyi; Peng, Jianping; Li, De; Zhang, Linlin; Wang, Hui; Jiang, Leisheng; Chen, Xiaodong

2016-10-04

The success of Bernese periacetabular osteotomy depends significantly on how extent the acetabular fragment can be corrected to its optimal position. This study was undertaken to investigate whether correcting the acetabular fragment into the so-called radiological "normal" range is the best choice for all developmental dysplasia of the hip with different severities of dysplasia from the biomechanical view? If not, is there any correlation between the biomechanically optimal position of the acetabular fragment and the severity of dysplasia? Four finite element models with different severities of dysplasia were developed. The virtual periacetabular osteotomy was performed with the acetabular fragment rotated anterolaterally to incremental center-edge angles; then, the contact area and pressure and von Mises stress in the cartilage were calculated at different correction angles. The optimal position of the acetabular fragment for patients 1, 2, and 3 was when the acetabular fragment rotated 17° laterally (with the lateral center-edge angle of 36° and anterior center-edge angle of 58°; both were slightly larger than the "normal" range), 25° laterally following further 5° anterior rotation (with the lateral center-edge angle of 31° and anterior center-edge angle of 51°; both were within the "normal" range), and 30° laterally following further 10° anterior rotation (with the lateral center-edge angle of 25° and anterior center-edge angle of 40°; both were less than the "normal" range), respectively. The optimal corrective position of the acetabular fragment is severity dependent rather than within the radiological "normal" range for developmental dysplasia of the hip. We prudently proposed that the optimal correction center-edge angle of mild, moderate, and severe developmental dysplasia of the hip is slightly larger than the "normal" range, within the "normal" range, and less than the lower limit of the "normal" range, respectively.

Mycobacterium tuberculosis Cell Wall Fragments Released upon Bacterial Contact with the Human Lung Mucosa Alter the Neutrophil Response to Infection

PubMed Central

Scordo, Julia M.; Arcos, Jesús; Kelley, Holden V.; Diangelo, Lauren; Sasindran, Smitha J.; Youngmin, Ellie; Wewers, Mark D.; Wang, Shu-Hua; Balada-Llasat, Joan-Miquel; Torrelles, Jordi B.

2017-01-01

In 2016, the World Health Organization reported that one person dies of tuberculosis (TB) every 21 s. A host environment that Mycobacterium tuberculosis (M.tb) finds during its route of infection is the lung mucosa bathing the alveolar space located in the deepest regions of the lungs. We published that human lung mucosa, or alveolar lining fluid (ALF), contains an array of hydrolytic enzymes that can significantly alter the M.tb surface during infection by cleaving off parts of its cell wall. This interaction results in two different outcomes: modifications on the M.tb cell wall surface and release of M.tb cell wall fragments into the environment. Typically, one of the first host immune cells at the site of M.tb infection is the neutrophil. Neutrophils can mount an extracellular and intracellular innate immune response to M.tb during infection. We hypothesized that exposure of neutrophils to ALF-induced M.tb released cell wall fragments would prime neutrophils to control M.tb infection better. Our results show that ALF fragments activate neutrophils leading to an increased production of inflammatory cytokines and oxidative radicals. However, neutrophil exposure to these fragments reduces production of chemoattractants (i.e., interleukin-8), and degranulation, with the subsequent reduction of myeloperoxidase release, and does not induce cytotoxicity. Unexpectedly, these ALF fragment-derived modulations in neutrophil activity do not further, either positively or negatively, contribute to the intracellular control of M.tb growth during infection. However, secreted products from neutrophils primed with ALF fragments are capable of regulating the activity of resting macrophages. These results indicate that ALF-induced M.tb fragments could further contribute to the control of M.tb growth and local killing by resident neutrophils by switching on the total oxidative response and limiting migration of neutrophils to the infection site. PMID:28373877

Mycobacterium tuberculosis Cell Wall Fragments Released upon Bacterial Contact with the Human Lung Mucosa Alter the Neutrophil Response to Infection.

PubMed

Scordo, Julia M; Arcos, Jesús; Kelley, Holden V; Diangelo, Lauren; Sasindran, Smitha J; Youngmin, Ellie; Wewers, Mark D; Wang, Shu-Hua; Balada-Llasat, Joan-Miquel; Torrelles, Jordi B

2017-01-01

In 2016, the World Health Organization reported that one person dies of tuberculosis (TB) every 21 s. A host environment that Mycobacterium tuberculosis ( M.tb ) finds during its route of infection is the lung mucosa bathing the alveolar space located in the deepest regions of the lungs. We published that human lung mucosa, or alveolar lining fluid (ALF), contains an array of hydrolytic enzymes that can significantly alter the M.tb surface during infection by cleaving off parts of its cell wall. This interaction results in two different outcomes: modifications on the M.tb cell wall surface and release of M.tb cell wall fragments into the environment. Typically, one of the first host immune cells at the site of M.tb infection is the neutrophil. Neutrophils can mount an extracellular and intracellular innate immune response to M.tb during infection. We hypothesized that exposure of neutrophils to ALF-induced M.tb released cell wall fragments would prime neutrophils to control M.tb infection better. Our results show that ALF fragments activate neutrophils leading to an increased production of inflammatory cytokines and oxidative radicals. However, neutrophil exposure to these fragments reduces production of chemoattractants (i.e., interleukin-8), and degranulation, with the subsequent reduction of myeloperoxidase release, and does not induce cytotoxicity. Unexpectedly, these ALF fragment-derived modulations in neutrophil activity do not further, either positively or negatively, contribute to the intracellular control of M.tb growth during infection. However, secreted products from neutrophils primed with ALF fragments are capable of regulating the activity of resting macrophages. These results indicate that ALF-induced M.tb fragments could further contribute to the control of M.tb growth and local killing by resident neutrophils by switching on the total oxidative response and limiting migration of neutrophils to the infection site.

Evolution of Particle Size Distributions in Fragmentation Over Time

NASA Astrophysics Data System (ADS)

Charalambous, C. A.; Pike, W. T.

2013-12-01

We present a new model of fragmentation based on a probabilistic calculation of the repeated fracture of a particle population. The resulting continuous solution, which is in closed form, gives the evolution of fragmentation products from an initial block, through a scale-invariant power-law relationship to a final comminuted powder. Models for the fragmentation of particles have been developed separately in mainly two different disciplines: the continuous integro-differential equations of batch mineral grinding (Reid, 1965) and the fractal analysis of geophysics (Turcotte, 1986) based on a discrete model with a single probability of fracture. The first gives a time-dependent development of the particle-size distribution, but has resisted a closed-form solution, while the latter leads to the scale-invariant power laws, but with no time dependence. Bird (2009) recently introduced a bridge between these two approaches with a step-wise iterative calculation of the fragmentation products. The development of the particle-size distribution occurs with discrete steps: during each fragmentation event, the particles will repeatedly fracture probabilistically, cascading down the length scales to a final size distribution reached after all particles have failed to further fragment. We have identified this process as the equivalent to a sequence of trials for each particle with a fixed probability of fragmentation. Although the resulting distribution is discrete, it can be reformulated as a continuous distribution in maturity over time and particle size. In our model, Turcotte's power-law distribution emerges at a unique maturation index that defines a regime boundary. Up to this index, the fragmentation is in an erosional regime with the initial particle size setting the scaling. Fragmentation beyond this index is in a regime of comminution with rebreakage of the particles down to the size limit of fracture. The maturation index can increment continuously, for example under grinding conditions, or as discrete steps, such as with impact events. In both cases our model gives the energy associated with the fragmentation in terms of the developing surface area of the population. We show the agreement of our model to the evolution of particle size distributions associated with episodic and continuous fragmentation and how the evolution of some popular fractals may be represented using this approach. C. A. Charalambous and W. T. Pike (2013). Multi-Scale Particle Size Distributions of Mars, Moon and Itokawa based on a time-maturation dependent fragmentation model. Abstract Submitted to the AGU 46th Fall Meeting. Bird, N. R. A., Watts, C. W., Tarquis, A. M., & Whitmore, A. P. (2009). Modeling dynamic fragmentation of soil. Vadose Zone Journal, 8(1), 197-201. Reid, K. J. (1965). A solution to the batch grinding equation. Chemical Engineering Science, 20(11), 953-963. Turcotte, D. L. (1986). Fractals and fragmentation. Journal of Geophysical Research: Solid Earth 91(B2), 1921-1926.

Topography of succinate dehydrogenase in the mitochondrial inner membrane. A study using limited proteolysis and immunoblotting.

PubMed Central

Clarkson, G H; Neagle, J; Lindsay, J G

1991-01-01

The arrangement of the large (70,000-Mr) and small (30,000-Mr) subunits of succinate dehydrogenase in the mitochondrial inner membrane was investigated by immunoblot analysis of bovine heart mitochondria (right-side-out, outer membrane disrupted) or submitochondrial particles (inside-out) that had been subjected to surface-specific proteolysis. Both subunits were resistant to proteinase treatment provided that the integrity of the inner membrane was preserved, suggesting that neither subunit is exposed at the cytoplasmic surface of the membrane. The bulk of the small subunit appears to protrude into the matrix compartment, since the 30,000-Mr polypeptide is degraded extensively during limited proteolysis of submitochondrial particles without the appearance of an immunologically reactive membrane-associated fragment: moreover, a soluble 27,000-Mr peptide derived from this subunit is observed transiently on incubation with trypsin. Similar data obtained from the large subunit suggest that this polypeptide interacts with the matrix side of the inner membrane via two distinct domains; these are detected as stable membrane-associated fragments of 32,000 Mr and 27,000 Mr after treatment of submitochondrial particles with papain or proteinase K, although the 27,000-Mr fragment can be degraded further to low-Mr peptides with trypsin or alpha-chymotrypsin. A stable 32,000-34,000-Mr fragment is generated by a variety of specific and non-specific proteinases, indicating that it may be embedded largely within the lipid bilayer, or is inaccessible to proteolytic attack owing to its proximity to the surface of the intact membrane, possibly interacting with the hydrophobic membrane anchoring polypeptides of the succinate: ubiquinone reductase complex. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. PMID:1996968

Use of a laparoscopic specimen retrieval pouch to facilitate removal of intact or fragmented cystic calculi from standing sedated horses: 8 cases (2012-2015).

PubMed

Katzman, Scott A; Vaughan, Betsy; Nieto, Jorge E; Galuppo, Larry D

2016-08-01

OBJECTIVE To evaluate the use of a laparoscopic specimen retrieval pouch for removal of intact or fragmented cystic calculi from standing horses. DESIGN Retrospective case series. ANIMALS 8 horses (5 geldings and 3 mares) with cystic calculi. PROCEDURES Physical examination and cystoscopic, ultrasonographic, and hematologic evaluations of urinary tract function were performed for each horse. A diagnosis of cystic calculus was made on the basis of results of cystoscopy and ultrasonography. Concurrent urolithiasis or other urinary tract abnormalities identified during preoperative evaluation were recorded. Horses were sedated and placed in standing stocks, and the perineum was aseptically prepared. Direct access to the urinary bladder was gained in geldings via perineal urethrotomy or in mares by a transurethral approach. Calculi were visualized endoscopically, manipulated into the retrieval pouch, and removed intact or fragmented (for larger calculi). RESULTS For 4 geldings and 1 mare, fragmentation was necessary to facilitate calculus removal. Mean duration of surgery was 125 minutes, and trauma to the urinary bladder and urethra was limited to areas of hyperemia and submucosal petechiation. No postoperative complications were encountered for any horse. When lithotripsy was required, the retrieval pouch provided an effective means of stabilizing calculi and containing the fragments for removal. CONCLUSIONS AND CLINICAL RELEVANCE Use of the laparoscopic specimen retrieval pouch was an effective, minimally traumatic method for retrieving cystic calculi from standing horses. The pouch protected the urinary bladder and urethra from trauma during calculus removal and allowed for stabilization, containment, and fragmentation of calculi when necessary.

Hyperthermal (1-100 eV) nitrogen ion scattering damage to D-ribose and 2-deoxy-D-ribose films

DOE Office of Scientific and Technical Information (OSTI.GOV)

Deng Zongwu; Bald, Ilko; Illenberger, Eugen

2007-10-14

Highly charged heavy ion traversal of a biological medium can produce energetic secondary fragment ions. These fragment ions can in turn cause collisional and reactive scattering damage to DNA. Here we report hyperthermal (1-100 eV) scattering of one such fragment ion (N{sup +}) from biologically relevant sugar molecules D-ribose and 2-deoxy-D-ribose condensed on polycrystalline Pt substrate. The results indicate that N{sup +} ion scattering at kinetic energies down to 10 eV induces effective decomposition of both sugar molecules and leads to the desorption of abundant cation and anion fragments. Use of isotope-labeled molecules (5-{sup 13}C D-ribose and 1-D D-ribose) partlymore » reveals some site specificity of the fragment origin. Several scattering reactions are also observed. Both ionic and neutral nitrogen atoms abstract carbon from the molecules to form CN{sup -} anion at energies down to {approx}5 eV. N{sup +} ions also abstract hydrogen from hydroxyl groups of the molecules to form NH{sup -} and NH{sub 2}{sup -} anions. A fraction of O/O{sup -} fragments abstract hydrogen to form OH{sup -}. The formation of H{sub 3}O{sup +} ions also involves hydrogen abstraction as well as intramolecular proton transfer. These findings suggest a variety of severe damaging pathways to DNA molecules which occur on the picosecond time scale following heavy ion irradiation of a cell, and prior to the late diffusion-limited homogeneous chemical processes.« less

Determination of patulin in apple juice by high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry.

PubMed

Sewram, V; Nair, J J; Nieuwoudt, T W; Leggott, N L; Shephard, G S

2000-11-03

An HPLC-MS-MS method with selected reaction monitoring (SRM) for the determination of patulin in apple juice samples is described. Mass spectrometric detection was accomplished following atmospheric pressure chemical ionization (APCI) in both positive and negative ion modes. Collision induced dissociation (CID) of the protonated molecular ion led initially to the loss of H2O (fragment m/z 137). At higher energies CO is lost from both the protonated parent molecule (fragment m/z 127) and the dehydrated molecular ion (fragment m/z 109). In contrast, CID of the deprotonated molecular ion led initially to the fragment at m/z 109 corresponding to the loss of either CO2 or acetaldehyde, followed at higher CID energy by the loss of H2O (fragment m/z 135) and CO (fragment m/z 125) from the deprotonated molecular ion. Detection in the negative ion mode proved superior and a linear response was observed over the injected range from 6 to 200 ng patulin. Apple juice samples spiked with patulin between 10 and 135 microg/l were analyzed following liquid-liquid extraction with ethyl acetate and clean up with sodium carbonate. Utilizing reversed-phase HPLC with acetonitrile-water (10:90) at 0.5 ml/min, levels down to 10 microg/l were readily quantified and a detection limit of 4 microg/l was attainable at a signal-to-noise (SIN) ratio of 4. The MS data for the spiked samples compared well to the UV data and when plotted against each other displayed a correlation coefficient (R) of 0.99.

The relative effects of habitat loss and fragmentation on population genetic variation in the red-cockaded woodpecker (Picoides borealis).

PubMed

Bruggeman, Douglas J; Wiegand, Thorsten; Fernández, Néstor

2010-09-01

The relative influence of habitat loss, fragmentation and matrix heterogeneity on the viability of populations is a critical area of conservation research that remains unresolved. Using simulation modelling, we provide an analysis of the influence both patch size and patch isolation have on abundance, effective population size (N(e)) and F(ST). An individual-based, spatially explicit population model based on 15 years of field work on the red-cockaded woodpecker (Picoides borealis) was applied to different landscape configurations. The variation in landscape patterns was summarized using spatial statistics based on O-ring statistics. By regressing demographic and genetics attributes that emerged across the landscape treatments against proportion of total habitat and O-ring statistics, we show that O-ring statistics provide an explicit link between population processes, habitat area, and critical thresholds of fragmentation that affect those processes. Spatial distances among land cover classes that affect biological processes translated into critical scales at which the measures of landscape structure correlated best with genetic indices. Therefore our study infers pattern from process, which contrasts with past studies of landscape genetics. We found that population genetic structure was more strongly affected by fragmentation than population size, which suggests that examining only population size may limit recognition of fragmentation effects that erode genetic variation. If effective population size is used to set recovery goals for endangered species, then habitat fragmentation effects may be sufficiently strong to prevent evaluation of recovery based on the ratio of census:effective population size alone.

SKI2 mediates degradation of RISC 5′-cleavage fragments and prevents secondary siRNA production from miRNA targets in Arabidopsis

PubMed Central

Branscheid, Anja; Marchais, Antonin; Schott, Gregory; Lange, Heike; Gagliardi, Dominique; Andersen, Stig Uggerhøj; Voinnet, Olivier; Brodersen, Peter

2015-01-01

Small regulatory RNAs are fundamental in eukaryotic and prokaryotic gene regulation. In plants, an important element of post-transcriptional control is effected by 20–24 nt microRNAs (miRNAs) and short interfering RNAs (siRNAs) bound to the ARGONAUTE1 (AGO1) protein in an RNA induced silencing complex (RISC). AGO1 may cleave target mRNAs with small RNA complementarity, but the fate of the resulting cleavage fragments remains incompletely understood. Here, we show that SKI2, SKI3 and SKI8, subunits of a cytoplasmic cofactor of the RNA exosome, are required for degradation of RISC 5′, but not 3′-cleavage fragments in Arabidopsis. In the absence of SKI2 activity, many miRNA targets produce siRNAs via the RNA-dependent RNA polymerase 6 (RDR6) pathway. These siRNAs are low-abundant, and map close to the cleavage site. In most cases, siRNAs were produced 5′ to the cleavage site, but several examples of 3′-spreading were also identified. These observations suggest that siRNAs do not simply derive from RDR6 action on stable 5′-cleavage fragments and hence that SKI2 has a direct role in limiting secondary siRNA production in addition to its function in mediating degradation of 5′-cleavage fragments. PMID:26464441

[Preparation of monoclonal antibody against 4-amylphenol and homology modeling of its Fv fragment].

PubMed

Cheng, Lei; Wu, Haizhen; Fei, Jing; Zhang, Lujia; Ye, Jiang; Zhang, Huizhan

2017-03-01

Objective To prepare and characterize a monoclonal antibody (mAb) against 4-amylphenol (4-AP), clone its cDNA sequence and make homology modeling for its Fv fragment. Methods A high-affinity anti-4-AP mAb was generated from a hybridoma cell line F10 using electrofusion between splenocytes from APA-BSA-immunized mouse and Sp2/0 myeloma cells. Then we extracted the mRNA of F10 cells and cloned the cDNA of mAb. The homology modeling and molecular docking of its Fv fragment was conducted with biological software. Results Under the optimum conditions, the ic-ELISA equation was y=A 2 +(A 1 -A 2 )/(1+(x/x 0 ) p ) (A 1 =1.28; A 2 =-0.066; x 0 =12560.75; p=0.74) with a correlation coefficient (R 2 ) of 0.997. The lowest detectable limit was 0.65 μg/mL. The heavy and light chains of mAb respectively belonged to IgG1 and Kappa. The homology modeling and molecular docking studies revealed that the binding of 4-Ap and mAb was attributed to the hydrogen bond and hydrophobic interactions. Conclusion The study successfully established a stable 4-AP mAb-secreting hybridoma cell line. The study on spatial structure of Fv fragment using homology modeling provided a reference for the development and design of single chain variable fragments.

LipidFrag: Improving reliability of in silico fragmentation of lipids and application to the Caenorhabditis elegans lipidome

PubMed Central

Neumann, Steffen; Schmitt-Kopplin, Philippe

2017-01-01

Lipid identification is a major bottleneck in high-throughput lipidomics studies. However, tools for the analysis of lipid tandem MS spectra are rather limited. While the comparison against spectra in reference libraries is one of the preferred methods, these libraries are far from being complete. In order to improve identification rates, the in silico fragmentation tool MetFrag was combined with Lipid Maps and lipid-class specific classifiers which calculate probabilities for lipid class assignments. The resulting LipidFrag workflow was trained and evaluated on different commercially available lipid standard materials, measured with data dependent UPLC-Q-ToF-MS/MS acquisition. The automatic analysis was compared against manual MS/MS spectra interpretation. With the lipid class specific models, identification of the true positives was improved especially for cases where candidate lipids from different lipid classes had similar MetFrag scores by removing up to 56% of false positive results. This LipidFrag approach was then applied to MS/MS spectra of lipid extracts of the nematode Caenorhabditis elegans. Fragments explained by LipidFrag match known fragmentation pathways, e.g., neutral losses of lipid headgroups and fatty acid side chain fragments. Based on prediction models trained on standard lipid materials, high probabilities for correct annotations were achieved, which makes LipidFrag a good choice for automated lipid data analysis and reliability testing of lipid identifications. PMID:28278196

The effects of landscape fragmentation on pollination dynamics: absence of evidence not evidence of absence.

PubMed

Hadley, Adam S; Betts, Matthew G

2012-08-01

Animal-mediated pollination is essential for both ecosystem services and conservation of global biodiversity, but a growing body of work reveals that it is negatively affected by anthropogenic disturbance. Landscape-scale disturbance results in two often inter-related processes: (1) habitat loss, (2) disruptions of habitat configuration (i.e. fragmentation). Understanding the relative effects of such processes is critical in designing effective management strategies to limit pollination and pollinator decline. We reviewed existing published work from 1989 to 2009 and found that only six of 303 studies considering the influence of landscape context on pollination separated the effects of habitat loss from fragmentation. We provide a synthesis of the current landscape, behavioural, and pollination ecology literature in order to present preliminary multiple working hypotheses explaining how these two landscape processes might independently influence pollination dynamics. Landscape disturbance primarily influences three components of pollination interactions: pollinator density, movement, and plant demography. We argue that effects of habitat loss on each of these components are likely to differ substantially from the effects of fragmentation, which is likely to be more complex and may influence each pollination component in contrasting ways. The interdependency between plants and animals inherent to pollination systems also has the possibility to drive cumulative effects of fragmentation, initiating negative feedback loops between animals and the plants they pollinate. Alternatively, due to their asymmetrical structure, pollination networks may be relatively robust to fragmentation. Despite the potential importance of independent effects of habitat fragmentation, its effects on pollination remain largely untested. We postulate that variation across studies in the effects of 'fragmentation' owes much to artifacts of the sampling regimes adopted, particularly (1) incorrectly separating fragmentation from habitat loss, and (2) mis-matches in spatial scale between landscapes studied and the ecological processes of interest. The field of landscape pollination ecology could be greatly advanced through the consideration and quantification of the matrix, landscape functional connectivity, and pollinator movement behaviour in response to these elements. Studies designed to disentangle the independent effects of habitat loss and fragmentation are essential for gaining insight into landscape-mediated pollination declines, implementing effective conservation measures, and optimizing ecosystem services in complex landscapes. © 2011 The Authors. Biological Reviews © 2011 Cambridge Philosophical Society.

Nucleotide exchange and excision technology DNA shuffling and directed evolution.

PubMed

Speck, Janina; Stebel, Sabine C; Arndt, Katja M; Müller, Kristian M

2011-01-01

Remarkable success in optimizing complex properties within DNA and proteins has been achieved by directed evolution. In contrast to various random mutagenesis methods and high-throughput selection methods, the number of available DNA shuffling procedures is limited, and protocols are often difficult to adjust. The strength of the nucleotide exchange and excision technology (NExT) DNA shuffling described here is the robust, efficient, and easily controllable DNA fragmentation step based on random incorporation of the so-called 'exchange nucleotides' by PCR. The exchange nucleotides are removed enzymatically, followed by chemical cleavage of the DNA backbone. The oligonucleotide pool is reassembled into full-length genes by internal primer extension, and the recombined gene library is amplified by standard PCR. The technique has been demonstrated by shuffling a defined gene library of chloramphenicol acetyltransferase variants using uridine as fragmentation defining exchange nucleotide. Substituting 33% of the dTTP with dUTP in the incorporation PCR resulted in shuffled clones with an average parental fragment size of 86 bases and revealed a mutation rate of only 0.1%. Additionally, a computer program (NExTProg) has been developed that predicts the fragment size distribution depending on the relative amount of the exchange nucleotide.

High voltage fragmentation of composites from secondary raw materials - Potential and limitations.

PubMed

Leißner, T; Hamann, D; Wuschke, L; Jäckel, H-G; Peuker, U A

2018-04-01

The comminution of composites for liberation of valuable components is a costly and energy-intensive process within the recycling of spent products. It therefore is continuously studied and optimized. In addition to conventional mechanical comminution innovative new principles for size reduction have been developed. One is the use of high voltage (HV) pulses, which is known to be a technology selectively liberating along phase boundaries. This technology offers the advantage of targeted liberation, preventing overgrinding of the material and thus improving the overall processing as well as product quality. In this study, the high voltage fragmentation of three different non-brittle composites (galvanized plastics, carbon fibre composites, electrode foils from Li-ion batteries) was investigated. The influence of pulse rate, number of pulses and filling level on the liberation and efficiency of comminution is discussed. Using the guideline VDI 2225 HV, fragmentation is compared to conventional mechanical comminution with respect to numerous criteria such as cost, throughput, energy consumption, availability and scalability. It was found that at current state of development, HV fragmentation cannot compete with mechanical comminution beyond laboratory scale. Copyright © 2017 Elsevier Ltd. All rights reserved.

How fragmentation and corridors affect wind dynamics and seed dispersal in open habitats

PubMed Central

Damschen, Ellen I.; Baker, Dirk V.; Bohrer, Gil; Nathan, Ran; Orrock, John L.; Turner, Jay R.; Brudvig, Lars A.; Haddad, Nick M.; Levey, Douglas J.; Tewksbury, Joshua J.

2014-01-01

Determining how widespread human-induced changes such as habitat loss, landscape fragmentation, and climate instability affect populations, communities, and ecosystems is one of the most pressing environmental challenges. Critical to this challenge is understanding how these changes are affecting the movement abilities and dispersal trajectories of organisms and what role conservation planning can play in promoting movement among remaining fragments of suitable habitat. Whereas evidence is mounting for how conservation strategies such as corridors impact animal movement, virtually nothing is known for species dispersed by wind, which are often mistakenly assumed to not be limited by dispersal. Here, we combine mechanistic dispersal models, wind measurements, and seed releases in a large-scale experimental landscape to show that habitat corridors affect wind dynamics and seed dispersal by redirecting and bellowing airflow and by increasing the likelihood of seed uplift. Wind direction interacts with landscape orientation to determine when corridors provide connectivity. Our results predict positive impacts of connectivity and patch shape on species richness of wind-dispersed plants, which we empirically illustrate using 12 y of data from our experimental landscapes. We conclude that habitat fragmentation and corridors strongly impact the movement of wind-dispersed species, which has community-level consequences. PMID:24567398

Genetic variation and genetic structure of the endangered species Sinowilsonia henryi Hemsi. (Hamamelidaceae) revealed by amplified fragment length polymorphism (AFLP) markers.

PubMed

Zhang, H; Ji, W L; Li, M; Zhou, L Y

2015-10-14

Comprehensive research of genetic variation is crucial in designing conservation strategies for endangered and threatened species. Sinowilsonia henryi Hemsi. is a tertiary relic with a limited geographical distribution in the central and western areas of China. It is endangered because of climate change and habitat fragmentation over the last thousands of years. In this study, amplified fragment length polymorphism markers were utilized to estimate genetic diversity and genetic structure in and among S. henryi. In this study, Nei's genetic diversity and Shannon's information index were found to be 0.192 and 0.325 respectively, indicating a moderate-to-high genetic diversity in species. According to analysis of molecular variation results, 32% of the genetic variation was shown to be partitioned among populations, demonstrating a relatively high genetic divergence; this was supported by principal coordinate analysis and unweighted pair-group method with arithmetic average analysis. Moreover, the Mantel test showed that there was no significant correlation between genetic and geographical distances. The above results can be explained by the effects of habitat fragmentation, history traits, and gene drift. Based on the results, several implications were indicated and suggestions proposed for preservation strategies for this species.

Piecing together the fragments: Elucidating edge effects on forest carbon dynamics

NASA Astrophysics Data System (ADS)

Hutyra, L.; Smith, I. A.; Reinmann, A.; Marrs, J.; Thompson, J.

2017-12-01

Forest fragmentation is pervasive throughout the world's forests, impacting growing conditions and carbon dynamics through edge effects that produce gradients in microclimate, biogeochemistry, and stand structure. Despite the majority of the world's forests being <1km from an edge, our understanding of forest carbon dynamics is largely derived from intact forest systems. In the northeastern USA, we find that over 23% of the current forest area is just 30m from an agricultural or developed edge. Edge effects on the carbon cycle vary in their magnitude by biome, but current forest carbon accounting methods and ecosystem models largely do not include edge effects, highlighting an important gap in our understanding of the terrestrial carbon cycle. Characterizing the role of forest fragmentation in regional and global biogeochemical cycles necessitates advancing our understanding of how shifts in microenvironment at the forest edge interact with local prevailing drivers of global change and limitations to microbial activity and forest growth. This study synthesizes the literature related to edge effects and the carbon cycle, considering how fragmentation affects the growing conditions of the world's remaining forests based on risks and opportunities for forests near the edge.

Marine fisheries declines viewed upside down: human impacts on consumer-driven nutrient recycling.

PubMed

Layman, Craig A; Allgeier, Jacob E; Rosemond, Amy D; Dahlgren, Craig P; Yeager, Lauren A

2011-03-01

We quantified how two human impacts (overfishing and habitat fragmentation) in nearshore marine ecosystems may affect ecosystem function by altering the role of fish as nutrient vectors. We empirically quantified size-specific excretion rates of one of the most abundant fishes (gray snapper, Lutjanus griseus) in The Bahamas and combined these with surveys of fish abundance to estimate population-level excretion rates. The study was conducted across gradients of two human disturbances: overfishing and ecosystem fragmentation (estuaries bisected by roads), to evaluate how each could result in reduced population-level nutrient cycling by consumers. Mean estimated N and P excretion rates for gray snapper populations were on average 456% and 541% higher, respectively, in unfished sites. Ecosystem fragmentation resulted in significant reductions of recycling rates by snapper, with degree of creek fragmentation explaining 86% and 72% of the variance in estimated excretion for dissolved N and P, respectively. Additionally, we used nutrient limitation assays and primary producer nutrient content to provide a simple example of how marine fishery declines may affect primary production. This study provides an initial step toward integrating marine fishery declines and consumer-driven nutrient recycling to more fully understand the implications of human impacts in marine ecosystems.

How fragmentation and corridors affect wind dynamics and seed dispersal in open habitats.

PubMed

Damschen, Ellen I; Baker, Dirk V; Bohrer, Gil; Nathan, Ran; Orrock, John L; Turner, Jay R; Brudvig, Lars A; Haddad, Nick M; Levey, Douglas J; Tewksbury, Joshua J

2014-03-04

Determining how widespread human-induced changes such as habitat loss, landscape fragmentation, and climate instability affect populations, communities, and ecosystems is one of the most pressing environmental challenges. Critical to this challenge is understanding how these changes are affecting the movement abilities and dispersal trajectories of organisms and what role conservation planning can play in promoting movement among remaining fragments of suitable habitat. Whereas evidence is mounting for how conservation strategies such as corridors impact animal movement, virtually nothing is known for species dispersed by wind, which are often mistakenly assumed to not be limited by dispersal. Here, we combine mechanistic dispersal models, wind measurements, and seed releases in a large-scale experimental landscape to show that habitat corridors affect wind dynamics and seed dispersal by redirecting and bellowing airflow and by increasing the likelihood of seed uplift. Wind direction interacts with landscape orientation to determine when corridors provide connectivity. Our results predict positive impacts of connectivity and patch shape on species richness of wind-dispersed plants, which we empirically illustrate using 12 y of data from our experimental landscapes. We conclude that habitat fragmentation and corridors strongly impact the movement of wind-dispersed species, which has community-level consequences.

PubChem atom environments.

PubMed

Hähnke, Volker D; Bolton, Evan E; Bryant, Stephen H

2015-01-01

Atom environments and fragments find wide-spread use in chemical information and cheminformatics. They are the basis of prediction models, an integral part in similarity searching, and employed in structure search techniques. Most of these methods were developed and evaluated on the relatively small sets of chemical structures available at the time. An analysis of fragment distributions representative of most known chemical structures was published in the 1970s using the Chemical Abstracts Service data system. More recently, advances in automated synthesis of chemicals allow millions of chemicals to be synthesized by a single organization. In addition, open chemical databases are readily available containing tens of millions of chemical structures from a multitude of data sources, including chemical vendors, patents, and the scientific literature, making it possible for scientists to readily access most known chemical structures. With this availability of information, one can now address interesting questions, such as: what chemical fragments are known today? How do these fragments compare to earlier studies? How unique are chemical fragments found in chemical structures? For our analysis, after hydrogen suppression, atoms were characterized by atomic number, formal charge, implicit hydrogen count, explicit degree (number of neighbors), valence (bond order sum), and aromaticity. Bonds were differentiated as single, double, triple or aromatic bonds. Atom environments were created in a circular manner focused on a central atom with radii from 0 (atom types) up to 3 (representative of ECFP_6 fragments). In total, combining atom types and atom environments that include up to three spheres of nearest neighbors, our investigation identified 28,462,319 unique fragments in the 46 million structures found in the PubChem Compound database as of January 2013. We could identify several factors inflating the number of environments involving transition metals, with many seemingly due to erroneous interpretation of structures from patent data. Compared to fragmentation statistics published 40 years ago, the exponential growth in chemistry is mirrored in a nearly eightfold increase in the number of unique chemical fragments; however, this result is clearly an upper bound estimate as earlier studies employed structure sampling approaches and this study shows that a relatively high rate of atom fragments are found in only a single chemical structure (singletons). In addition, the percentage of singletons grows as the size of the chemical fragment is increased. The observed growth of the numbers of unique fragments over time suggests that many chemically possible connections of atom types to larger fragments have yet to be explored by chemists. A dramatic drop in the relative rate of increase of atom environments from smaller to larger fragments shows that larger fragments mainly consist of diverse combinations of a limited subset of smaller fragments. This is further supported by the observed concomitant increase of singleton atom environments. Combined, these findings suggest that there is considerable opportunity for chemists to combine known fragments to novel chemical compounds. The comparison of PubChem to an older study of known chemical structures shows noticeable differences. The changes suggest advances in synthetic capabilities of chemists to combine atoms in new patterns. Log-log plots of fragment incidence show small numbers of fragments are found in many structures and that large numbers of fragments are found in very few structures, with nearly half being novel using the methods in this work. The relative decrease in the count of new fragments as a function of size further suggests considerable opportunity for more novel chemicals exists. Lastly, the differences in atom environment diversity between PubChem Substance and Compound showcase the effect of PubChem standardization protocols, but also indicate that a normalization procedure for atom types, functional groups, and tautomeric/resonance forms based on atom environments is possible. The complete sets of atom types and atom environments are supplied as supporting information.

Beneficial effect of directional freezing on in vitro viability of cryopreserved sheep whole ovaries and ovarian cortical slices.

PubMed

Maffei, S; Pennarossa, G; Brevini, T A L; Arav, A; Gandolfi, F

2014-01-01

Does directional freezing improve the structural and functional integrity of ovarian fragments compared with conventional slow freezing and to whole ovary cryopreservation? Compared with slow freezing, the use of directional freezing significantly improves all structural and functional parameters of ovarian fragments assessed in vitro and, overall, whole ovaries were better preserved than ovarian fragments. Directional freezing has been developed to provide an alternative way to cryopreserve large biological samples and it is known to improve the structural and functional integrity of whole ovaries. Conventional slow freezing of ovarian fragments is the procedure more widely used in clinical settings but it causes substantial structural damage that limits the functional period after transfer back into the patient. We performed a 2 × 2 factorial design experiment on a total of 40 sheep ovaries, divided into four groups (n = 10 ovaries per group): (i) directional freezing of whole ovary (DFwo); (ii) directional freezing of ovarian fragments (DFof); (iii) conventional freezing of whole ovary (CFwo); (iv) conventional freezing of ovarian fragments (CFof). An additional eight ovaries were used as fresh controls. Ewe ovaries were randomly assigned to one of the experimental groups and frozen accordingly. Upon thawing, ovarian tissue was examined morphologically and cultured in vitro for 7 days. Samples were analyzed for cell proliferation and apoptosis, for DNA damage and repair activity, and for the presence of a panel of heat shock proteins (HSPs) by immunohistochemistry. Most studied parameters were significantly improved (P < 0.05) in all samples cryopreserved with directional compared with slow freezing. The proportion of primordial follicles, which developed to the primary stage in whole ovaries (53 ± 1.7%) and in ovarian fragments (44 ± 1.8%) cryopreserved with directional freezing, was greater than with slow frozen whole ovaries (6 ± 0.5%, P = 0.001) or fragments (32 ± 1.5%, P = 0.004). After 7 days of culture, cell proliferation in DFwo (28 ± 0.73%) was the highest of all groups (P < 0.05) followed by DFof (23 ± 0.81%), CFof (20 ± 0.79%) and CFwo (9 ± 0.85%). Directional freezing also resulted in a better preservation of the cell capacity to repair DNA damage compared with slow freezing both in whole ovaries and ovarian fragments. Apoptosis and HSP protein levels were significantly increased only in the CFwo group. Direct comparison demonstrated that, overall, DFwo had better parameters than DFof and was no different from the fresh controls. The study is limited to an in vitro evaluation and uses sheep ovaries, which are smaller than human ovaries and therefore may withstand the procedures better. Improved integrity of ovarian morphology may translate to improved outcomes after transplantation. Alternatively, the particularly good preservation of whole ovaries suggests they could provide a source of ovarian follicles for in vitro culture in those cases when the presence of malignant cells poses a substantial risk for the patient. Supported by: Associazione Italiana per la Ricerca sul Cancro (AIRC) IG 10376, Carraresi Foundation and by Legge 7 Regione Autonoma Sardegna (R.A.S). There are no conflicts of interest.

Multichannel processes of H2O in the 18 eV region

NASA Technical Reports Server (NTRS)

Wu, C. Y. Robert; Judge, D. L.

1988-01-01

Measurements were made of: (1) the fluorescence cross sections of OH(A 2Sigma+) fragments; (2) the absolute cross sections producing H atoms in the n = 2, 3, and 4 states; (3) the cross section for producing excited O atoms which has an upper limit of 5 x 10 to the -21 sq cm; and (4) the fluorescence cross section for producing H2(a 3Sigma g +) fragments. It is shown that, in the 16-20 eV region, there are excellent correspondences in the peak positions and spacings among the photoabsorption, photoionization spectra, and fluorescence functions of OH(A) and H(n).

Plant genotyping using fluorescently tagged inter-simple sequence repeats (ISSRs): basic principles and methodology.

PubMed

Prince, Linda M

2015-01-01

Inter-simple sequence repeat PCR (ISSR-PCR) is a fast, inexpensive genotyping technique based on length variation in the regions between microsatellites. The method requires no species-specific prior knowledge of microsatellite location or composition. Very small amounts of DNA are required, making this method ideal for organisms of conservation concern, or where the quantity of DNA is extremely limited due to organism size. ISSR-PCR can be highly reproducible but requires careful attention to detail. Optimization of DNA extraction, fragment amplification, and normalization of fragment peak heights during fluorescent detection are critical steps to minimizing the downstream time spent verifying and scoring the data.

Contagious Deposition of Seeds in Spider Monkeys' Sleeping Trees Limits Effective Seed Dispersal in Fragmented Landscapes

PubMed Central

González-Zamora, Arturo; Arroyo-Rodríguez, Víctor; Escobar, Federico; Rös, Matthias; Oyama, Ken; Ibarra-Manríquez, Guillermo; Stoner, Kathryn E.; Chapman, Colin A.

2014-01-01

The repeated use of sleeping sites by frugivorous vertebrates promotes the deposition and aggregation of copious amounts of seeds in these sites. This spatially contagious pattern of seed deposition has key implications for seed dispersal, particularly because such patterns can persist through recruitment. Assessing the seed rain patterns in sleeping sites thus represents a fundamental step in understanding the spatial structure and regeneration of plant assemblages. We evaluated the seed rain produced by spider monkeys (Ateles geoffroyi) in latrines located beneath 60 sleeping trees in two continuous forest sites (CFS) and three forest fragments (FF) in the Lacandona rainforest, Mexico. We tested for differences among latrines, among sites, and between forest conditions in the abundance, diversity (α-, β- and, γ-components) and evenness of seed assemblages. We recorded 45,919 seeds ≥5 mm (in length) from 68 species. The abundance of seeds was 1.7 times higher in FF than in CFS, particularly because of the dominance of a few plant species. As a consequence, community evenness tended to be lower within FF. β-diversity of common and dominant species was two times greater among FF than between CFS. Although mean α-diversity per latrine did not differ among sites, the greater β-diversity among latrines in CFS increased γ-diversity in these sites, particularly when considering common and dominant species. Our results support the hypothesis that fruit scarcity in FF can ‘force’ spider monkeys to deplete the available fruit patches more intensively than in CFS. This feeding strategy can limit the effectiveness of spider monkeys as seed dispersers in FF, because (i) it can limit the number of seed dispersers visiting such fruit patches; (ii) it increases seed dispersal limitation; and (iii) it can contribute to the floristic homogenization (i.e., reduced β-diversity among latrines) in fragmented landscapes. PMID:24586705

Continuing Education in Postmodernity: Four Semantic Tensions.

ERIC Educational Resources Information Center

Bagnall, Richard G.

1994-01-01

Describes four tensions in postmodern continuing education practice: aggregation versus fragmentation, consistency versus flexibility, description versus evaluation, and associationality versus immediacy. Explains the two opposing inclinations in these concepts: there are good reasons following each one, but doing so limits the possibility of…

Traces of pFc' in IVIG interact with human IgG Fc domains and counteract aggregation.

PubMed

Rispens, Theo; Himly, Martin; Ooievaar-De Heer, Pleuni; den Bleker, Tamara H; Aalberse, Rob C

2010-04-16

To prevent multimer formation, intravenous immunoglobulin (IVIG) is often treated with traces of pepsin. So far, the mechanism behind this treatment has been unclear. Recently, we reported that human IgG4 binds other IgG molecules via Fc-Fc interactions. Here we show that IVIG treated with traces of pepsin (Nanogam) inhibits these interactions. We found that--besides IgG4--peptides corresponding to IgG1 and IgG2 pFc' (products of limited pepsin digestion) are responsible for the inhibitory action. Using radiolabeled pFc', it was found that pFc' binds directly to IgG1. Furthermore, recombinant CH3 fragments were found to also possess binding activity, and potencies of inhibition varied over 3 orders of magnitude amongst the subclasses, IgG4 being most potent. We propose that pFc' formation explains how limited pepsin digestion diminishes adverse effects of IVIG. In particular, the presence of this fragment can enhance the stability of IgG products including IVIG and therapeutical monoclonal antibodies. Indeed, using a model system it was found that acid-induced aggregation of IgG is reduced in the presence of pFc', suggesting a 'chaperone-like' activity of this fragment. Thus, pFc' can modulate Fc interactions and may therefore reduce adverse effects of IVIG, in particular by preventing oligomerization. 2010 Elsevier B.V. All rights reserved.

Adaptations in Electronic Structure Calculations in Heterogeneous Environments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Talamudupula, Sai

Modern quantum chemistry deals with electronic structure calculations of unprecedented complexity and accuracy. They demand full power of high-performance computing and must be in tune with the given architecture for superior e ciency. To make such applications resourceaware, it is desirable to enable their static and dynamic adaptations using some external software (middleware), which may monitor both system availability and application needs, rather than mix science with system-related calls inside the application. The present work investigates scienti c application interlinking with middleware based on the example of the computational chemistry package GAMESS and middleware NICAN. The existing synchronous model ismore » limited by the possible delays due to the middleware processing time under the sustainable runtime system conditions. Proposed asynchronous and hybrid models aim at overcoming this limitation. When linked with NICAN, the fragment molecular orbital (FMO) method is capable of adapting statically and dynamically its fragment scheduling policy based on the computing platform conditions. Signi cant execution time and throughput gains have been obtained due to such static adaptations when the compute nodes have very di erent core counts. Dynamic adaptations are based on the main memory availability at run time. NICAN prompts FMO to postpone scheduling certain fragments, if there is not enough memory for their immediate execution. Hence, FMO may be able to complete the calculations whereas without such adaptations it aborts.« less

Interaction energies for the purine inhibitor roscovitine with cyclin-dependent kinase 2: correlated ab initio quantum-chemical, DFT and empirical calculations.

PubMed

Dobes, Petr; Otyepka, Michal; Strnad, Miroslav; Hobza, Pavel

2006-05-24

The interaction between roscovitine and cyclin-dependent kinase 2 (cdk2) was investigated by performing correlated ab initio quantum-chemical calculations. The whole protein was fragmented into smaller systems consisting of one or a few amino acids, and the interaction energies of these fragments with roscovitine were determined by using the MP2 method with the extended aug-cc-pVDZ basis set. For selected complexes, the complete basis set limit MP2 interaction energies, as well as the coupled-cluster corrections with inclusion of single, double and noninteractive triples contributions [CCSD(T)], were also evaluated. The energies of interaction between roscovitine and small fragments and between roscovitine and substantial sections of protein (722 atoms) were also computed by using density-functional tight-binding methods covering dispersion energy (DFTB-D) and the Cornell empirical potential. Total stabilisation energy originates predominantly from dispersion energy and methods that do not account for the dispersion energy cannot, therefore, be recommended for the study of protein-inhibitor interactions. The Cornell empirical potential describes reasonably well the interaction between roscovitine and protein; therefore, this method can be applied in future thermodynamic calculations. A limited number of amino acid residues contribute significantly to the binding of roscovitine and cdk2, whereas a rather large number of amino acids make a negligible contribution.

Detection of two fungal biocontrol agents against root-knot nematodes by RAPD markers.

PubMed

Zhu, Ming Liang; Mo, Ming He; Xia, Zhen Yuan; Li, Yun Hua; Yang, Shu Jun; Li, Tian Fei; Zhang, Ke Qin

2006-05-01

The strain ZK7 of Pochonia chlamydosporia var. chlamydosporia and IPC of Paecilomyces lilacinus are highly effective in the biological control against root-knot nematodes infecting tobacco. When applied, they require a specific monitoring method to evaluate the colonization and dispersal in soil. In this work, the randomly amplified polymorphic DNA (RAPD) technique was used to differentiate between the two individual strains and 95 other isolates, including isolates of the same species and common soil fungi. This approach allowed the selection of specific fragments of 1.2 kb (Vc1200) and 2.0 kb (Vc2000) specific for ZK7, 1.4 kb (P1400) and 0.85 kb (P850) specific for IPC, using the random Primers OPL-02, OPD-05, OPD-05 and OPC-11, respectively. These fragments were cloned, sequenced, and used to design sequence-characterized amplification region (SCAR) primers specific for the two strains. In classical polymerase chain reaction (PCR), with serial dilution of ZK7 and IPC pure culture DNAs template, the detection limits of these oligonucleotide SCAR-PCR primers were found to be 10, 1000, 500, 100 pg, respectively. In the dot blotting, digoxigenin (DIG)-labeled amplicons from these four primers specifically recognized the corresponding fragments in the DNAs template of these two strains. The detection limit of these amplicons were 0.2, 0.2, 0.5, 0.5 mug, respectively.

Imaging Systems for Size Measurements of Debrisat Fragments

NASA Technical Reports Server (NTRS)

Shiotani, B.; Scruggs, T.; Toledo, R.; Fitz-Coy, N.; Liou, J. C.; Sorge, M.; Huynh, T.; Opiela, J.; Krisko, P.; Cowardin, H.

2017-01-01

The overall objective of the DebriSat project is to provide data to update existing standard spacecraft breakup models. One of the key sets of parameters used in these models is the physical dimensions of the fragments (i.e., length, average-cross sectional area, and volume). For the DebriSat project, only fragments with at least one dimension greater than 2 mm are collected and processed. Additionally, a significant portion of the fragments recovered from the impact test are needle-like and/or flat plate-like fragments where their heights are almost negligible in comparison to their other dimensions. As a result, two fragment size categories were defined: 2D objects and 3D objects. While measurement systems are commercially available, factors such as measurement rates, system adaptability, size characterization limitations and equipment costs presented significant challenges to the project and a decision was made to develop our own size characterization systems. The size characterization systems consist of two automated image systems, one referred to as the 3D imaging system and the other as the 2D imaging system. Which imaging system to use depends on the classification of the fragment being measured. Both imaging systems utilize point-and-shoot cameras for object image acquisition and create representative point clouds of the fragments. The 3D imaging system utilizes a space-carving algorithm to generate a 3D point cloud, while the 2D imaging system utilizes an edge detection algorithm to generate a 2D point cloud. From the point clouds, the three largest orthogonal dimensions are determined using a convex hull algorithm. For 3D objects, in addition to the three largest orthogonal dimensions, the volume is computed via an alpha-shape algorithm applied to the point clouds. The average cross-sectional area is also computed for 3D objects. Both imaging systems have automated size measurements (image acquisition and image processing) driven by the need to quickly and accurately measure tens of thousands of debris fragments. Moreover, the automated size measurement reduces potential fragment damage/mishandling and ability for accuracy and repeatability. As the fragment characterization progressed, it became evident that the imaging systems had to be revised. For example, an additional view was added to the 2D imaging system to capture the height of the 2D object. This paper presents the DebriSat project's imaging systems and calculation techniques in detail; from design and development to maturation. The experiences and challenges are also shared.

Design of character-based DNA barcode motif for species identification: A computational approach and its validation in fishes.

PubMed

Chakraborty, Mohua; Dhar, Bishal; Ghosh, Sankar Kumar

2017-11-01

The DNA barcodes are generally interpreted using distance-based and character-based methods. The former uses clustering of comparable groups, based on the relative genetic distance, while the latter is based on the presence or absence of discrete nucleotide substitutions. The distance-based approach has a limitation in defining a universal species boundary across the taxa as the rate of mtDNA evolution is not constant throughout the taxa. However, character-based approach more accurately defines this using a unique set of nucleotide characters. The character-based analysis of full-length barcode has some inherent limitations, like sequencing of the full-length barcode, use of a sparse-data matrix and lack of a uniform diagnostic position for each group. A short continuous stretch of a fragment can be used to resolve the limitations. Here, we observe that a 154-bp fragment, from the transversion-rich domain of 1367 COI barcode sequences can successfully delimit species in the three most diverse orders of freshwater fishes. This fragment is used to design species-specific barcode motifs for 109 species by the character-based method, which successfully identifies the correct species using a pattern-matching program. The motifs also correctly identify geographically isolated population of the Cypriniformes species. Further, this region is validated as a species-specific mini-barcode for freshwater fishes by successful PCR amplification and sequencing of the motif (154 bp) using the designed primers. We anticipate that use of such motifs will enhance the diagnostic power of DNA barcode, and the mini-barcode approach will greatly benefit the field-based system of rapid species identification. © 2017 John Wiley & Sons Ltd.

Overcoming codon-usage bias in heterologous protein expression in Streptococcus gordonii.

PubMed

Lee, Song F; Li, Yi-Jing; Halperin, Scott A

2009-11-01

One of the limitations facing the development of Streptococcus gordonii into a successful vaccine vector is the inability of this bacterium to express high levels of heterologous proteins. In the present study, we have identified 12 codons deemed as rare codons in S. gordonii and seven other streptococcal species. tRNA genes encoding 10 of the 12 rare codons were cloned into a plasmid. The plasmid was transformed into strains of S. gordonii expressing the fusion protein SpaP/S1, the anti-complement receptor 1 (CR1) single-chain variable fragment (scFv) antibody, or the Toxoplasma gondii cyclophilin C18 protein. These three heterologous proteins contained high percentages of amino acids encoded by rare codons. The results showed that the production of SpaP/S1, anti-CR1 scFv and C18 increased by 2.7-, 120- and 10-fold, respectively, over the control strains. In contrast, the production of the streptococcal SpaP protein without the pertussis toxin S1 fragment was not affected by tRNA gene supplementation, indicating that the increased production of SpaP/S1 protein was due to the ability to overcome the limitation caused by rare codons required for the S1 fragment. The increase in anti-CR1 scFv production was also observed in Streptococcus mutans following tRNA gene supplementation. Collectively, the findings in the present study demonstrate for the first time, to the best of our knowledge, that codon-usage bias exists in Streptococcus spp. and the limitation of heterologous protein expression caused by codon-usage bias can be overcome by tRNA supplementation.

Impact of MR-guided boiling histotripsy in distinct murine tumor models.

PubMed

Hoogenboom, Martijn; Eikelenboom, Dylan C; van den Bijgaart, Renske J E; Heerschap, Arend; Wesseling, Pieter; den Brok, Martijn H; Fütterer, Jurgen J; Adema, Gosse J

2017-09-01

Interest in mechanical high intensity focused ultrasound (HIFU) ablation is rapidly growing. Boiling histotripsy (BH) is applied for mechanical fragmentation of soft tissue into submicron fragments with limited temperature increase using the shock wave and cavitation effects of HIFU. Research on BH has been largely limited to ex vivo experiments. As a consequence, the in vivo pathology after BH treatment and the relation to preexistent tissue characteristics are not well understood. This study reports on in vivo MR guided BH treatment, either with 100 or 200 pulses per focal spot, in three different subcutaneous mouse tumor models: a soft-tissue melanoma (B16OVA), a compact growing thymoma (EL4), and a highly vascularized neuroblastoma (9464D). Extensive treatment evaluation was performed using MR imaging followed by histopathology 2h after treatment. T2 weighted MRI allowed direct in vivo visualization of the BH lesions in all tumor models. The 100-pulse treated area in the B16OVA tumors was larger than the predicted treatment volume (500±10%). For the more compact growing EL4 and 9464D tumors this was 95±13% and 55±33%, respectively. Histopathology after the 100-pulse treatment revealed completely disintegrated lesions in the treated area with sharp borders in the compact EL4 and 9464D tumors, while for B16OVA tumors the lesion contained a mixture of discohesive (partly viable) clusters of cells, micro-vessel remainings, and tumor cell debris. The treatment of B16OVA with 200 pulses increased the fragmentation of tumor tissue. In all tumor types only micro-hemorrhages were detected after ablation (slightly higher after 200-pulse treatment for the highly vascularized 9464D tumors). Collagen staining revealed that the collagen fibers were to a greater or lesser extent still intact and partly clotted together near the lesion border in all tumor models. In conclusion, this study reveals effective mechanical fragmentation of different tumor types using BH without major hemorrhages. However, treatment settings may need to be adjusted to the tissue characteristics for optimal tissue fragmentation. Copyright © 2017 Elsevier B.V. All rights reserved.

Hybridization Capture Using RAD Probes (hyRAD), a New Tool for Performing Genomic Analyses on Collection Specimens

PubMed Central

Suchan, Tomasz; Pitteloud, Camille; Gerasimova, Nadezhda S.; Kostikova, Anna; Schmid, Sarah; Arrigo, Nils; Pajkovic, Mila; Ronikier, Michał; Alvarez, Nadir

2016-01-01

In the recent years, many protocols aimed at reproducibly sequencing reduced-genome subsets in non-model organisms have been published. Among them, RAD-sequencing is one of the most widely used. It relies on digesting DNA with specific restriction enzymes and performing size selection on the resulting fragments. Despite its acknowledged utility, this method is of limited use with degraded DNA samples, such as those isolated from museum specimens, as these samples are less likely to harbor fragments long enough to comprise two restriction sites making possible ligation of the adapter sequences (in the case of double-digest RAD) or performing size selection of the resulting fragments (in the case of single-digest RAD). Here, we address these limitations by presenting a novel method called hybridization RAD (hyRAD). In this approach, biotinylated RAD fragments, covering a random fraction of the genome, are used as baits for capturing homologous fragments from genomic shotgun sequencing libraries. This simple and cost-effective approach allows sequencing of orthologous loci even from highly degraded DNA samples, opening new avenues of research in the field of museum genomics. Not relying on the restriction site presence, it improves among-sample loci coverage. In a trial study, hyRAD allowed us to obtain a large set of orthologous loci from fresh and museum samples from a non-model butterfly species, with a high proportion of single nucleotide polymorphisms present in all eight analyzed specimens, including 58-year-old museum samples. The utility of the method was further validated using 49 museum and fresh samples of a Palearctic grasshopper species for which the spatial genetic structure was previously assessed using mtDNA amplicons. The application of the method is eventually discussed in a wider context. As it does not rely on the restriction site presence, it is therefore not sensitive to among-sample loci polymorphisms in the restriction sites that usually causes loci dropout. This should enable the application of hyRAD to analyses at broader evolutionary scales. PMID:26999359

Rapid genetic restoration of a keystone species exhibiting delayed demographic response

USDA-ARS?s Scientific Manuscript database

Genetic founder effects are often expected when animals colonize restored habitat in fragmented landscapes, but empirical data on genetic responses to restoration are limited. We examined the genetic response of banner-tailed kangaroo rats (Dipodomys spectabilis) to landscape-scale grassland restor...

Vulnerability of Northeastern U.S. Salt Marshes to Climatic and Anthropogenic Stressors

EPA Science Inventory

In the Northeastern U.S., salt marsh area is in decline. Habitat change analysis has revealed fragmentation, displacement of high marsh by low marsh species, and marsh drowning, while development of adjacent uplands limits upslope migration. Using inundation experiments, field s...

Vulnerability of Northeastern U.S. Salt Marshes to Climatic and Anthropogenic Stressors (AGU)

EPA Science Inventory

In the Northeastern U.S., salt marsh area is in decline. Habitat change analysis has revealed fragmentation, displacement of high marsh by low marsh species, and ecological drowning, while development of adjacent uplands limits upslope migration. Using inundation experiments, fi...

Xylan hydrolysis in Populus trichocarpa × P. deltoides and model substrates during hydrothermal pretreatment.

PubMed

Trajano, Heather L; Pattathil, Sivakumar; Tomkins, Bruce A; Tschaplinski, Timothy J; Hahn, Michael G; Van Berkel, Gary J; Wyman, Charles E

2015-03-01

Previous studies defined easy and difficult to hydrolyze fractions of hemicellulose that may result from bonds among cellulose, hemicellulose, and lignin. To understand how such bonds affect hydrolysis, Populus trichocarpa × Populus deltoides, holocellulose isolated from P. trichocarpa × P. deltoides and birchwood xylan were subjected to hydrothermal flow-through pretreatment. Samples were characterized by glycome profiling, HPLC, and UPLC-MS. Glycome profiling revealed steady fragmentation and removal of glycans from solids during hydrolysis. The extent of polysaccharide fragmentation, hydrolysis rate, and total xylose yield were lowest for P. trichocarpa × P. deltoides and greatest for birchwood xylan. Comparison of results from P. trichocarpa × P. deltoides and holocellulose suggested that lignin-carbohydrate complexes reduce hydrolysis rates and limit release of large xylooligomers. Smaller differences between results with holocellulose and birchwood xylan suggest xylan-cellulose hydrogen bonds limited hydrolysis, but to a lesser extent. These findings imply cell wall structure strongly influences hydrolysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

Mass Spectrometry Based Ultrasensitive DNA Methylation Profiling Using Target Fragmentation Assay.

PubMed

Lin, Xiang-Cheng; Zhang, Ting; Liu, Lan; Tang, Hao; Yu, Ru-Qin; Jiang, Jian-Hui

2016-01-19

Efficient tools for profiling DNA methylation in specific genes are essential for epigenetics and clinical diagnostics. Current DNA methylation profiling techniques have been limited by inconvenient implementation, requirements of specific reagents, and inferior accuracy in quantifying methylation degree. We develop a novel mass spectrometry method, target fragmentation assay (TFA), which enable to profile methylation in specific sequences. This method combines selective capture of DNA target from restricted cleavage of genomic DNA using magnetic separation with MS detection of the nonenzymatic hydrolysates of target DNA. This method is shown to be highly sensitive with a detection limit as low as 0.056 amol, allowing direct profiling of methylation using genome DNA without preamplification. Moreover, this method offers a unique advantage in accurately determining DNA methylation level. The clinical applicability was demonstrated by DNA methylation analysis using prostate tissue samples, implying the potential of this method as a useful tool for DNA methylation profiling in early detection of related diseases.

Are habitat fragmentation, local adaptation and isolation-by-distance driving population divergence in wild rice Oryza rufipogon?

PubMed

Zhao, Yao; Vrieling, Klaas; Liao, Hui; Xiao, Manqiu; Zhu, Yongqing; Rong, Jun; Zhang, Wenju; Wang, Yuguo; Yang, Ji; Chen, Jiakuan; Song, Zhiping

2013-11-01

Habitat fragmentation weakens the connection between populations and is accompanied with isolation by distance (IBD) and local adaptation (isolation by adaptation, IBA), both leading to genetic divergence between populations. To understand the evolutionary potential of a population and to formulate proper conservation strategies, information on the roles of IBD and IBA in driving population divergence is critical. The putative ancestor of Asian cultivated rice (Oryza sativa) is endangered in China due to habitat loss and fragmentation. We investigated the genetic variation in 11 Chinese Oryza rufipogon populations using 79 microsatellite loci to infer the effects of habitat fragmentation, IBD and IBA on genetic structure. Historical and current gene flows were found to be rare (mh  = 0.0002-0.0013, mc  = 0.007-0.029), indicating IBD and resulting in a high level of population divergence (FST  = 0.343). High within-population genetic variation (HE  = 0.377-0.515), relatively large effective population sizes (Ne  = 96-158), absence of bottlenecks and limited gene flow were found, demonstrating little impact of recent habitat fragmentation on these populations. Eleven gene-linked microsatellite loci were identified as outliers, indicating local adaptation. Hierarchical AMOVA and partial Mantel tests indicated that population divergence of Chinese O. rufipogon was significantly correlated with environmental factors, especially habitat temperature. Common garden trials detected a significant adaptive population divergence associated with latitude. Collectively, these findings imply that IBD due to historical rather than recent fragmentation, followed by local adaptation, has driven population divergence in O. rufipogon. © 2013 John Wiley & Sons Ltd.

Conserved stem fragment from H3 influenza hemagglutinin elicits cross-clade neutralizing antibodies through stalk-targeted blocking of conformational change during membrane fusion.

PubMed

Gong, Xin; Yin, He; Shi, Yuhua; Guan, Shanshan; He, Xiaoqiu; Yang, Lan; Yu, Yongjiao; Kuai, Ziyu; Jiang, Chunlai; Kong, Wei; Wang, Song; Shan, Yaming

2016-04-01

Currently available influenza vaccines typically fail to elicit/boost broadly neutralizing antibodies due to the mutability of virus sequences and conformational changes during protective immunity, thereby limiting their efficacy. This problem needs to be addressed by further understanding the mechanisms of neutralization and finding the desired neutralizing site during membrane fusion. This study specifically focused on viruses of the H3N2 subtype, which have persisted as a principal source of influenza-related morbidity and mortality in humans since the 1968 influenza pandemic. Through sequence alignment and epitope prediction, a series of highly conserved stem fragments (spanning 47 years) were found and coupled to the Keyhole Limpet Hemocyanin (KLH) protein. By application of a combinatorial display library and crystal structure modeling, a stem fragment immunogen, located at the turning point of the HA neck undergoing conformational change during membrane fusion with both B- and T-cell epitopes, was identified. After synthesis of the optimal stem fragment using a multiple antigen peptide (MAP) system, strong humoral immune responses and cross-clade neutralizing activities against strains from the H3 subtype of group 2 influenza viruses after animal immunizations were observed. By detection of nuclear protein immunofluorescence with acid bypass treatment, antisera raised against MAP4 immunogens of the stem fragment showed the potential to inhibit the conformational change of HA in stem-targeted virus neutralization. The identification of this conserved stem fragment provides great potential for exploitation of this site of vulnerability in therapeutic and vaccine design. Copyright © 2016 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.

Does landscape fragmentation influence sex ratio of dioecious plants? A case study of Pistacia chinensis in the Thousand-Island Lake region of China.

PubMed

Yu, Lin; Lu, Jianbo

2011-01-01

The Thousand-Island Lake region in Zhejiang Province, China is a highly fragmented landscape with a clear point-in-time of fragmentation as a result of flooding to form the reservoir. Islands in the artificial lake were surveyed to examine how population sex ratio of a dioecious plant specie Pistacia chinensis B. was affected by landscape fragmentation. A natural population on the mainland near the lake was also surveyed for comparison. Population size, sex ratio and diameter at breast height (DBH) of individuals were measured over 2 years. More than 1,500 individuals, distributed in 31 populations, were studied. Soil nitrogen in the different populations was measured to identify the relationship between sex ratio and micro-environmental conditions. In accordance with the results of many other reports on biased sex ratio in relation to environmental gradient, we found that poor soil nitrogen areas fostered male-biased populations. In addition, the degree of sex ratio bias increased with decreasing population size and population connectivity. The biased sex ratios were only found in younger individuals (less than 50 years old) in small populations, while a stable 1∶1 sex ratio was found in the large population on the mainland. We concluded that the effects of landscape fragmentation on the dioecious population sex ratio were mainly achieved in relation to changing soil nitrogen conditions in patches and pollen limitation within and among populations. Large populations could maintain a more suitable environment in terms of nutrient conditions and pollen flow, subsequently maintaining a stable sex ratio in dioecious plant populations. Both micro-environmental factors and spatial structure should be considered in fragmented landscape for the conservation of dioecious plant species.

Macromolecular ab initio phasing enforcing secondary and tertiary structure.

PubMed

Millán, Claudia; Sammito, Massimo; Usón, Isabel

2015-01-01

Ab initio phasing of macromolecular structures, from the native intensities alone with no experimental phase information or previous particular structural knowledge, has been the object of a long quest, limited by two main barriers: structure size and resolution of the data. Current approaches to extend the scope of ab initio phasing include use of the Patterson function, density modification and data extrapolation. The authors' approach relies on the combination of locating model fragments such as polyalanine α-helices with the program PHASER and density modification with the program SHELXE. Given the difficulties in discriminating correct small substructures, many putative groups of fragments have to be tested in parallel; thus calculations are performed in a grid or supercomputer. The method has been named after the Italian painter Arcimboldo, who used to compose portraits out of fruit and vegetables. With ARCIMBOLDO, most collections of fragments remain a 'still-life', but some are correct enough for density modification and main-chain tracing to reveal the protein's true portrait. Beyond α-helices, other fragments can be exploited in an analogous way: libraries of helices with modelled side chains, β-strands, predictable fragments such as DNA-binding folds or fragments selected from distant homologues up to libraries of small local folds that are used to enforce nonspecific tertiary structure; thus restoring the ab initio nature of the method. Using these methods, a number of unknown macromolecules with a few thousand atoms and resolutions around 2 Å have been solved. In the 2014 release, use of the program has been simplified. The software mediates the use of massive computing to automate the grid access required in difficult cases but may also run on a single multicore workstation (http://chango.ibmb.csic.es/ARCIMBOLDO_LITE) to solve straightforward cases.

Comparison of Drosophilidae (Diptera) assemblages from two highland Araucaria Forest fragments, with and without environmental conservation policies.

PubMed

Cavasini, R; Buschini, M L T; Machado, L P B; Mateus, R P

2014-11-01

Flies from the Drosophilidae family are model organisms for biological studies and are often suggested as bioindicators of environmental quality. The Araucaria Forest, one of Atlantic Forest phyto-physiognomy, displays a highly fragmented distribution due to the expansion of agriculture and urbanization. Thus, this work aimed to evaluate and compare the drosophilid assemblages from two highland Araucaria Forest fragments, one a conservation unit (PMA - Parque Municipal das Araucárias) and the other a private property without any conservational policy (FBL - Fazenda Brandalise), in space and time, using species abundances and richness, ecological indexes and Neotropical and exotic species proportions as parameters to establish the level of environmental quality of these fragments. Our results showed that the observed diversity in PMA (H' = 2.221) was approximately 40% higher than in FBL (H' = 1.592). This could be due to higher preservation quality and habitat diversity in PMA, indicating the importance of conservation units. However, richness were similar for these areas, with PMA (Dmg = 6.602) only 8% higher than FBL (Dmg = 6.128), which suggest that the larger distance from city limits and the larger size of FBL forested area could be compensating the higher disturbance caused by antrophic extractive exploitation of this fragment. This points out that, besides the quality of presevertion, the size and/or connection with other fragments should be considered for areas destined for biodiversity conservation. In general, both areas presented similar drosophilid assemblages, and the expressive abundance of both Neotropical species (mostly of the subgroup willistoni) and the exotic species D. kikkawai suggests that these areas are in intermediate stages of conservation.

Does the seed bank contribute to the build-up of a genetic extinction debt in the grassland perennial Campanula rotundifolia?

PubMed

Plue, Jan; Vandepitte, Katrien; Honnay, Olivier; Cousins, Sara A O

2017-09-01

Habitat fragmentation threatens global biodiversity. Many plant species persist in habitat fragments via persistent life cycle stages such as seed banks, generating a species extinction debt. Here, seed banks are hypothesized to cause a temporal delay in the expected loss of genetic variation, which can be referred to as a genetic extinction debt, as a possible mechanism behind species extinction debts. Fragmented grassland populations of Campanula rotundifolia were examined for evidence of a genetic extinction debt, investigating if the seed bank contributed to the extinction debt build-up. The genetic make-up of 15 above- and below-ground populations was analysed in relation to historical and current levels of habitat fragmentation, both separately and combined. Genetic diversity was highest in above-ground populations, though below-ground populations contained 8 % of unique alleles that were absent above-ground. Above-ground genetic diversity and composition were related to historical patch size and connectivity, but not current patch characteristics, suggesting the presence of a genetic extinction debt in the above-ground populations. No such relationships were found for the below-ground populations. Genetic diversity measures still showed a response to historical but not present landscape characteristics when combining genetic diversity of the above- and below-ground populations. The fragmented C. rotundifolia populations exhibited a genetic extinction debt. However, the role of the seed banks in the build-up of this extinction debt is probably small, since the limited, unique genetic diversity of the seed bank alone seems unable to counter the detrimental effects of habitat fragmentation on the population genetic structure of C. rotundifolia. © The Author 2017. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Consequences of Landscape Fragmentation on Lyme Disease Risk: A Cellular Automata Approach

PubMed Central

Li, Sen; Hartemink, Nienke; Speybroeck, Niko; Vanwambeke, Sophie O.

2012-01-01

The abundance of infected Ixodid ticks is an important component of human risk of Lyme disease, and various empirical studies have shown that this is associated, at least in part, to landscape fragmentation. In this study, we aimed at exploring how varying woodland fragmentation patterns affect the risk of Lyme disease, through infected tick abundance. A cellular automata model was developed, incorporating a heterogeneous landscape with three interactive components: an age-structured tick population, a classical disease transmission function, and hosts. A set of simplifying assumptions were adopted with respect to the study objective and field data limitations. In the model, the landscape influences both tick survival and host movement. The validation of the model was performed with an empirical study. Scenarios of various landscape configurations (focusing on woodland fragmentation) were simulated and compared. Lyme disease risk indices (density and infection prevalence of nymphs) differed considerably between scenarios: (i) the risk could be higher in highly fragmented woodlands, which is supported by a number of recently published empirical studies, and (ii) grassland could reduce the risk in adjacent woodland, which suggests landscape fragmentation studies of zoonotic diseases should not focus on the patch-level woodland patterns only, but also on landscape-level adjacent land cover patterns. Further analysis of the simulation results indicated strong correlations between Lyme disease risk indices and the density, shape and aggregation level of woodland patches. These findings highlight the strong effect of the spatial patterns of local host population and movement on the spatial dynamics of Lyme disease risks, which can be shaped by woodland fragmentation. In conclusion, using a cellular automata approach is beneficial for modelling complex zoonotic transmission systems as it can be combined with either real world landscapes for exploring direct spatial effects or artificial representations for outlining possible empirical investigations. PMID:22761842

A comparison of capture antibody fragments in cardiac troponin I immunoassay.

PubMed

Hyytiä, Heidi; Järvenpää, Marja-Leena; Ristiniemi, Noora; Lövgren, Timo; Pettersson, Kim

2013-08-01

To compare cardiac troponin I (cTnI) values measured from 32 normal plasma specimens with a two-site cTnI research assay exploiting different molecular forms of a capture antibody. The current research assay consists of two capture antibodies immobilized on streptavidin-well surface and one detection antibody attached to highly fluorescent europium(III)-chelate-doped nanoparticles. Four different molecular forms of one of the capture antibodies (intact monoclonal (Mab), F(ab')2 fragment, Fab fragment and chimeric Fab fragment (cFab)) were tested. The developed immunoassays were evaluated in terms of their analytical sensitivities and assay kinetics. Furthermore, cTnI concentrations were measured from 32 heparin plasma samples from apparently healthy donors (mean age 32; range 24-60 years). The differences in the measured cTnI concentrations (corrected for the buffer-based zero calibrator) between the Mab and the three fragmented forms were highly significant (P<0.0001). Replacing the intact Mab with the antibody fragments also reduced the required antibody amount from 100 ng to 66 ng (F(ab')2) and 16.5 ng (Fab and cFab). Furthermore, the limit of detection was improved when Fab fragments were employed (Mab: 0.90 ng/L, Fab: 0.69 ng/L and cFab: 0.41 ng/L). The apparent normal range median (minimum/maximum) of the 32 healthy subjects was reduced from 7.28 ng/L (2.64/116 ng/L) with Mab to 1.80 ng/L (0.746/10.6 ng/L) for the cFab. Eliminating the Fc-part from one of the two capture antibodies in an immunofluorometric cTnI assay substantially reduced the measured cTnI concentrations, simultaneously improving the assay sensitivity and reducing the reagent consumption. Copyright © 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Establishment of hapten-specific monoclonal avian IgY by conversion of antibody fragments obtained from combinatorial libraries.

PubMed

Deckers, Susanne; Braren, Ingke; Greunke, Kerstin; Meyer, Nadine; Rühl, Dana; Bredehorst, Reinhard; Spillner, Edzard

2009-01-01

Nowadays, recombinant antibody and phage display technology enable the efficient generation of immunotools and a subsequent manipulation for optimized affinity, specificity or overall performance. Such advantages are of particular interest for haptenic target structures, such as TNT (2,4,6-trinitrotoluene). The toxicity of TNT and its breakdown products makes a reliable and fast detection of low levels in aqueous samples highly important. In the present study, we aimed for the generation of scFvs (single-chain antibody fragments) specific for the TNT-surrogate TNP (2,4,6-trinitrophenyl) and their subsequent production as monoclonal avian IgY immunoglobulins providing improved assay performance. Therefore we subjected a human synthetic scFv library to selection following different strategies. TNP-specific human antibody fragments could be identified, characterized for their primary structure and evaluated for production as soluble scFv in Escherichia coli. Additionally, a murine TNP-specific antibody fragment was obtained from the hybridoma 11B3; however, the prokaryotic expression level was found to be limited. To generate and evaluate immunoglobulin formats with superior characteristics, all recombinant antibody fragments then were converted into two different chimaeric bivalent IgY antibody formats. After expression in mammalian cells, the IgY antibodies were assessed for their reactivity towards TNT. The IgY antibodies generated on the basis of the combinatorial library proved to be useful for detection of TNT, thereby emphasizing the high potential of this approach for the development of detection devices for immunoassay-based techniques.

Theoretical Investigations of Well-Defined Graphene Nanostructures: Catalysis, Spectroscopy, and Development of Novel Fragment-Based Models

NASA Astrophysics Data System (ADS)

Noffke, Benjamin W.

Carbon materials have the potential to replace some precious metals in renewable energy applications. These materials are particularly attractive because of the elemental abundance and relatively low nuclear mass of carbon, implying economically feasible and lightweight materials. Targeted design of carbon materials is hindered by the lack of fundamental understanding that is required to tailor their properties for the desired application. However, most available synthetic methods to create carbon materials involve harsh conditions that limit the control of the resulting structure. Without a well-defined structure, the system is too complex and fundamental studies cannot be definitive. This work seeks to gain fundamental understanding through the development and application of efficient computational models for these systems, in conjunction with experiments performed on soluble, well-defined graphene nanostructures prepared by our group using a bottom-up synthetic approach. Theory is used to determine mechanistic details for well-defined carbon systems in applications of catalysis and electrochemical transformations. The resulting computational models do well to explain previous observations of carbon materials and provide suggestions for future directions. However, as the system size of the nanostructures gets larger, the computational cost can become prohibitive. To reduce the computational scaling of quantum chemical calculations, a new fragmentation scheme has been developed that addresses the challenges of fragmenting conjugated molecules. By selecting fragments that retain important structural characteristics in graphene, a more efficient method is achieved. The new method paves the way for an automated, systematic fragmentation scheme of graphene molecules.

SKI2 mediates degradation of RISC 5'-cleavage fragments and prevents secondary siRNA production from miRNA targets in Arabidopsis.

PubMed

Branscheid, Anja; Marchais, Antonin; Schott, Gregory; Lange, Heike; Gagliardi, Dominique; Andersen, Stig Uggerhøj; Voinnet, Olivier; Brodersen, Peter

2015-12-15

Small regulatory RNAs are fundamental in eukaryotic and prokaryotic gene regulation. In plants, an important element of post-transcriptional control is effected by 20-24 nt microRNAs (miRNAs) and short interfering RNAs (siRNAs) bound to the ARGONAUTE1 (AGO1) protein in an RNA induced silencing complex (RISC). AGO1 may cleave target mRNAs with small RNA complementarity, but the fate of the resulting cleavage fragments remains incompletely understood. Here, we show that SKI2, SKI3 and SKI8, subunits of a cytoplasmic cofactor of the RNA exosome, are required for degradation of RISC 5', but not 3'-cleavage fragments in Arabidopsis. In the absence of SKI2 activity, many miRNA targets produce siRNAs via the RNA-dependent RNA polymerase 6 (RDR6) pathway. These siRNAs are low-abundant, and map close to the cleavage site. In most cases, siRNAs were produced 5' to the cleavage site, but several examples of 3'-spreading were also identified. These observations suggest that siRNAs do not simply derive from RDR6 action on stable 5'-cleavage fragments and hence that SKI2 has a direct role in limiting secondary siRNA production in addition to its function in mediating degradation of 5'-cleavage fragments. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Evidence for Sequence Scrambling and Divergent H/D Exchange Reactions of Doubly-Charged Isobaric b-Type Fragment Ions

NASA Astrophysics Data System (ADS)

Zekavat, Behrooz; Miladi, Mahsan; Al-Fdeilat, Abdullah H.; Somogyi, Arpad; Solouki, Touradj

2014-02-01

To date, only a limited number of reports are available on structural variants of multiply-charged b-fragment ions. We report on observed bimodal gas-phase hydrogen/deuterium exchange (HDX) reaction kinetics and patterns for substance P b10 2+ that point to presence of isomeric structures. We also compare HDX reactions, post-ion mobility/collision-induced dissociation (post-IM/CID), and sustained off-resonance irradiation-collision induced dissociation (SORI-CID) of substance P b10 2+ and a cyclic peptide with an identical amino acid (AA) sequence order to substance P b10. The observed HDX patterns and reaction kinetics and SORI-CID pattern for the doubly charged head-to-tail cyclized peptide were different from either of the presumed isomers of substance P b10 2+, suggesting that b10 2+ may not exist exclusively as a head-to-tail cyclized structure. Ultra-high mass measurement accuracy was used to assign identities of the observed SORI-CID fragment ions of substance P b10 2+; over 30 % of the observed SORI-CID fragment ions from substance P b10 2+ had rearranged (scrambled) AA sequences. Moreover, post-IM/CID experiments revealed the presence of two conformer types for substance P b10 2+, whereas only one conformer type was observed for the head-to-tail cyclized peptide. We also show that AA sequence scrambling from CID of doubly-charged b-fragment ions is not unique to substance P b10 2+.

Evidence for sequence scrambling and divergent H/D exchange reactions of doubly-charged isobaric b-type fragment ions.

PubMed

Zekavat, Behrooz; Miladi, Mahsan; Al-Fdeilat, Abdullah H; Somogyi, Arpad; Solouki, Touradj

2014-02-01

To date, only a limited number of reports are available on structural variants of multiply-charged b-fragment ions. We report on observed bimodal gas-phase hydrogen/deuterium exchange (HDX) reaction kinetics and patterns for substance P b10(2+) that point to presence of isomeric structures. We also compare HDX reactions, post-ion mobility/collision-induced dissociation (post-IM/CID), and sustained off-resonance irradiation-collision induced dissociation (SORI-CID) of substance P b10(2+) and a cyclic peptide with an identical amino acid (AA) sequence order to substance P b10. The observed HDX patterns and reaction kinetics and SORI-CID pattern for the doubly charged head-to-tail cyclized peptide were different from either of the presumed isomers of substance P b10(2+), suggesting that b10(2+) may not exist exclusively as a head-to-tail cyclized structure. Ultra-high mass measurement accuracy was used to assign identities of the observed SORI-CID fragment ions of substance P b10(2+); over 30% of the observed SORI-CID fragment ions from substance P b10(2+) had rearranged (scrambled) AA sequences. Moreover, post-IM/CID experiments revealed the presence of two conformer types for substance P b10(2+), whereas only one conformer type was observed for the head-to-tail cyclized peptide. We also show that AA sequence scrambling from CID of doubly-charged b-fragment ions is not unique to substance P b10(2+).

Indole-3-Carbonitriles as DYRK1A Inhibitors by Fragment-Based Drug Design.

PubMed

Meine, Rosanna; Becker, Walter; Falke, Hannes; Preu, Lutz; Loaëc, Nadège; Meijer, Laurent; Kunick, Conrad

2018-01-24

Dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) is a potential drug target because of its role in the development of Down syndrome and Alzheimer's disease. The selective DYRK1A inhibitor 10-iodo-11 H -indolo[3,2- c ]quinoline-6-carboxylic acid (KuFal194), a large, flat and lipophilic molecule, suffers from poor water solubility, limiting its use as chemical probe in cellular assays and animal models. Based on the structure of KuFal194, 7-chloro-1 H -indole-3-carbonitrile was selected as fragment template for the development of smaller and less lipophilic DYRK1A inhibitors. By modification of this fragment, a series of indole-3-carbonitriles was designed and evaluated as potential DYRK1A ligands by molecular docking studies. Synthesis and in vitro assays on DYRK1A and related protein kinases identified novel double-digit nanomolar inhibitors with submicromolar activity in cell culture assays.

Diet and environment 1.2 million years ago revealed through analysis of dental calculus from Europe's oldest hominin at Sima del Elefante, Spain

NASA Astrophysics Data System (ADS)

Hardy, Karen; Radini, Anita; Buckley, Stephen; Blasco, Ruth; Copeland, Les; Burjachs, Francesc; Girbal, Josep; Yll, Riker; Carbonell, Eudald; Bermúdez de Castro, Jose María

2017-02-01

Sima del Elefante, Atapuerca, Spain contains one of the earliest hominin fragments yet known in Europe, dating to 1.2 Ma. Dental calculus from a hominin molar was removed, degraded and analysed to recover entrapped remains. Evidence for plant use at this time is very limited and this study has revealed the earliest direct evidence for foods consumed in the genus Homo. This comprises starchy carbohydrates from two plants, including a species of grass from the Triticeae or Bromideae tribe, meat and plant fibres. All food was eaten raw, and there is no evidence for processing of the starch granules which are intact and undamaged. Additional biographical detail includes fragments of non-edible wood found adjacent to an interproximal groove suggesting oral hygiene activities, while plant fibres may be linked to raw material processing. Environmental evidence comprises spores, insect fragments and conifer pollen grains which are consistent with a forested environment.

Diet and environment 1.2 million years ago revealed through analysis of dental calculus from Europe's oldest hominin at Sima del Elefante, Spain.

PubMed

Hardy, Karen; Radini, Anita; Buckley, Stephen; Blasco, Ruth; Copeland, Les; Burjachs, Francesc; Girbal, Josep; Yll, Riker; Carbonell, Eudald; Bermúdez de Castro, Jose María

2017-02-01

Sima del Elefante, Atapuerca, Spain contains one of the earliest hominin fragments yet known in Europe, dating to 1.2 Ma. Dental calculus from a hominin molar was removed, degraded and analysed to recover entrapped remains. Evidence for plant use at this time is very limited and this study has revealed the earliest direct evidence for foods consumed in the genus Homo. This comprises starchy carbohydrates from two plants, including a species of grass from the Triticeae or Bromideae tribe, meat and plant fibres. All food was eaten raw, and there is no evidence for processing of the starch granules which are intact and undamaged. Additional biographical detail includes fragments of non-edible wood found adjacent to an interproximal groove suggesting oral hygiene activities, while plant fibres may be linked to raw material processing. Environmental evidence comprises spores, insect fragments and conifer pollen grains which are consistent with a forested environment.

Charge Transfer Dissociation of Complex Oligosaccharides: Comparison with Collision-Induced Dissociation and Extreme Ultraviolet Dissociative Photoionization

NASA Astrophysics Data System (ADS)

Ropartz, David; Li, Pengfei; Fanuel, Mathieu; Giuliani, Alexandre; Rogniaux, Hélène; Jackson, Glen P.

2016-10-01

The structural characterization of oligosaccharides still challenges the field of analytical chemistry. Tandem mass spectrometry offers many advantages toward this aim, although the generic fragmentation method (low-energy collision-induced dissociation) shows clear limitations and is often insufficient to retrieve some essential structural information on these molecules. In this work, we present the first application of helium charge transfer dissociation (He-CTD) to characterize the structure of complex oligosaccharides. We compare this method with low-energy collision-induced dissociation and extreme-ultraviolet dissociative photoionization (XUV-DPI), which was shown previously to ensure the successful characterization of complex glycans. Similarly to what could be obtained by XUV-DPI, He-CTD provides a complete description of the investigated structures by producing many informative cross-ring fragments and no ambiguous fragmentation. Unlike XUV-DPI, which is performed at a synchrotron source, He-CTD has the undeniable advantage of being implementable in a conventional benchtop ion trap in a conventional laboratory setting.

Optimization and characterization of a cemented ultimate-storage product

NASA Astrophysics Data System (ADS)

Brunner, H.

1981-12-01

The U- and Pu-containing packaging wastes can be homogeneously cemented after a washing and fragmentation process. Both finely crushed and coarsely fragmented raw wastes yield products with sufficient mechanical stability. The processability limit of the coarsely fragmented raw waste using cement paste or mortar is largely determined by the cellulose content, which is not to exceed 1.3% by weight in the end waste. Of 9 binders studied, the most corrosion-resistant products were obtained with blast-furnace slag cement, whereas poured concrete and Maxit are much less resistant in five-component brine. In the cemented product, hydrolysis of plasticizers (DOP) from plastics (PVC) occurs, leading to release of 2-ethyl-hexanol. This reaction occurs to a much lower degree with blast-furnace slag cement than with all other binders studied. The binder chosen for further tests consists of blast-furnace slag cement, concrete fluidizer and a stabilizer, and is processed at a W/C ratio of 0.43.

In vitro Fab display: a cell-free system for IgG discovery

PubMed Central

Stafford, Ryan L.; Matsumoto, Marissa L.; Yin, Gang; Cai, Qi; Fung, Juan Jose; Stephenson, Heather; Gill, Avinash; You, Monica; Lin, Shwu-Hwa; Wang, Willie D.; Masikat, Mary Rose; Li, Xiaofan; Penta, Kalyani; Steiner, Alex R.; Baliga, Ramesh; Murray, Christopher J.; Thanos, Christopher D.; Hallam, Trevor J.; Sato, Aaron K.

2014-01-01

Selection technologies such as ribosome display enable the rapid discovery of novel antibody fragments entirely in vitro. It has been assumed that the open nature of the cell-free reactions used in these technologies limits selections to single-chain protein fragments. We present a simple approach for the selection of multi-chain proteins, such as antibody Fab fragments, using ribosome display. Specifically, we show that a two-chain trastuzumab (Herceptin) Fab domain can be displayed in a format which tethers either the heavy or light chain to the ribosome while retaining functional antigen binding. Then, we constructed synthetic Fab HC and LC libraries and performed test selections against carcinoembryonic antigen (CEA) and vascular endothelial growth factor (VEGF). The Fab selection output was reformatted into full-length immunoglobulin Gs (IgGs) and directly expressed at high levels in an optimized cell-free system for immediate screening, purification and characterization. Several novel IgGs were identified using this cell-free platform that bind to purified CEA, CEA positive cells and VEGF. PMID:24586053

A fluorescent aptasensor for sensitive analysis oxytetracycline based on silver nanoclusters.

PubMed

Hosseini, Morteza; Mehrabi, Fatemeh; Ganjali, Mohammad Reza; Norouzi, Parviz

2016-11-01

A fluorescent aptasensor for detection of oxytetracycline (OTC) was presented based on fluorescence quenching of DNA aptamer-templated silver nanoclusters (AgNCs). The specific DNA scaffolds with two different nucleotides fragments were used: one was enriched with a cytosine sequence fragment (C12) that could produce DNA-AgNCs via a chemical reduction method, and another was the OTC aptamer fragment that could selectively bind to the OTC antibiotic. Thus, the as-prepared AgNCs could exhibit quenched fluorescence after binding to the target OTC. The fluorescence ratio of the DNA-AgNCs was quenched in a linearly proportional manner to the concentration of the target in the range of 0.5 nM to 100 nM with a detection limit of 0.1 nM. This proposed nanobiosensor was demonstrated to be sensitive, selective, and simple, introducing a viable alternative for rapid determination of toxin OTC in honey and water samples. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

20 years since the introduction of DNA barcoding: from theory to application.

PubMed

Fišer Pečnikar, Živa; Buzan, Elena V

2014-02-01

Traditionally, taxonomic identification has relied upon morphological characters. In the last two decades, molecular tools based on DNA sequences of short standardised gene fragments, termed DNA barcodes, have been developed for species discrimination. The most common DNA barcode used in animals is a fragment of the cytochrome c oxidase (COI) mitochondrial gene, while for plants, two chloroplast gene fragments from the RuBisCo large subunit (rbcL) and maturase K (matK) genes are widely used. Information gathered from DNA barcodes can be used beyond taxonomic studies and will have far-reaching implications across many fields of biology, including ecology (rapid biodiversity assessment and food chain analysis), conservation biology (monitoring of protected species), biosecurity (early identification of invasive pest species), medicine (identification of medically important pathogens and their vectors) and pharmacology (identification of active compounds). However, it is important that the limitations of DNA barcoding are understood and techniques continually adapted and improved as this young science matures.

The thermal history of interplanetary dust particles collected in the Earth's stratosphere

NASA Technical Reports Server (NTRS)

Nier, A. O.; Schlutter, D. J.

1993-01-01

Fragments of 24 individual interplanetary dust particles (IDPs) collected in the Earth's stratosphere were obtained from NASA's Johnson Space Center collection and subjected to pulse-heating sequences to extract He and Ne and to learn about the thermal history of the particles. A motivation for the investigation was to see if the procedure would help distinguish between IDPs of asteroidal and cometary origin. The use of a sequence of short-duration heat pulses to perform the extractions is an improvement over the employment of a step-heating sequence, as was used in a previous investigation. The particles studied were fragments of larger parent IDPs, other fragments of which, in coordinated experiments, are undergoing studies of elemental and mineralogical composition in other laboratories. While the present investigation will provide useful temperature history data for the particles, the relatively large size of the parent IDPs (approximately 40 micrometers in diameter) resulted in high entry deceleration temperatures. This limited the usefulness of the study for distinguishing between particles of asteroidal and cometary origin.

Structural and functional connectivity as a driver of hillslope erosion following disturbance

USDA-ARS?s Scientific Manuscript database

Hydrologic response to rainfall input on fragmented or burnt hillslopes is strongly influenced by the ensuing connectivity of runoff and erosion processes. Yet, cross-scale process connectivity is seldom evaluated in field studies due scale limitations in experimental design. This study quantified...

The Impact of Data Fragmentation on High-Throughput Clinical Phenotyping

ERIC Educational Resources Information Center

Wei, Weiqi

2012-01-01

Subject selection is essential and has become the rate-limiting step for harvesting knowledge to advance healthcare through clinical research. Present manual approaches inhibit researchers from conducting deep and broad studies and drawing confident conclusions. High-throughput clinical phenotyping (HTCP), a recently proposed approach, leverages…

Status of Pituophis ruthveni (Louisiana pine snake)

Treesearch

D. Craig Rudolph; Shirley J. Burgdorf; Richard R. Schaefer; Richard N. Conner; Ricky W. Maxey

2006-01-01

Extensive trapping surveys across the historical range of Pituophis ruthveni (Louisiana Pine Snake) suggest that extant populations are extremely small and limited to remnant patches of suitable habitat in a highly fragmented landscape. Evaluation of habitat at all known historical localities of P. ruthveni documents the widespread...

Secondary Fragmentation of Comet Shoemaker-Levy 9 and the Ramifications for the Progenitor's Breakup in July 1992

NASA Technical Reports Server (NTRS)

Sekanina, Z.; Chodas, P. W.; Yeomans, D. K.

1998-01-01

Comprehensive analysis of discrete events of secondary fragmentation leads to a conceptually new understanding of the process of disintegration of comet Shoemaker-Levy 9. We submit that the jovian tidal forces inflicted extensive cracks throughout the interior of the original nucleus but did not split it apart. The initial disruption was apparently accomplished by stresses exerted on the cracked object by its fast rotation during the early post-perijove period of time. We argue that this disruption was in fact a rapid sequence of episodes during July 1992 that gave birth to the 12 on-train, or primary, fragments: A, C, D, E, G, H, K, L, Q (later Q(sub 1)), R, S, and W. The discrete events of secondary fragmentation, which gave birth to the off-train fragments, are understood in this scenario as stochastic manifestations of the continuing process of progressive disintegration. Of the 13 off-train fragments considered, nine were secondary--B, F, G(sub 2), M, N, P (later P(sub 2) or P(sub 2a)), Q(sub 2), U, and V--and four tertiary (J, P(sub 1), P(sub 2b), and T). The separation parameters of 11 off-train fragments were determined. The vectorial distribution of separation velocities of these fragments shows a strong concentration toward a great circle, unquestionably an effect of the approximately conserved angular momentum of the progenitor comet since the time of its initial disruption. Also apparent is their clumping (except for P(sub 1)) to a segment along the great circle, implying that the fragments were consistently released from one side of their parents, thus explaining for the first time why the off-train fragments preferentially appeared on one side of the nuclear train. In order to obtain a consistent solution, our model requires that the points of separation be on the antisolar side of the parent fragments, where thermal stresses are likely to enhance the effect of rotation. The episodes of secondary fragmentation are found to have nine months after the close encounter with Jupiter in early July 1992, and the separation velocities ranged between 0.36 and 1.7 m/s. The spin-axis position is determined to have been nearly in the jovicentric orbit plane, which rules out the Asphaug-Benz-Solem strengthless aggregate model as a plausible breakup hypothesis. Since the separation velocities are rotational in nature, they cannot substantially exceed the critical limit for centrifugal breakup and offer an estimate for the original nuclear dimensions. The comet's nucleus is found to have been approximately 10 km in diameter and spinning rapidly. With the exception of P(sub 1) and apparently also P(sub 2) and F, no nongravitational deceleration was detected in the motions of the off-train fragments. Serious doubts are cast on continuing appreciable activity of any of these fragments. Indeed, when it was necessary to introduce a deceleration into the equations of motion, the effect appears to have been due to the action of solar radiation pressure on the centroid of centimeter-sized particulates in the disintegrating condensations.

Diffusion-model analysis of pPb and PbPb collisions at LHC energies

NASA Astrophysics Data System (ADS)

Schulz, P.; Wolschin, G.

2018-06-01

We present an analysis of centrality-dependent pseudorapidity distributions of produced charged hadrons in pPb and PbPb collisions at the Large Hadron Collider (LHC) energy of s NN = 5.02 TeV, and of minimum-bias pPb collisions at 8.16 TeV within the non-equilibrium-statistical relativistic diffusion model (RDM). In a three-source approach, the role of the fragmentation sources is emphasized. Together with the Jacobian transformation from rapidity to pseudorapidity and the limiting fragmentation conjecture, these are essential for modeling the centrality dependence. For central PbPb collisions, a prediction at the projected FCC energy of s NN = 39 TeV is made.

Modulation-frequency encoded multi-color fluorescent DNA analysis in an optofluidic chip.

PubMed

Dongre, Chaitanya; van Weerd, Jasper; Besselink, Geert A J; Vazquez, Rebeca Martinez; Osellame, Roberto; Cerullo, Giulio; van Weeghel, Rob; van den Vlekkert, Hans H; Hoekstra, Hugo J W M; Pollnau, Markus

2011-02-21

We introduce a principle of parallel optical processing to an optofluidic lab-on-a-chip. During electrophoretic separation, the ultra-low limit of detection achieved with our set-up allows us to record fluorescence from covalently end-labeled DNA molecules. Different sets of exclusively color-labeled DNA fragments-otherwise rendered indistinguishable by spatio-temporal coincidence-are traced back to their origin by modulation-frequency-encoded multi-wavelength laser excitation, fluorescence detection with a single ultrasensitive, albeit color-blind photomultiplier, and Fourier analysis decoding. As a proof of principle, fragments obtained by multiplex ligation-dependent probe amplification from independent human genomic segments, associated with genetic predispositions to breast cancer and anemia, are simultaneously analyzed.

Limiting Size of Monolayer Graphene Flakes Grown on Silicon Carbide or via Chemical Vapor Deposition on Different Substrates

NASA Astrophysics Data System (ADS)

Alekseev, N. I.

2018-05-01

The maximum size of homogeneous monolayer graphene flakes that form during the high-temperature evaporation of silicon from a surface of SiC or during graphene synthesis via chemical vapor deposition is estimated, based on the theoretical calculations developed in this work. Conditions conducive to the fragmentation of a monolayer graphene sheet to form discrete fragments or terrace-type structures in which excess energy due to dangling bonds at the edges is compensated for by the lack of internal stress are indentified and described. The results from calculations for the sizes of graphene structures are compared with experimental findings for the most successful graphene syntheses reported in the literature.

Restriction fragment length polymorphisms for growth hormone, prolactin, osteonectin, alpha crystallin, gamma crystallin, fibronectin and 21-steroid hydroxylase in cattle.

PubMed

Theilmann, J L; Skow, L C; Baker, J F; Womack, J E

1989-01-01

Genomic DNAs from animals representing six breeds of cattle (Angus, Brahman, Hereford, Holstein, Jersey and Texas Longhorn) were screened with cloned gene probes in a search for restriction fragment length polymorphisms (RFLPs). Eleven RFLPs were identified using seven different probes: growth hormone, prolactin, osteonectin, alpha A-crystallin, gamma crystallin, fibronectin and 21-steroid hydroxylase. The frequencies of the alleles identified by each probe were calculated and compared in a limited sampling of the six bovine breeds. These polymorphisms greatly enhance the pool of immunogenetic, biochemical and molecular markers available in cattle for linkage analysis, testing of parentage, and distinction of breeds.

Coagulation-Fragmentation Model for Animal Group-Size Statistics

NASA Astrophysics Data System (ADS)

Degond, Pierre; Liu, Jian-Guo; Pego, Robert L.

2017-04-01

We study coagulation-fragmentation equations inspired by a simple model proposed in fisheries science to explain data for the size distribution of schools of pelagic fish. Although the equations lack detailed balance and admit no H-theorem, we are able to develop a rather complete description of equilibrium profiles and large-time behavior, based on recent developments in complex function theory for Bernstein and Pick functions. In the large-population continuum limit, a scaling-invariant regime is reached in which all equilibria are determined by a single scaling profile. This universal profile exhibits power-law behavior crossing over from exponent -2/3 for small size to -3/2 for large size, with an exponential cutoff.

Structure and physical characteristics of pumice from the climactic eruption of Mount Mazama (Crater Lake), Oregon

USGS Publications Warehouse

Klug, C.; Cashman, K.; Bacon, C.

2002-01-01

The vesicularity, permeability, and structure of pumice clasts provide insight into conditions of vesiculation and fragmentation during Plinian fall and pyroclastic flow-producing phases of the ???7,700 cal. year B.P. climactic eruption of Mount Mazama (Crater Lake), Oregon. We show that bulk properties (vesicularity and permeability) can be correlated with internal textures and that the clast structure can be related to inferred changes in eruption conditions. The vesicularity of all pumice clasts is 75-88%, with >90% interconnected pore volume. However, pumice clasts from the Plinian fall deposits exhibit a wider vesicularity range and higher volume percentage of interconnected vesicles than do clasts from pyroclastic-flow deposits. Pumice permeabilities also differ between the two clast types, with pumice from the fall deposit having higher minimum permeabilities (???5??10-13 m2) and a narrower permeability range (5-50??10-13 m2) than clasts from pyroclastic-flow deposits (0.2-330??10-13 m2). The observed permeability can be modeled to estimate average vesicle aperture radii of 1-5 ??m for the fall deposit clasts and 0.25-1 ??m for clasts from the pyroclastic flows. High vesicle number densities (???109 cm-3) in all clasts suggest that bubble nucleation occured rapidly and at high supersaturations. Post-nucleation modifications to bubble populations include both bubble growth and coalescence. A single stage of bubble nucleation and growth can account for 35-60% of the vesicle population in clasts from the fall deposits, and 65-80% in pumice from pyroclastic flows. Large vesicles form a separate population which defines a power law distribution with fractal dimension D=3.3 (range 3.0-3.5). The large D.value, coupled with textural evidence, suggests that the large vesicles formed primarily by coalescence. When viewed together, the bulk properties (vesicularity, permeability) and textural characteristics of all clasts indicate rapid bubble nucleation followed by bubble growth, coalescence and permeability development. This sequence of events is best explained by nucleation in response to a downward-propagating decompression wave, followed by rapid bubble growth and coalescence prior to magma disruption by fragmentation. The heterogeneity of vesicle sizes and shapes, and the absence of differential expansion across individual clasts, suggest that post-fragmentation expansion played a limited role in the development of pumice structure. The higher vesicle number densities and lower permeabilities of pyroclastic-flow clasts indicate limited coalescence and suggest that fragmentation occurred shortly after decompression. Either increased eruption velocities or increased depth of fragmentation accompanying caldera collapse could explain compression of the pre-fragmentation vesiculation interval.

A multi-particle crushing apparatus for studying rock fragmentation due to repeated impacts

NASA Astrophysics Data System (ADS)

Huang, S.; Mohanty, B.; Xia, K.

2017-12-01

Rock crushing is a common process in mining and related operations. Although a number of particle crushing tests have been proposed in the literature, most of them are concerned with single-particle crushing, i.e., a single rock sample is crushed in each test. Considering the realistic scenario in crushers where many fragments are involved, a laboratory crushing apparatus is developed in this study. This device consists of a Hopkinson pressure bar system and a piston-holder system. The Hopkinson pressure bar system is used to apply calibrated dynamic loads to the piston-holder system, and the piston-holder system is used to hold rock samples and to recover fragments for subsequent particle size analysis. The rock samples are subjected to three to seven impacts under three impact velocities (2.2, 3.8, and 5.0 m/s), with the feed size of the rock particle samples limited between 9.5 and 12.7 mm. Several key parameters are determined from this test, including particle size distribution parameters, impact velocity, loading pressure, and total work. The results show that the total work correlates well with resulting fragmentation size distribution, and the apparatus provides a useful tool for studying the mechanism of crushing, which further provides guidelines for the design of commercial crushers.

Genetic analysis reveals demographic fragmentation of grizzly bears yielding vulnerably small populations

PubMed Central

Proctor, Michael F; McLellan, Bruce N; Strobeck, Curtis; Barclay, Robert M.R

2005-01-01

Ecosystem conservation requires the presence of native carnivores, yet in North America, the distributions of many larger carnivores have contracted. Large carnivores live at low densities and require large areas to thrive at the population level. Therefore, if human-dominated landscapes fragment remaining carnivore populations, small and demographically vulnerable populations may result. Grizzly bear range contraction in the conterminous USA has left four fragmented populations, three of which remain along the Canada–USA border. A tenet of grizzly bear conservation is that the viability of these populations requires demographic linkage (i.e. inter-population movement of both sexes) to Canadian bears. Using individual-based genetic analysis, our results suggest this demographic connection has been severed across their entire range in southern Canada by a highway and associated settlements, limiting female and reducing male movement. Two resulting populations are vulnerably small (≤100 animals) and one of these is completely isolated. Our results suggest that these trans-border bear populations may be more threatened than previously thought and that conservation efforts must expand to include international connectivity management. They also demonstrate the ability of genetic analysis to detect gender-specific demographic population fragmentation in recently disturbed systems, a traditionally intractable yet increasingly important ecological measurement worldwide. PMID:16243699

Label-enhanced surface plasmon resonance applied to label-free interaction analysis of small molecules and fragments.

PubMed

Eng, Lars; Nygren-Babol, Linnéa; Hanning, Anders

2016-10-01

Surface plasmon resonance (SPR) is a well-established method for studying interactions between small molecules and biomolecules. In particular, SPR is being increasingly applied within fragment-based drug discovery; however, within this application area, the limited sensitivity of SPR may constitute a problem. This problem can be circumvented by the use of label-enhanced SPR that shows a 100-fold higher sensitivity as compared with conventional SPR. Truly label-free interaction data for small molecules can be obtained by applying label-enhanced SPR in a surface competition assay format. The enhanced sensitivity is accompanied by an increased specificity and inertness toward disturbances (e.g., bulk refractive index disturbances). Label-enhanced SPR can be used for fragment screening in a competitive assay format; the competitive format has the added advantage of confirming the specificity of the molecular interaction. In addition, label-enhanced SPR extends the accessible kinetic regime of SPR to the analysis of very fast fragment binding kinetics. In this article, we demonstrate the working principles and benchmark the performance of label-enhanced SPR in a model system-the interaction between carbonic anhydrase II and a number of small-molecule sulfonamide-based inhibitors. Copyright © 2016 Elsevier Inc. All rights reserved.

Target immobilization as a strategy for NMR-based fragment screening: comparison of TINS, STD, and SPR for fragment hit identification.

PubMed

Kobayashi, Masakazu; Retra, Kim; Figaroa, Francis; Hollander, Johan G; Ab, Eiso; Heetebrij, Robert J; Irth, Hubertus; Siegal, Gregg

2010-09-01

Fragment-based drug discovery (FBDD) has become a widely accepted tool that is complementary to high-throughput screening (HTS) in developing small-molecule inhibitors of pharmaceutical targets. Because a fragment campaign can only be as successful as the hit matter found, it is critical that the first stage of the process be optimized. Here the authors compare the 3 most commonly used methods for hit discovery in FBDD: high concentration screening (HCS), solution ligand-observed nuclear magnetic resonance (NMR), and surface plasmon resonance (SPR). They selected the commonly used saturation transfer difference (STD) NMR spectroscopy and the proprietary target immobilized NMR screening (TINS) as representative of the array of possible NMR methods. Using a target typical of FBDD campaigns, the authors find that HCS and TINS are the most sensitive to weak interactions. They also find a good correlation between TINS and STD for tighter binding ligands, but the ability of STD to detect ligands with affinity weaker than 1 mM K(D) is limited. Similarly, they find that SPR detection is most suited to ligands that bind with K(D) better than 1 mM. However, the good correlation between SPR and potency in a bioassay makes this a good method for hit validation and characterization studies.

Genetic analysis reveals demographic fragmentation of grizzly bears yielding vulnerably small populations.

PubMed

Proctor, Michael F; McLellan, Bruce N; Strobeck, Curtis; Barclay, Robert M R

2005-11-22

Ecosystem conservation requires the presence of native carnivores, yet in North America, the distributions of many larger carnivores have contracted. Large carnivores live at low densities and require large areas to thrive at the population level. Therefore, if human-dominated landscapes fragment remaining carnivore populations, small and demographically vulnerable populations may result. Grizzly bear range contraction in the conterminous USA has left four fragmented populations, three of which remain along the Canada-USA border. A tenet of grizzly bear conservation is that the viability of these populations requires demographic linkage (i.e. inter-population movement of both sexes) to Canadian bears. Using individual-based genetic analysis, our results suggest this demographic connection has been severed across their entire range in southern Canada by a highway and associated settlements, limiting female and reducing male movement. Two resulting populations are vulnerably small (< or =100 animals) and one of these is completely isolated. Our results suggest that these trans-border bear populations may be more threatened than previously thought and that conservation efforts must expand to include international connectivity management. They also demonstrate the ability of genetic analysis to detect gender-specific demographic population fragmentation in recently disturbed systems, a traditionally intractable yet increasingly important ecological measurement worldwide.

On fragmentation of turbulent self-gravitating discs in the long cooling time regime

NASA Astrophysics Data System (ADS)

Rice, Ken; Nayakshin, Sergei

2018-03-01

It has recently been suggested that in the presence of driven turbulence discs may be much less stable against gravitational collapse than their non-turbulent analogues, due to stochastic density fluctuations in turbulent flows. This mode of fragmentation would be especially important for gas giant planet formation. Here, we argue, however, that stochastic density fluctuations due to turbulence do not enhance gravitational instability and disc fragmentation in the long cooling time limit appropriate for planet forming discs. These fluctuations evolve adiabatically and dissipate away by decompression faster than they could collapse. We investigate these issues numerically in two dimensions via shearing box simulations with driven turbulence and also in three dimensions with a model of instantaneously applied turbulent velocity kicks. In the former setting turbulent driving leads to additional disc heating that tends to make discs more, rather than less, stable to gravitational instability. In the latter setting, the formation of high-density regions due to convergent velocity kicks is found to be quickly followed by decompression, as expected. We therefore conclude that driven turbulence does not promote disc fragmentation in protoplanetary discs and instead tends to make the discs more stable. We also argue that sustaining supersonic turbulence is very difficult in discs that cool slowly.

Monovalent IgG4 molecules

PubMed Central

Wilkinson, Ian C.; Fowler, Susan B.; Machiesky, LeeAnn; Miller, Kenneth; Hayes, David B.; Adib, Morshed; Her, Cheng; Borrok, M. Jack; Tsui, Ping; Burrell, Matthew; Corkill, Dominic J.; Witt, Susanne; Lowe, David C.; Webster, Carl I.

2013-01-01

Antibodies have become the fastest growing class of biological therapeutics, in part due to their exquisite specificity and ability to modulate protein-protein interactions with a high biological potency. The relatively large size and bivalency of antibodies, however, limits their use as therapeutics in certain circumstances. Antibody fragments, such as single-chain variable fragments and antigen binding-fragments, have emerged as viable alternatives, but without further modifications these monovalent formats have reduced terminal serum half-lives because of their small size and lack of an Fc domain, which is required for FcRn-mediated recycling. Using rational engineering of the IgG4 Fc domain to disrupt key interactions at the CH3-CH3 interface, we identified a number of point mutations that abolish Fc dimerization and created half-antibodies, a novel monovalent antibody format that retains a monomeric Fc domain. Introduction of these mutations into an IgG1 framework also led to the creation of half-antibodies. These half-antibodies were shown to be soluble, thermodynamically stable and monomeric, characteristics that are favorable for use as therapeutic proteins. Despite significantly reduced FcRn binding in vitro, which suggests that avidity gains in a dimeric Fc are critical to optimal FcRn binding, this format demonstrated an increased terminal serum half-life compared with that expected for most alternative antibody fragments. PMID:23567207

Early Childhood Education in Botswana: A Case of Fragmented "Fits"

ERIC Educational Resources Information Center

Maunganidze, Langtone; Tsamaase, Marea

2014-01-01

Scholarly and institutional research on early childhood education is not a new phenomenon, but what is perhaps limited is extant literature that particularly focuses on revisiting current practices and their "dialogue" with the ever-changing environment that prevails especially in an African setting. The practices of providers of early…

14 CFR 431.43 - Reusable launch vehicle mission operational requirements and restrictions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... debris generation will not result from conversion of energy sources into energy that fragments the vehicle or its payload. Energy sources include, but are not limited to, chemical, pneumatic, and kinetic energy; and (4) Vehicle safety operations personnel shall adhere to the following work and rest standards...

14 CFR 431.43 - Reusable launch vehicle mission operational requirements and restrictions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... debris generation will not result from conversion of energy sources into energy that fragments the vehicle or its payload. Energy sources include, but are not limited to, chemical, pneumatic, and kinetic energy; and (4) Vehicle safety operations personnel shall adhere to the following work and rest standards...

14 CFR 431.43 - Reusable launch vehicle mission operational requirements and restrictions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... debris generation will not result from conversion of energy sources into energy that fragments the vehicle or its payload. Energy sources include, but are not limited to, chemical, pneumatic, and kinetic energy; and (4) Vehicle safety operations personnel shall adhere to the following work and rest standards...

14 CFR 431.43 - Reusable launch vehicle mission operational requirements and restrictions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... debris generation will not result from conversion of energy sources into energy that fragments the vehicle or its payload. Energy sources include, but are not limited to, chemical, pneumatic, and kinetic energy; and (4) Vehicle safety operations personnel shall adhere to the following work and rest standards...

14 CFR 431.43 - Reusable launch vehicle mission operational requirements and restrictions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... debris generation will not result from conversion of energy sources into energy that fragments the vehicle or its payload. Energy sources include, but are not limited to, chemical, pneumatic, and kinetic energy; and (4) Vehicle safety operations personnel shall adhere to the following work and rest standards...

Distinctive and Complementary MS2 Fragmentation Characteristics for Identification of Sulfated Sialylated N-Glycopeptides by nanoLC-MS/MS Workflow

NASA Astrophysics Data System (ADS)

Kuo, Chu-Wei; Guu, Shih-Yun; Khoo, Kay-Hooi

2018-04-01

High sensitivity identification of sulfated glycans carried on specific sites of glycoproteins is an important requisite for investigation of molecular recognition events involved in diverse biological processes. However, aiming for resolving site-specific glycosylation of sulfated glycopeptides by direct LC-MS2 sequencing is technically most challenging. Other than the usual limiting factors such as lower abundance and ionization efficiency compared to analysis of non-glycosylated peptides, confident identification of sulfated glycopeptides among the more abundant non-sulfated glycopeptides requires additional considerations in the selective enrichment and detection strategies. Metal oxide has been applied to enrich phosphopeptides and sialylated glycopeptides, but its use to capture sulfated glycopeptides has not been investigated. Likewise, various complementary MS2 fragmentation modes have yet to be tested against sialylated and non-sialylated sulfoglycopeptides due to limited appropriate sample availability. In this study, we have investigated the feasibility of sequencing tryptic sulfated N-glycopeptide and its MS2 fragmentation characteristics by first optimizing the enrichment methods to allow efficient LC-MS detection and MS2 analysis by a combination of CID, HCD, ETD, and EThcD on hybrid and tribrid Orbitrap instruments. Characteristic sulfated glyco-oxonium ions and direct loss of sulfite from precursors were detected as evidences of sulfate modification. It is anticipated that the technical advances demonstrated in this study would allow a feasible extension of our sulfoglycomic analysis to sulfoglycoproteomics. [Figure not available: see fulltext.

Minimal debridement or simple wound closure as the only surgical treatment in war victims with low-velocity penetrating head injuries. Indications and management protocol based upon more than 8 years follow-up of 99 cases from Iran-Iraq conflict.

PubMed

Amirjamshidi, Abbass; Abbassioun, Kazem; Rahmat, Hamid

2003-08-01

During military conflicts, surviving victims traditionally underwent large enough craniectomy or craniotomy to achieve extensive debridement of the in-driven bone, shell fragments, and debris. Meanwhile, the completely as well as the partially devitalized adjacent brain tissue would also be debrided. This might have led to additional neurologic deficit or lesser degree of recovery of functions. We studied the efficacy and safety of much more limited or even absence of surgical intervention in a selected group of wartime head-wounded patients. Among 1,150 war victims with penetrating head wounds referred to two major centers affiliated with Tehran University of Medical Sciences during the 8-year period spanning the Iran-Iraq conflict, there were 191 head-wounded patients who underwent either no surgical intervention or a very limited debridement. We have attempted to follow up as closely as possible for more than 8 years. Ninety-nine of them fulfilled the criteria to be included in this series. Close follow-up for development of cerebral infections, functional recovery, or development of new neurologic deficits and epilepsy were performed. At least 3 contrast-enhanced CT scans were taken for each patient. All patients were victims of the battle-field areas with low- to moderate-velocity missile or shell fragment injury. The Glasgow Coma Scale (GCS) scores of the patients were between 8-14 when visited by the first attending neurosurgery staff. The factors considered to be effective in the outcome analysis were the entry point, the number of the in-driven bone fragments, the retained shell fragments, the brain compartments affected, and the paranasal sinus or skull base involvement with or without cerebrospinal fluid (CSF) fistula. There were 13 unilobar, 44 bilobar, 4 trilobar, 38 transventricular, and 3 skull base lesions. The number of retained metal fragments varied between 1 to 6. The number of retained bone fragments varied between 1 to 5 in 73 victims and more than five chips in other cases. The outcome was good (back to work) in 90 patients and poor (severely disabled) in 4. Five patients died because of severity of brain damage and meningitis. The effect of the different enumerated variables upon the outcome was measured using chi(2) and Fisher exact test, which was nonsignificant for all of the variables except for the orbitofacial entry point of injury (p = 0.00006). This study seems to indicate that not only is reoperation for retained bone or shell fragments unnecessary, but surgeons having modern neuroradiological instruments available to detect the proper explanation for changes in GCS of the patients may decide not to proceed with any surgical intervention in a good number of patients. As a result of the "no surgical treatment protocol," one can preserve cerebral tissue without exposing the patient to undue risk of seizures, infection, or death by leaving behind some or all the in-driven bone and shell fragments.

Quantifying edge effect extent and its impacts on carbon stocks across a degraded landscape in the Amazon using airborne lidar.

NASA Astrophysics Data System (ADS)

dos-Santos, M. N.; Keller, M.; Morton, D. C.; Longo, M.; Scaranello, M. A., Sr.; Pinagé, E. R.; Correa Pabon, R.

2017-12-01

Ongoing tropical forest degradation and forest fragmentation increases forest edge area. Forest edges experience hotter, drier, and windier conditions and greater exposure to fires compared to interior areas, which elevate rates of tree mortality. Previous studies have suggested that forests within 100 m from the edge may lose 36% of biomass during the first two decades following fragmentation, although such estimates are based on a limited number of experimental plots. Degraded forests behave differently from intact forests and quantifying edge effect extension in a degraded forest landscape is more challenging compared to experimental studies. To overcome these limitations, we used airborne lidar data to quantify changes in forest structure near 91 edges in a heavily degraded tropical forest in Paragominas Municipality, eastern Brazilian Amazon. Paragominas was a center of timber production in the 1990s. Today, the landscape is a mosaic of different agricultural uses, degraded, secondary and unmanaged forests. A total of 3000 ha of high density (mean density of 17.9 points/m2) lidar data were acquired in August/September 2013 and June/July 2014 over 30 transects (200 x 5000m), systematically distributed over the study area, using the Optech Orion M-200 laser scanning system. We adopted lidar-measured forest heights as the edge effect criteria and found that mean extent of edge effect was highly variable across degraded forests (150 ± 354m) and secondary forest fragments (265 ± 365m). We related the extent of forest edges to the historical disturbances identified in Landsat imagery since 1984. Contrary to previous studies, we found that carbon stocks along forest edges were not significantly lower than forest core biomass when edges were defined by previously estimated range of 100 and 300m. In frontier forests, ecological edge effect may be masked by the cumulative impact of historic forest degradation - an anthropogenic edge effect that extends beyond the scale of changes in forest microclimate from fragmentation.

Landscape attributes as drivers of the geographical variation in density of Sapajus nigritus Kerr, 1792, a primate endemic to the Atlantic Forest

NASA Astrophysics Data System (ADS)

Hendges, Carla D.; Melo, Geruza L.; Gonçalves, Alberto S.; Cerezer, Felipe O.; Cáceres, Nilton C.

2017-10-01

Neotropical primates are among the most well studied forest mammals concerning their population densities. However, few studies have evaluated the factors that influence the spatial variation in the population density of primates, which limits the possibility of inferences towards this animal group, especially at the landscape-level. Here, we compiled density data of Sapajus nigritus from 21 forest patches of the Brazilian Atlantic Forest. We tested the effects of climatic variables (temperature, precipitation), landscape attributes (number of patches, mean inter-patch isolation distance, matrix modification index) and patch size on the population density using linear models and the Akaike information criterion. Our findings showed that the density of S. nigritus is influenced by landscape attributes, particularly by fragmentation and matrix modification. Overall, moderately fragmented landscapes and those surrounded by matrices with intermediate indexes of temporal modification (i.e., crop plantations, forestry) are related to high densities of this species. These results support the assumptions that ecologically flexible species respond positively to forest fragmentation. However, the non-linear relationship between S. nigritus density and number of patches suggests that even the species that are most tolerant to forest cover changes seem to respond positively only at an intermediate level of habitat fragmentation, being dependent of both a moderate degree of forest cover and a high quality matrix. The results we found here can be a common response to fragmentation for those forest dweller species that are able to use the matrix as complementary foraging sites.

Gene flow and pathogen transmission among bobcats (Lynx rufus) in a fragmented urban landscape

USGS Publications Warehouse

Lee, Justin S.; Ruell, Emily W.; Boydston, Erin E.; Lyren, Lisa M.; Alonso, Robert S.; Troyer, Jennifer L.; Crooks, Kevin R.; VandeWoude, Sue

2012-01-01

Urbanization can result in the fragmentation of once contiguous natural landscapes into a patchy habitat interspersed within a growing urban matrix. Animals living in fragmented landscapes often have reduced movement among habitat patches because of avoidance of intervening human development, which potentially leads to both reduced gene flow and pathogen transmission between patches. Mammalian carnivores with large home ranges, such as bobcats (Lynx rufus), may be particularly sensitive to habitat fragmentation. We performed genetic analyses on bobcats and their directly transmitted viral pathogen, feline immunodeficiency virus (FIV), to investigate the effects of urbanization on bobcat movement. We predicted that urban development, including major freeways, would limit bobcat movement and result in genetically structured host and pathogen populations. We analysed molecular markers from 106 bobcats and 19 FIV isolates from seropositive animals in urban southern California. Our findings indicate that reduced gene flow between two primary habitat patches has resulted in genetically distinct bobcat subpopulations separated by urban development including a major highway. However, the distribution of genetic diversity among FIV isolates determined through phylogenetic analyses indicates that pathogen genotypes are less spatially structured--exhibiting a more even distribution between habitat fragments. We conclude that the types of movement and contact sufficient for disease transmission occur with enough frequency to preclude structuring among the viral population, but that the bobcat population is structured owing to low levels of effective bobcat migration resulting in gene flow. We illustrate the utility in using multiple molecular markers that differentially detect movement and gene flow between subpopulations when assessing connectivity.

An Efficient Method to Evaluate Intermolecular Interaction Energies in Large Systems Using Overlapping Multicenter ONIOM and the Fragment Molecular Orbital Method

PubMed Central

Asada, Naoya; Fedorov, Dmitri G.; Kitaura, Kazuo; Nakanishi, Isao; Merz, Kenneth M.

2012-01-01

We propose an approach based on the overlapping multicenter ONIOM to evaluate intermolecular interaction energies in large systems and demonstrate its accuracy on several representative systems in the complete basis set limit at the MP2 and CCSD(T) level of theory. In the application to the intermolecular interaction energy between insulin dimer and 4′-hydroxyacetanilide at the MP2/CBS level, we use the fragment molecular orbital method for the calculation of the entire complex assigned to the lowest layer in three-layer ONIOM. The developed method is shown to be efficient and accurate in the evaluation of the protein-ligand interaction energies. PMID:23050059

PCR-based detection of a rare linear DNA in cell culture.

PubMed

Saveliev, Sergei V.

2002-11-11

The described method allows for detection of rare linear DNA fragments generated during genomic deletions. The predicted limit of the detection is one DNA molecule per 10(7) or more cells. The method is based on anchor PCR and involves gel separation of the linear DNA fragment and chromosomal DNA before amplification. The detailed chemical structure of the ends of the linear DNA can be defined with the use of additional PCR-based protocols. The method was applied to study the short-lived linear DNA generated during programmed genomic deletions in a ciliate. It can be useful in studies of spontaneous DNA deletions in cell culture or for tracking intracellular modifications at the ends of transfected DNA during gene therapy trials.

PCR-based detection of a rare linear DNA in cell culture

PubMed Central

2002-01-01

The described method allows for detection of rare linear DNA fragments generated during genomic deletions. The predicted limit of the detection is one DNA molecule per 107 or more cells. The method is based on anchor PCR and involves gel separation of the linear DNA fragment and chromosomal DNA before amplification. The detailed chemical structure of the ends of the linear DNA can be defined with the use of additional PCR-based protocols. The method was applied to study the short-lived linear DNA generated during programmed genomic deletions in a ciliate. It can be useful in studies of spontaneous DNA deletions in cell culture or for tracking intracellular modifications at the ends of transfected DNA during gene therapy trials. PMID:12734566

DOE Office of Scientific and Technical Information (OSTI.GOV)

Horn, Paul R., E-mail: prhorn@berkeley.edu; Mao, Yuezhi; Head-Gordon, Martin, E-mail: mhg@cchem.berkeley.edu

In energy decomposition analysis of Kohn-Sham density functional theory calculations, the so-called frozen (or pre-polarization) interaction energy contains contributions from permanent electrostatics, dispersion, and Pauli repulsion. The standard classical approach to separate them suffers from several well-known limitations. We introduce an alternative scheme that employs valid antisymmetric electronic wavefunctions throughout and is based on the identification of individual fragment contributions to the initial supersystem wavefunction as determined by an energetic optimality criterion. The density deformations identified with individual fragments upon formation of the initial supersystem wavefunction are analyzed along with the distance dependence of the new and classical terms formore » test cases that include the neon dimer, ammonia borane, water-Na{sup +}, water-Cl{sup −}, and the naphthalene dimer.« less

The value of endomyocardial biopsy in diagnosis and guiding therapy.

PubMed

Khan, Tayyaba; Selvakumar, Dinesh; Trivedi, Siddharth; Rao, Karan; Harapoz, Mehmet; Thiagalingam, Aravinda; Denniss, A Robert; Varikatt, Winny

2017-12-01

Endomyocardial biopsy (EMB) is a highly-specialised procedure that is associated with some controversy as to its diagnostic role due to its inconsistency in diagnosing a wide variety of cardiac diseases. Given the advances and sophistication in echocardiography and cardiac magnetic resonance imaging (MRI), the vast majority of cardiac diseases can be diagnosed by these non-invasive procedures. Under-sampling and the fact that biopsy site is limited to the right side of the interventricular septum further limits its value. In spite of all these limitations, there still remains a group of pathological conditions that require biopsy for a conclusive diagnosis such as myocarditis, amyloidosis, sarcoidosis and giant cell myocarditis. Correct patient selection and the quantity of tissue samples impart a significant influence on the accuracy of the diagnosis, and thus the value of EMB is variable for each patient. The purpose of this study was to evaluate the role of EMB in patient care, through its ability to either change clinical diagnosis or alter patient management. Our study was based in a single teaching centre. An audit of cardiac biopsies performed over a 10 year period identified 250 patients. We assessed indications, histology, electron microscopic findings, final clinical diagnosis and how they influenced patient management. A definite diagnosis on histology was given in 44 of 250 patients (17.6%). Non-specific findings were observed in the remaining 206 patients (82.4%). Histology influenced patient management in 73 (29.2%) patients. Histological examination in the remaining 177 biopsies (70.8%) did not provide a definite diagnosis or influence patient management. It was additionally found that the number of tissue fragments sampled has significant impact on diagnostic accuracy. A more accurate diagnosis of 45% was obtained when ≥5 fragments were sampled, as compared to 1-3 fragments where accuracy dropped to 20%. Our study indicated that sampling for electron microscopy has very limited value. We found that of 245 biopsies sampled for electron microscopy, only three biopsies (1.2%) had diagnostically useful findings. In our institution procedure related complications were observed in 7 of 250 patients (2.8%). The diagnostic value of EMB is important but limited. Strict triaging of patients according to clinical suspicion and adequate sampling of tissue may increase useful diagnostic information. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

Integrated Services for Frail Elders (SIPA): A Trial of a Model for Canada

ERIC Educational Resources Information Center

Beland, Francois; Bergman, Howard; Lebel, Paule; Dallaire, Luc; Fletcher, John; Contandriopoulos, Andre-Pierre; Solidage, Tousignant Pierre

2006-01-01

The complex formed by chronic illness, episodes of acute illness, physiological disabilities, functional limitations, and cognitive problems is prevalent among frail elderly persons. These individuals rely on assistance from social and health care programs, which in Canada are still fragmented. SIPA is an integrated service model based on…

Fostering Knowledge and Skills to Teach for Diversity: "There Is Nothing so Practical as a Good Theory"

ERIC Educational Resources Information Center

Marchel, Carol A.; Green, Susan K

2014-01-01

Increased use of field-based teacher preparation offers important opportunities to develop skills with diverse learners. However, limited focus on theoretical content restricts understanding and generalization of well-proven theoretical approaches, resulting in fragmented field applications unlikely to result in broad application. Inspired by Kurt…

Large landscape conservation-synthetic and real-world datasets

Treesearch

Bistra Dilkina; Katherine Lai; Ronan Le Bras; Yexiang Xue; Carla P. Gomes; Ashish Sabharwal; Jordan Suter; Kevin S. McKelvey; Michael K. Schwartz; Claire Montgomery

2013-01-01

Biodiversity underpins ecosystem goods and services and hence protecting it is key to achieving sustainability. However, the persistence of many species is threatened by habitat loss and fragmentation due to human land use and climate change. Conservation efforts are implemented under very limited economic resources, and therefore designing scalable, cost-efficient and...

Development of single chain variable fragment (scFv) antibodies against Xylella fastidiosa subsp. pauca by phage display

USDA-ARS?s Scientific Manuscript database

Xylella fastidiosa is a member of the gamma proteobacteria. It is fastidious, insect-vectored and xylem-limited and causes a variety of diseases, some severe, on a wide range of economically important perennial crops, including grape and citrus. Xylella fastidiosa subsp pauca causes citrus variegat...

Structural and functional connectivity as a driver of hillslope erosion following disturbance

Treesearch

C. Jason Williams; Frederick B. Pierson; Pete Robichaud; Osama Z. Al-Hamdan; Jan Boll; Eva K. Strand

2016-01-01

Hydrologic response to rainfall on fragmented or burnt hillslopes is strongly influenced by the ensuing connectivity of runoff and erosion processes. Yet cross-scale process connectivity is seldom evaluated in field studies owing to scale limitations in experimental design. This study quantified surface susceptibility and hydrologic response across point to...

Distinguishing stressors acting on landbird communities in an urbanizing environment

Treesearch

Matthew D. Schlesinger; Patricia N. Manley; Marcel Holyoak

2008-01-01

Urbanization has profound influences on ecological communities, but our understanding of causal mechanisms is limited by a lack of attention to its component stressors. Published research suggests that at landscape scales, habitat loss and fragmentation are the major drivers of community change, whereas at local scales, human activity and vegetation management are the...

Highways and habitat: managing habitat connectivity and landscape permeability for wildlife.

Treesearch

Jonathan Thompson

2006-01-01

Millions of miles of highway crisscross the United States . Highways fragment the landscape, affecting the distribution of animal populations and limiting the ability of individuals to disperse between those populations. Moreover, animal-vehicle collisions are a serious hazard to wildlife, not to mention people.Researchers at the PNW Research Station in...

Mapping and QTL analysis of drought tolerance in a spring wheat population using AFLP and DArt markers

USDA-ARS?s Scientific Manuscript database

Water availability is commonly the most limiting factor to crop production. This study was conducted to map quantitative trait loci (QTL) involved in drought tolerance in wheat (Triticum aestivum L.) to enable their use for marker assisted selection (MAS) in breeding. Using amplified fragment leng...

Distant views and local realities: The limits of global assessments to restore the fragmented phosphorus cycle

USDA-ARS?s Scientific Manuscript database

With more sophisticated data compilation and analytical capabilities, the evolution of “big data” analysis has occurred rapidly. We examine the meta-analysis of “big data” representing phosphorus (P) flows and stocks in global agriculture and address the need to consider local nuances of farm operat...

Seed dispersal and seedling establishment of Sarracenia purpurea (Sarraceniaceae).

PubMed

Ellison, Aaron M; Parker, Jerelyn N

2002-06-01

Plant ecologists continue to grapple with Reid's paradox, the observation that dispersal distances of most herbs and trees are too limited to account for their recolonization of northern latitudes following glacial recession. As global climate changes and natural habitats become increasingly fragmented, understanding patterns of seed dispersal and the potential for long-distance colonization takes on new importance. We studied the dispersal and establishment of the northern pitcher plant Sarracenia purpurea, which grows commonly in isolated bogs throughout Canada and eastern North America. Median dispersal distance of S. purpurea is only 5 cm, which is insufficient to explain its occurrence throughout formerly glaciated regions of North America. Establishment probability of seeds in the field is approximately 5%, and juveniles are normally found clustered around adult plants. The large-scale population genetic structure of this species can be accounted for by rare long-distance dispersal events, but its predictable occurrence in isolated habitats requires additional explanation. Reid's paradox remains an open question, and predicting long-range colonization into fragmented habitats by species with limited dispersal ability is a novel challenge.

Identification of Echinococcus granulosus strains using polymerase chain reaction-restriction fragment length polymorphism amongst livestock in Moroto district, Uganda.

PubMed

Chamai, Martin; Omadang, Leonard; Erume, Joseph; Ocaido, Michael; Oba, Peter; Othieno, Emmanuel; Bonaventure, Straton; Kitibwa, Annah

2016-07-29

A descriptive study was conducted to identify the different strains of Echinococcus granulosus occurring in livestock in Moroto district, Uganda. Echinococcus cysts from 104 domestic animals, including cattle, sheep, goats and camels, were taken and examined by microscopy, polymerase chain reaction with restriction fragment length polymorphism and Sanger DNA sequencing. Echinococcus granulosus genotypes or strains were identified through use of Bioinformatics tools: BioEdit, BLAST and MEGA6. The major finding of this study was the existence of a limited number of E. granulosus genotypes from cattle, goats, sheep and camels. The most predominant genotype was G1 (96.05%), corresponding to the common sheep strain. To a limited extent (3.95%), the study revealed the existence of Echinococcus canadensis G6/7 in three (n = 3) of the E. granulosus-positive samples. No other strains of E. granulosus were identified. It was concluded that the common sheep strain of Echinococcus sensu stricto and G6/7 of E. canadensis were responsible for echinococcal disease in Moroto district, Uganda.

Nicked apomyoglobin: a noncovalent complex of two polypeptide fragments comprising the entire protein chain.

PubMed

Musi, Valeria; Spolaore, Barbara; Picotti, Paola; Zambonin, Marcello; De Filippis, Vincenzo; Fontana, Angelo

2004-05-25

Limited proteolysis of the 153-residue chain of horse apomyoglobin (apoMb) by thermolysin results in the selective cleavage of the peptide bond Pro88-Leu89. The N-terminal (residues 1-88) and C-terminal (residues 89-153) fragments of apoMb were isolated to homogeneity and their conformational and association properties investigated in detail. Far-UV circular dichroism (CD) measurements revealed that both fragments in isolation acquire a high content of helical secondary structure, while near-UV CD indicated the absence of tertiary structure. A 1:1 mixture of the fragments leads to a tight noncovalent protein complex (1-88/89-153, nicked apoMb), characterized by secondary and tertiary structures similar to those of intact apoMb. The apoMb complex binds heme in a nativelike manner, as given by CD measurements in the Soret region. Second-derivative absorption spectra in the 250-300 nm region provided evidence that the degree of exposure of Tyr residues in the nicked species is similar to that of the intact protein at neutral pH. Also, the microenvironment of Trp residues, located in positions 7 and 14 of the 153-residue chain of the protein, is similar in both protein species, as given by fluorescence emission data. Moreover, in analogy to intact apoMb, the nicked protein binds the hydrophobic dye 1-anilinonaphthalene-8-sulfonate (ANS). Taken together, our results indicate that the two proteolytic fragments 1-88 and 89-153 of apoMb adopt partly folded states characterized by sufficiently nativelike conformational features that promote their specific association and mutual stabilization into a nicked protein species much resembling in its structural features intact apoMb. It is suggested that the formation of a noncovalent complex upon fragment complementation can mimic the protein folding process of the entire protein chain, with the difference that the folding of the complementary fragments is an intermolecular process. In particular, this study emphasizes the importance of interactions between marginally stable elements of secondary structure in promoting the tertiary contacts of a native protein. Considering that apoMb has been extensively used as a paradigm in protein folding studies for the past few decades, the novel fragment complementing system of apoMb here described appears to be very useful for investigating the initial as well as late events in protein folding.

Is health care financing in Uganda equitable?

PubMed

Zikusooka, C M; Kyomuhang, R; Orem, J N; Tumwine, M

2009-10-01

Health care financing provides the resources and economic incentives for operating health systems and is a key determinant of health system performance. Equitable financing is based on: financial protection, progressive financing and cross-subsidies. This paper describes Uganda's health care financing landscape and documents the key equity issues associated with the current financing mechanisms. We extensively reviewed government documents and relevant literature and conducted key informant interviews, with the aim of assessing whether Uganda's health care financing mechanisms exhibited the key principles of fair financing. Uganda's health sector remains significantly under-funded, mainly relying on private sources of financing, especially out-of-pocket spending. At 9.6 % of total government expenditure, public spending on health is far below the Abuja target of 15% that GoU committed to. Prepayments form a small proportion of funding for Uganda's health sector. There is limited cross-subsidisation and high fragmentation within and between health financing mechanisms, mainly due to high reliance on out-of-pocket payments and limited prepayment mechanisms. Without compulsory health insurance and low coverage of private health insurance, Uganda has limited pooling of resources, and hence minimal cross-subsidisation. Although tax revenue is equitable, the remaining financing mechanisms for Uganda are inequitable due to their regressive nature, their lack of financial protection and limited cross-subsidisation. Overall, Uganda's current health financing is inequitable and fragmented. The government should take explicit action to promote equitable health care financing by establishing pre-payment schemes, enhancing cross-subsidisation mechanisms and through appropriate integration of financing mechanisms.

Role of the Cationic C-Terminal Segment of Melittin on Membrane Fragmentation.

PubMed

Therrien, Alexandre; Fournier, Alain; Lafleur, Michel

2016-05-05

The widespread distribution of cationic antimicrobial peptides capable of membrane fragmentation in nature underlines their importance to living organisms. In the present work, we determined the impact of the electrostatic interactions associated with the cationic C-terminal segment of melittin, a 26-amino acid peptide from bee venom (net charge +6), on its binding to model membranes and on the resulting fragmentation. In order to detail the role played by the C-terminal charges, we prepared a melittin analogue for which the four cationic amino acids in positions 21-24 were substituted with the polar residue citrulline, providing a peptide with the same length and amphiphilicity but with a lower net charge (+2). We compared the peptide bilayer affinity and the membrane fragmentation for bilayers prepared from 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)/1,2-dipalmitoyl-sn-glycero-3-phospho-l-serine (DPPS) mixtures. It is shown that neutralization of the C-terminal considerably increased melittin affinity for zwitterionic membranes. The unfavorable contribution associated with transferring the cationic C-terminal in a less polar environment was reduced, leaving the hydrophobic interactions, which drive the peptide insertion in bilayers, with limited counterbalancing interactions. The presence of negatively charged lipids (DPPS) in bilayers increased melittin binding by introducing attractive electrostatic interactions, the augmentation being, as expected, greater for native melittin than for its citrullinated analogue. The membrane fragmentation power of the peptide was shown to be controlled by electrostatic interactions and could be modulated by the charge carried by both the membrane and the lytic peptide. The analysis of the lipid composition of the extracted fragments from DPPC/DPPS bilayers revealed no lipid specificity. It is proposed that extended phase separations are more susceptible to lead to the extraction of a lipid species in a specific manner than a specific lipid-peptide affinity. The present work on the lipid extraction by melittin and citrullinated melittin with model membranes emphasizes the complex relation between the affinity, the lipid extraction/membrane fragmentation, and the lipid specificity.

Introgression of a 4D chromosomal fragment into durum wheat confers aluminium tolerance

PubMed Central

Han, Chang; Ryan, Peter R.; Yan, ZeHong; Delhaize, Emmanuel

2014-01-01

Background and Aim Aluminium (Al3+) inhibits root growth of sensitive plant species and is a key factor that limits durum wheat (Triticum turgidum) production on acid soils. The aim of this study was to enhance the Al3+ tolerance of an elite durum cultivar by introgression of a chromosomal fragment from hexaploid wheat (Triticum aestivum) that possesses an Al3+ tolerance gene. Methods A 4D(4B) substitution line of durum wheat ‘Langdon’ was backcrossed to ‘Jandaroi’, a current semi-dwarf Australian durum. In the second backcross, using ‘Jandaroi’ as the recurrent parent, a seedling was identified where TaALMT1 on chromosome 4D was recombined with the Rht-B1b locus on chromosome 4B to yield an Al3+-tolerant seedling with a semi-dwarf habit. This seedling was used in a third backcross to generate homozygous sister lines with contrasting Al3+ tolerances. The backcrossed lines were characterized and compared with selected cultivars of hexaploid wheat for their Al3+ and Na+ tolerances in hydroponic culture as well as in short-term experiments to assess their growth on acid soil. Key Results Analysis of sister lines derived from the third backcross showed that the 4D chromosomal fragment substantially enhanced Al3+ tolerance. The ability to exclude Na+ from leaves was also enhanced, indicating that the chromosomal fragment possessed the Kna1 salt tolerance locus. Although Al3+ tolerance of seminal roots was enhanced in acid soil, the development of fine roots was not as robust as found in Al3+-tolerant lines of hexaploid wheat. Analysis of plant characteristics in the absence of Al3+ toxicity showed that the introgressed fragment did not affect total grain yield but reduced the weight of individual grains. Conclusions The results show that it is possible to increase substantially the Al3+ tolerance of an elite durum wheat cultivar by introgression of a 4D chromosomal fragment. Further improvements are possible, such as introducing additional genes to enhance the Al3+ tolerance of fine roots and by eliminating the locus on the chromosomal fragment responsible for smaller grain weights. PMID:24737716

Calcification rate and the stable carbon, oxygen, and nitrogen isotopes in the skeleton, host tissue, and zooxanthellae of bleached and recovering Hawaiian corals

NASA Astrophysics Data System (ADS)

Rodrigues, Lisa J.; Grottoli, Andréa G.

2006-06-01

We tested the effectiveness of stable isotopes as recorders of physiological changes that occur during coral bleaching and recovery. Montipora capitata and Porites compressa fragments were bleached in outdoor tanks with seawater temperature raised to 30 °C (treatment corals) for one month. Additional fragments were maintained at 27 °C in separate tanks (control corals). After one month, (0 months recovery), buoyant weight was measured and a subset of fragments was frozen. Remaining fragments were returned to the reef for recovery. After 1.5, 4, and 8 months, fragments were collected, measured for buoyant weight, and frozen. Fragments were analyzed for stable carbon and oxygen isotopic compositions of the skeleton (δ 13C s; δ 18O s) and nitrogen and carbon isotopic compositions of the host tissue (δ 15N h; δ 13C h) and zooxanthellae (δ 15N z; δ 13C z). δ 13C s decreased immediately after bleaching in M. capitata, but not in P. compressa. δ 18O s of both species failed to record the warming event. During the remaining months of recovery, δ 13C s and δ 18O s were more enriched in treatment than control corals due to decreases in calcification and metabolic fractionation during that time. Increased δ 15N h of treatment P. compressa may be due to expelled zooxanthellae during bleaching and recovery. Increased δ 15N z at 1.5 months in treatment fragments of both species reflects the increased incorporation of dissolved inorganic nitrogen to facilitate mitotic cell division and/or chl a/cell recovery. Changes in δ 13C h and δ 13C z at 1.5 months in treatment M. capitata indicated a large increase in heterotrophically acquired carbon relative to photosynthetically fixed carbon. We experimentally show that isotopes in coral skeleton, host tissue and zooxanthellae can be used to verify physiological changes during bleaching and recovery, but their use as a proxy for past bleaching events in the skeletal record is limited.

Study of the family of a patient with male-limited precocious puberty (MPP) due to T1193C transition in exon 11 of LH receptor gene.

PubMed

Ignacak, M; Starzyk, J; Dziatkowiak, H; Trzeciak, W H

2002-03-01

Molecular diagnostics of the LHR gene was conducted in a 5-year-old boy with clinical symptoms and hormonal profile typical of precocious puberty. His parents and 4 sisters were also diagnosed. Single-strand conformation polymorphism analysis under temperature gradient conditions (Multitemperature SSCP) of 3 overlapping fragments of exon 11 of LHR gene revealed a mutation in the fragment spanning nucleotides 1072 to 1804. This mutation was found in the patient, in his mother and in his 4 sisters, and was confirmed by digestion with the use of restriction enzyme Bbr Cl. Direct sequencing revealed a heterozygous T1193C transition in the DNA fragment of the patient and in one of the alleles of his mother's and sister's DNA. This mutation causes Met398Thr substitution in the second transmembrane helix and results in a constitutive activation of LH receptor. This is the second identical mutation detected in Poland and one of the 7 identified so far in the world population.

Accurate phylogenetic classification of DNA fragments based onsequence composition

DOE Office of Scientific and Technical Information (OSTI.GOV)

McHardy, Alice C.; Garcia Martin, Hector; Tsirigos, Aristotelis

2006-05-01

Metagenome studies have retrieved vast amounts of sequenceout of a variety of environments, leading to novel discoveries and greatinsights into the uncultured microbial world. Except for very simplecommunities, diversity makes sequence assembly and analysis a verychallenging problem. To understand the structure a 5 nd function ofmicrobial communities, a taxonomic characterization of the obtainedsequence fragments is highly desirable, yet currently limited mostly tothose sequences that contain phylogenetic marker genes. We show that forclades at the rank of domain down to genus, sequence composition allowsthe very accurate phylogenetic 10 characterization of genomic sequence.We developed a composition-based classifier, PhyloPythia, for de novophylogenetic sequencemore » characterization and have trained it on adata setof 340 genomes. By extensive evaluation experiments we show that themethodis accurate across all taxonomic ranks considered, even forsequences that originate fromnovel organisms and are as short as 1kb.Application to two metagenome datasets 15 obtained from samples ofphosphorus-removing sludge showed that the method allows the accurateclassification at genus level of most sequence fragments from thedominant populations, while at the same time correctly characterizingeven larger parts of the samples at higher taxonomic levels.« less

Increase in the astaxanthin synthase gene (crtS) dose by in vivo DNA fragment assembly in Xanthophyllomyces dendrorhous

PubMed Central

2013-01-01

Background Xanthophyllomyces dendrorhous is a basidiomycetous yeast that is relevant to biotechnology, as it can synthesize the carotenoid astaxanthin. However, the astaxanthin levels produced by wild-type strains are low. Although different approaches for promoting increased astaxanthin production have been attempted, no commercially competitive results have been obtained thus far. A promising alternative to facilitate the production of carotenoids in this yeast involves the use of genetic modification. However, a major limitation is the few available molecular tools to manipulate X. dendrorhous. Results In this work, the DNA assembler methodology that was previously described in Saccharomyces cerevisiae was successfully applied to assemble DNA fragments in vivo and integrate these fragments into the genome of X. dendrorhous by homologous recombination in only one transformation event. Using this method, the gene encoding astaxanthin synthase (crtS) was overexpressed in X. dendrorhous and a higher level of astaxanthin was produced. Conclusions This methodology could be used to easily and rapidly overexpress individual genes or combinations of genes simultaneously in X. dendrorhous, eliminating numerous steps involved in conventional cloning methods. PMID:24103677

Enzyme-linked small-molecule detection using split aptamer ligation.

PubMed

Sharma, Ashwani K; Kent, Alexandra D; Heemstra, Jennifer M

2012-07-17

Here we report an aptamer-based analogue of the widely used sandwich enzyme-linked immunosorbent assay (ELISA). This assay utilizes the cocaine split aptamer, which is comprised of two DNA strands that only assemble in the presence of the target small molecule. One split aptamer fragment is immobilized on a microplate, then a test sample is added containing the second split aptamer fragment. If cocaine is present in the test sample, it directs assembly of the split aptamer and promotes a chemical ligation between azide and cyclooctyne functional groups appended to the termini of the split aptamer fragments. Ligation results in covalent attachment of biotin to the microplate and provides a colorimetric output upon conjugation to streptavidin-horseradish peroxidase. Using this assay, we demonstrate detection of cocaine at concentrations of 100 nM-100 μM in buffer and 1-100 μM human blood serum. The detection limit of 1 μM in serum represents an improvement of two orders of magnitude over previously reported split aptamer-based sensors and highlights the utility of covalently trapping split aptamer assembly events.

The movement ecology and dynamics of plant communities in fragmented landscapes.

PubMed

Damschen, Ellen I; Brudvig, Lars A; Haddad, Nick M; Levey, Douglas J; Orrock, John L; Tewksbury, Joshua J

2008-12-09

A conceptual model of movement ecology has recently been advanced to explain all movement by considering the interaction of four elements: internal state, motion capacity, navigation capacities, and external factors. We modified this framework to generate predictions for species richness dynamics of fragmented plant communities and tested them in experimental landscapes across a 7-year time series. We found that two external factors, dispersal vectors and habitat features, affected species colonization and recolonization in habitat fragments and their effects varied and depended on motion capacity. Bird-dispersed species richness showed connectivity effects that reached an asymptote over time, but no edge effects, whereas wind-dispersed species richness showed steadily accumulating edge and connectivity effects, with no indication of an asymptote. Unassisted species also showed increasing differences caused by connectivity over time, whereas edges had no effect. Our limited use of proxies for movement ecology (e.g., dispersal mode as a proxy for motion capacity) resulted in moderate predictive power for communities and, in some cases, highlighted the importance of a more complete understanding of movement ecology for predicting how landscape conservation actions affect plant community dynamics.

Different pollinator assemblages ensure reproductive success of Cleisostoma linearilobatum (Orchidaceae) in fragmented holy hill forest and traditional tea garden.

PubMed

Zhou, Xiang; Liu, Qiang; Han, Jessie Yc; Gao, JiangYun

2016-02-24

Orchids are generally recognized to have specialist pollination systems and low fruit set is often thought to be characteristic of the family. In this study, we investigated the reproductive ecology of Cleisostoma linearilobatum, an epiphytic tropical orchid, in a holy hill forest fragment and a traditional tea garden in SW China using comparable methods. C. linearilobatum is self-compatible and dependent on insects for pollination. Fruit production in natural conditions was both pollinator- and resource-limited. However, the natural fruit set remained stable over multiple years at both sites. Pollination observations showed that C. linearilobatum has a generalized pollination system and seven insect species were observed as legitimate pollinators. Although the visit frequencies of different pollinators were different in the two sites, the pollinator assemblages ensured reproductive success of C. linearilobatum in both study sites over multiple years. The results partly explain why C. linearilobatum is so successful in the area, and also suggest that holy hill forest fragments and traditional tea gardens in Xishuangbanna are important in preserving orchids, especially those with generalist pollination.

Different pollinator assemblages ensure reproductive success of Cleisostoma linearilobatum (Orchidaceae) in fragmented holy hill forest and traditional tea garden

PubMed Central

Zhou, Xiang; Liu, Qiang; Han, Jessie Yc; Gao, JiangYun

2016-01-01

Orchids are generally recognized to have specialist pollination systems and low fruit set is often thought to be characteristic of the family. In this study, we investigated the reproductive ecology of Cleisostoma linearilobatum, an epiphytic tropical orchid, in a holy hill forest fragment and a traditional tea garden in SW China using comparable methods. C. linearilobatum is self-compatible and dependent on insects for pollination. Fruit production in natural conditions was both pollinator- and resource-limited. However, the natural fruit set remained stable over multiple years at both sites. Pollination observations showed that C. linearilobatum has a generalized pollination system and seven insect species were observed as legitimate pollinators. Although the visit frequencies of different pollinators were different in the two sites, the pollinator assemblages ensured reproductive success of C. linearilobatum in both study sites over multiple years. The results partly explain why C. linearilobatum is so successful in the area, and also suggest that holy hill forest fragments and traditional tea gardens in Xishuangbanna are important in preserving orchids, especially those with generalist pollination. PMID:26907369

Quantitative spatiotemporal analysis of antibody fragment diffusion and endocytic consumption in tumor spheroids.

PubMed

Thurber, Greg M; Wittrup, K Dane

2008-05-01

Antibody-based cancer treatment depends upon distribution of the targeting macromolecule throughout tumor tissue, and spatial heterogeneity could significantly limit efficacy in many cases. Antibody distribution in tumor tissue is a function of drug dosage, antigen concentration, binding affinity, antigen internalization, drug extravasation from blood vessels, diffusion in the tumor extracellular matrix, and systemic clearance rates. We have isolated the effects of a subset of these variables by live-cell microscopic imaging of single-chain antibody fragments against carcinoembryonic antigen in LS174T tumor spheroids. The measured rates of scFv penetration and retention were compared with theoretical predictions based on simple scaling criteria. The theory predicts that antibody dose must be large enough to drive a sufficient diffusive flux of antibody to overcome cellular internalization, and exposure time must be long enough to allow penetration to the spheroid center. The experimental results in spheroids are quantitatively consistent with these predictions. Therefore, simple scaling criteria can be applied to accurately predict antibody and antibody fragment penetration distance in tumor tissue.

Quantitative Spatiotemporal Analysis of Antibody Fragment Diffusion and Endocytic Consumption in Tumor Spheroids

PubMed Central

Thurber, Greg M.; Wittrup, K. Dane

2010-01-01

Antibody-based cancer treatment depends upon distribution of the targeting macromolecule throughout tumor tissue, and spatial heterogeneity could significantly limit efficacy in many cases. Antibody distribution in tumor tissue is a function of drug dosage, antigen concentration, binding affinity, antigen internalization, drug extravasation from blood vessels, diffusion in the tumor extracellular matrix, and systemic clearance rates. We have isolated the effects of a subset of these variables by live-cell microscopic imaging of single-chain antibody fragments against carcinoembryonic antigen in LS174T tumor spheroids. The measured rates of scFv penetration and retention were compared with theoretical predictions based on simple scaling criteria. The theory predicts that antibody dose must be large enough to drive a sufficient diffusive flux of antibody to overcome cellular internalization, and exposure time must be long enough to allow penetration to the spheroid center. The experimental results in spheroids are quantitatively consistent with these predictions. Therefore, simple scaling criteria can be applied to accurately predict antibody and antibody fragment penetration distance in tumor tissue. PMID:18451160

Site-specific PEGylation of an anti-CEA/CD3 bispecific antibody improves its antitumor efficacy

PubMed Central

Pan, Haitao; Liu, Jiayu; Deng, Wentong; Xing, Jieyu; Li, Qing; Wang, Zhong

2018-01-01

Introduction Bispecific antibodies that engage immune cells to kill cancer cells are actively pursued in cancer immunotherapy. Different types of bispecific antibodies, including single-chain fragments, Fab fragments, nanobodies, and immunoglobulin Gs (IgGs), have been studied. However, the low molecular weight of bispecific antibodies with single-chain or Fab fragments generally leads to their rapid clearance in vivo, which limits the therapeutic potential of these bispecific antibodies. Materials and methods In this study, we used a site-specific PEGylation strategy to modify the bispecific single-domain antibody-linked Fab (S-Fab), which was designed by linking an anticarcinoembryonic antigen (anti-CEA) nanobody with an anti-CD3 Fab. Results The half-life (t1/2) of PEGylated S-Fab (polyethylene glycol-S-Fab) was increased 12-fold in vivo with a slightly decreased tumor cell cytotoxicity in vitro as well as more potent tumor growth inhibition in vivo compared to S-Fab. Conclusion This study demonstrated that PEGylation is an effective approach to enhance the antitumor efficacy of bispecific antibodies. PMID:29881272

Spatial Ecology of Estuarine Crocodile (Crocodylus porosus) Nesting in a Fragmented Landscape.

PubMed

Evans, Luke J; Jones, T Hefin; Pang, Keeyen; Saimin, Silvester; Goossens, Benoit

2016-09-19

The role that oil palm plays in the Lower Kinabatangan region of Eastern Sabah is of considerable scientific and conservation interest, providing a model habitat for many tropical regions as they become increasingly fragmented. Crocodilians, as apex predators, widely distributed throughout the tropics, are ideal indicator species for ecosystem health. Drones (or unmanned aerial vehicles (UAVs)) were used to identify crocodile nests in a fragmented landscape. Flights were targeted through the use of fuzzy overlay models and nests located primarily in areas indicated as suitable habitat. Nests displayed a number of similarities in terms of habitat characteristics allowing for refined modelling of survey locations. As well as being more cost-effective compared to traditional methods of nesting survey, the use of drones also enabled a larger survey area to be completed albeit with a limited number of flights. The study provides a methodology for targeted nest surveying, as well as a low-cost repeatable flight methodology. This approach has potential for widespread applicability across a range of species and for a variety of study designs.

Spatial Ecology of Estuarine Crocodile (Crocodylus porosus) Nesting in a Fragmented Landscape

PubMed Central

Evans, Luke J.; Jones, T. Hefin; Pang, Keeyen; Saimin, Silvester; Goossens, Benoit

2016-01-01

The role that oil palm plays in the Lower Kinabatangan region of Eastern Sabah is of considerable scientific and conservation interest, providing a model habitat for many tropical regions as they become increasingly fragmented. Crocodilians, as apex predators, widely distributed throughout the tropics, are ideal indicator species for ecosystem health. Drones (or unmanned aerial vehicles (UAVs)) were used to identify crocodile nests in a fragmented landscape. Flights were targeted through the use of fuzzy overlay models and nests located primarily in areas indicated as suitable habitat. Nests displayed a number of similarities in terms of habitat characteristics allowing for refined modelling of survey locations. As well as being more cost-effective compared to traditional methods of nesting survey, the use of drones also enabled a larger survey area to be completed albeit with a limited number of flights. The study provides a methodology for targeted nest surveying, as well as a low-cost repeatable flight methodology. This approach has potential for widespread applicability across a range of species and for a variety of study designs. PMID:27657065

Organomineral interactions as an important mechanism for stabilisation of bacterial residues in soil

NASA Astrophysics Data System (ADS)

Miltner, Anja; Achtenhagen, Jan; Kästner, Matthias

2017-04-01

Although plant material is the original input of organic matter to soils, microbial residues have been identified to contribute to a large extent to soil organic matter. However, until now it is unclear how microbial residues are stabilised in soil and protected from degradation. We hypothesised that organomineral interactions, in particular encrustation by oxides, may play an important role, which might vary depending on environmental conditions, e.g. redox potential. Therefore we produced 14C-labelled Escherichia coli cells and cell envelope fragments and coprecipitated these materials with Fe oxide or Al oxide. Mineral-free (control) and mineral-encrusted bacterial residues were incubated for 345 days at 20˚ C under either oxic or oxygen-limited conditions, and mineralisation was quantified by scintillation counting of the CO2 produced during incubation. Oxygen limitation was achieved by first exchanging the atmosphere in the incubation vessels with dinitrogen gas. After 100 days of incubation, the anoxic treatments were waterlogged to further decrease the redox potential, and after 290 days, glucose and nutrients were supplied to all treatments in order to foster microbial activity and consumption of electron acceptors. The mineralisation curves were fitted by double-exponential (0-100 days), first-order kinetic (100-290 days) and linear (290-345 days) models. The model parameters were tested for significant differences between the treatments by three-way ANOVA with post-hoc Bonferroni t-test. We found that encrustation by the oxides significantly reduced mineralisation of the bacterial residues. This effect was inversed by reductive dissolution of Fe oxides after substrate and nutrient addition to the oxygen-limited treatments, suggesting a significant role of the encrustation in stabilisation of the bacterial residues. We also observed that bacterial cell envelope fragments were generally slightly more resistant to mineralisation than whole cells. The results indicate that bacterial residues, in particular cell envelope fragments, may be stabilised in soil by organomineral interactions as long as the minerals are stable in soil.

A dated phylogeny and collection records reveal repeated biome shifts in the African genus Coccinia (Cucurbitaceae).

PubMed

Holstein, Norbert; Renner, Susanne S

2011-01-26

Conservatism in climatic tolerance may limit geographic range expansion and should enhance the effects of habitat fragmentation on population subdivision. Here we study the effects of historical climate change, and the associated habitat fragmentation, on diversification in the mostly sub-Saharan cucurbit genus Coccinia, which has 27 species in a broad range of biota from semi-arid habitats to mist forests. Species limits were inferred from morphology, and nuclear and plastid DNA sequence data, using multiple individuals for the widespread species. Climatic tolerances were assessed from the occurrences of 1189 geo-referenced collections and WorldClim variables. Nuclear and plastid gene trees included 35 or 65 accessions, representing up to 25 species. The data revealed four species groups, one in southern Africa, one in Central and West African rain forest, one widespread but absent from Central and West African rain forest, and one that occurs from East Africa to southern Africa. A few individuals are differently placed in the plastid and nuclear (LFY) trees or contain two ITS sequence types, indicating hybridization. A molecular clock suggests that the diversification of Coccinia began about 6.9 Ma ago, with most of the extant species diversity dating to the Pliocene. Ancestral biome reconstruction reveals six switches between semi-arid habitats, woodland, and forest, and members of several species pairs differ significantly in their tolerance of different precipitation regimes. The most surprising findings of this study are the frequent biome shifts (in a relatively small clade) over just 6 - 7 million years and the limited diversification during and since the Pleistocene. Pleistocene climate oscillations may have been too rapid or too shallow for full reproductive barriers to develop among fragmented populations of Coccinia, which would explain the apparently still ongoing hybridization between certain species. Steeper ecological gradients in East Africa and South Africa appear to have resulted in more advanced allopatric speciation there.

Limited mate availability decreases reproductive success of fragmented populations of Linnaea borealis, a rare, clonal self-incompatible plant

PubMed Central

Scobie, A. R.; Wilcock, C. C.

2009-01-01

Background and Aims Small populations of rare plant species are increasingly reported to have high levels of reproductive failure. The objective of this study was to understand the principal constraints on sexual reproduction in small fragmented populations of a rare clonal self-incompatible plant. Methods The pollinator spectrum, diversity of flower colour, natural pollination and fruit-set levels of L. borealis were examined in Scotland. Artificially crossed seed production was compared within and between different flower colour types and patches. Key Results Linnaea borealis was pollinated by a diverse spectrum of insect species and the principal pollinators were muscid, syrphid and empid flies which mostly moved only small distances (<0·25 m) between flowers when foraging. Natural pollination levels were high, indicating high pollinator effectiveness, but fruit set was very low in most patches. Flower colour diversity was low in most patches and only those with a diversity of flower colour types had high fruiting success. Pollination experiments showed L. borealis to be highly self-incompatible and artificial crosses within and between patches and flower colour types confirmed that low fruit success was the result of a lack of compatible mates and limited pollen movement between them. Evidence of isolation from pollen exchange was apparent at as little as 6 m and severe at 30 m and beyond. Conclusions Limited mate availability and isolation from pollen exchange compromise the reproductive success of fragmented populations of L. borealis in Scotland. A diversity of compatible mates situated within close proximity (<6 m) is the key requirement to ensure high natural fruiting success. This study emphasizes that an understanding of the breeding system, pollinator spectrum and potential for interconnectivity via pollinator movement are fundamental to identify isolation distances and to establish when conservation intervention is necessary for rare species. PMID:19181748

Development of one novel multiple-target plasmid for duplex quantitative PCR analysis of roundup ready soybean.

PubMed

Zhang, Haibo; Yang, Litao; Guo, Jinchao; Li, Xiang; Jiang, Lingxi; Zhang, Dabing

2008-07-23

To enforce the labeling regulations of genetically modified organisms (GMOs), the application of reference molecules as calibrators is becoming essential for practical quantification of GMOs. However, the reported reference molecules with tandem marker multiple targets have been proved not suitable for duplex PCR analysis. In this study, we developed one unique plasmid molecule based on one pMD-18T vector with three exogenous target DNA fragments of Roundup Ready soybean GTS 40-3-2 (RRS), that is, CaMV35S, NOS, and RRS event fragments, plus one fragment of soybean endogenous Lectin gene. This Lectin gene fragment was separated from the three exogenous target DNA fragments of RRS by inserting one 2.6 kb DNA fragment with no relatedness to RRS detection targets in this resultant plasmid. Then, we proved that this design allows the quantification of RRS using the three duplex real-time PCR assays targeting CaMV35S, NOS, and RRS events employing this reference molecule as the calibrator. In these duplex PCR assays, the limits of detection (LOD) and quantification (LOQ) were 10 and 50 copies, respectively. For the quantitative analysis of practical RRS samples, the results of accuracy and precision were similar to those of simplex PCR assays, for instance, the quantitative results were at the 1% level, the mean bias of the simplex and duplex PCR were 4.0% and 4.6%, respectively, and the statistic analysis ( t-test) showed that the quantitative data from duplex and simplex PCR had no significant discrepancy for each soybean sample. Obviously, duplex PCR analysis has the advantages of saving the costs of PCR reaction and reducing the experimental errors in simplex PCR testing. The strategy reported in the present study will be helpful for the development of new reference molecules suitable for duplex PCR quantitative assays of GMOs.

Flux-based Enrichment Ratios of Throughfall and Stemflow Found to Vary Significantly within Urban Fragments and Along an Urban-to-Rural Gradient

NASA Astrophysics Data System (ADS)

Dowtin, A. L.; Levia, D. F., Jr.

2017-12-01

Throughfall and stemflow are important inputs of water and solutes to forest soils in both rural and urban forests. In metropolitan wooded ecosystems, a number of factors can affect flux-based enrichment ratios, including combustion of fossil fuels and proximity to industry. Use of flux-based enrichment ratios provides a means by which this modification of net precipitation chemistry can be quantified for both throughfall and stemflow, and allows for a characterization of the relative contributions of stemflow and throughfall in the delivery of nutrients and pollutants to forest soils. This study utilizes five mixed deciduous forest stands along an urban-to-rural gradient (3 urban fragments, 1 suburban fragment, and a portion of 1 contiguous rural forest) within a medium-sized metropolitan region of the United States' Northeast megalopolis, to determine how the size, shape, structure, and geographic context of remnant forest fragments determine hydrologic and solute fluxes within them. In situ observations of throughfall and stemflow (the latter of which is limited to Quercus rubra and Quercus alba) within each study plot allow for an identification and characterization of the spatial variability in solute fluxes within and between the respective sites. Preliminary observations indicate significant intra-site variability in solute concentrations as observed in both throughfall and stemflow, with higher concentrations along the respective windward edges of the study plots than at greater depths into their interiors. Higher flux-based stemflow enrichment ratios, for both Q. rubra and Q. alba, were also evident for certain ions (i.e., S2-, NO3-) in the urban forest fragments, with significantly lower ratios observed at the suburban and rural sites. Findings from this research are intended to aid in quantifying the spatial variability of the hydrologic and hydrochemical ecosystem service provisions of remnant metropolitan forest fragments. This research is supported in part by National Science Foundation grant Reference Number BCS-1459116.

The fragmented testis method: development and its advantages of a new quantitative evaluation technique for detection of testis-ova in male fish.

PubMed

Lin, Bin-Le; Hagino, Satoshi; Kagoshima, Michio; Iwamatsu, Takashi

2009-02-01

A new quantitative evaluation technique, termed the fragmented testis method, has been developed for the detection of testis-ova in genotypic male fish using the medaka (Oryzias latipes). The routine traditional histological method for detection of testis-ova in male fish exposed to estrogens or suspected endocrine-disrupting chemicals has several disadvantages, including possible oversight of testis-ova due to limited sampling of selected tissue sections. The method we have developed here allows for the accurate determination of the developmental stages and the number and the size of testis-ova in a whole testis. Each testis was removed from the fish specimen, fixed with 10% buffered formalin solution, and then divided into small fragments on a glass slide with a dissecting needle or scalpel and aciform forceps in glycerin solution containing a small amount of methylene blue or toluidine blue. If present, all developing testis-ova of various sizes in fragmented testicular tissues were clearly stained and were observable under a dissecting microscope. Testis-ova occurred in controls were ascertained, while spermatozoa were also distinguishable using this method. This proved to be a convenient and cost-effective method for quantitatively evaluating testis-ova appearance in fish, and it may help to clarify the mechanism of testis-ova formation and the biological significance of testis-ova in future studies of endocrine disruption.

Sulfolobus chromatin proteins modulate strand displacement by DNA polymerase B1

PubMed Central

Sun, Fei; Huang, Li

2013-01-01

Strand displacement by a DNA polymerase serves a key role in Okazaki fragment maturation, which involves displacement of the RNA primer of the preexisting Okazaki fragment into a flap structure, and subsequent flap removal and fragment ligation. We investigated the role of Sulfolobus chromatin proteins Sso7d and Cren7 in strand displacement by DNA polymerase B1 (PolB1) from the hyperthermophilic archaeon Sulfolobus solfataricus. PolB1 showed a robust strand displacement activity and was capable of synthesizing thousands of nucleotides on a DNA-primed 72-nt single-stranded circular DNA template. This activity was inhibited by both Sso7d and Cren7, which limited the flap length to 3–4 nt at saturating concentrations. However, neither protein inhibited RNA displacement on an RNA-primed single-stranded DNA minicircle by PolB1. Strand displacement remained sensitive to modulation by the chromatin proteins when PolB1 was in association with proliferating cell nuclear antigen. Inhibition of DNA instead of RNA strand displacement by the chromatin proteins is consistent with the finding that double-stranded DNA was more efficiently bound and stabilized than an RNA:DNA duplex by these proteins. Our results suggest that Sulfolobus chromatin proteins modulate strand displacement by PolB1, permitting efficient removal of the RNA primer while inhibiting excessive displacement of the newly synthesized DNA strand during Okazaki fragment maturation. PMID:23821667

Multiplex Real-Time PCR Assays that Measure the Abundance of Extremely Rare Mutations Associated with Cancer

PubMed Central

Vargas, Diana Y.; Kramer, Fred Russell; Tyagi, Sanjay; Marras, Salvatore A. E.

2016-01-01

We describe the use of “SuperSelective” primers that enable the detection and quantitation of somatic mutations whose presence relates to cancer diagnosis, prognosis, and therapy, in real-time PCR assays that can potentially analyze rare DNA fragments present in blood samples (liquid biopsies). The design of these deoxyribonucleotide primers incorporates both a relatively long “5' anchor sequence” that hybridizes strongly to target DNA fragments, and a very short, physically and functionally separate, “3' foot sequence” that is perfectly complementary to the mutant target sequence, but mismatches the wild-type sequence. As few as ten mutant fragments can reliably be detected in the presence of 1,000,000 wild-type fragments, even when the difference between the mutant and the wild type is only a single nucleotide polymorphism. Multiplex PCR assays employing a set of SuperSelective primers, and a corresponding set of differently colored molecular beacon probes, can be used in situations where the different mutations, though occurring in different cells, are located in the same codon. These non-symmetric real-time multiplex PCR assays contain limited concentrations of each SuperSelective primer, thereby enabling the simultaneous determination of each mutation’s abundance by comparing its threshold value to the threshold value of a reference gene present in the sample. PMID:27244445

Studies of Dynamic Protein-Protein Interactions in Bacteria Using Renilla Luciferase Complementation Are Undermined by Nonspecific Enzyme Inhibition

PubMed Central

Hatzios, Stavroula K.; Ringgaard, Simon; Davis, Brigid M.; Waldor, Matthew K.

2012-01-01

The luciferase protein fragment complementation assay is a powerful tool for studying protein-protein interactions. Two inactive fragments of luciferase are genetically fused to interacting proteins, and when these two proteins interact, the luciferase fragments can reversibly associate and reconstitute enzyme activity. Though this technology has been used extensively in live eukaryotic cells, split luciferase complementation has not yet been applied to studies of dynamic protein-protein interactions in live bacteria. As proof of concept and to develop a new tool for studies of bacterial chemotaxis, fragments of Renilla luciferase (Rluc) were fused to the chemotaxis-associated response regulator CheY3 and its phosphatase CheZ in the enteric pathogen Vibrio cholerae. Luciferase activity was dependent on the presence of both CheY3 and CheZ fusion proteins, demonstrating the specificity of the assay. Furthermore, enzyme activity was markedly reduced in V. cholerae chemotaxis mutants, suggesting that this approach can measure defects in chemotactic signaling. However, attempts to measure changes in dynamic CheY3-CheZ interactions in response to various chemoeffectors were undermined by nonspecific inhibition of the full-length luciferase. These observations reveal an unexpected limitation of split Rluc complementation that may have implications for existing data and highlight the need for great caution when evaluating small molecule effects on dynamic protein-protein interactions using the split luciferase technology. PMID:22905225

Studies of dynamic protein-protein interactions in bacteria using Renilla luciferase complementation are undermined by nonspecific enzyme inhibition.

PubMed

Hatzios, Stavroula K; Ringgaard, Simon; Davis, Brigid M; Waldor, Matthew K

2012-01-01

The luciferase protein fragment complementation assay is a powerful tool for studying protein-protein interactions. Two inactive fragments of luciferase are genetically fused to interacting proteins, and when these two proteins interact, the luciferase fragments can reversibly associate and reconstitute enzyme activity. Though this technology has been used extensively in live eukaryotic cells, split luciferase complementation has not yet been applied to studies of dynamic protein-protein interactions in live bacteria. As proof of concept and to develop a new tool for studies of bacterial chemotaxis, fragments of Renilla luciferase (Rluc) were fused to the chemotaxis-associated response regulator CheY3 and its phosphatase CheZ in the enteric pathogen Vibrio cholerae. Luciferase activity was dependent on the presence of both CheY3 and CheZ fusion proteins, demonstrating the specificity of the assay. Furthermore, enzyme activity was markedly reduced in V. cholerae chemotaxis mutants, suggesting that this approach can measure defects in chemotactic signaling. However, attempts to measure changes in dynamic CheY3-CheZ interactions in response to various chemoeffectors were undermined by nonspecific inhibition of the full-length luciferase. These observations reveal an unexpected limitation of split Rluc complementation that may have implications for existing data and highlight the need for great caution when evaluating small molecule effects on dynamic protein-protein interactions using the split luciferase technology.

Site-targeted mutagenesis for stabilization of recombinant monoclonal antibody expressed in tobacco (Nicotiana tabacum) plants

PubMed Central

Hehle, Verena K.; Paul, Matthew J.; Roberts, Victoria A.; van Dolleweerd, Craig J.; Ma, Julian K.-C.

2016-01-01

This study examined the degradation pattern of a murine IgG1κ monoclonal antibody expressed in and extracted from transformed Nicotiana tabacum. Gel electrophoresis of leaf extracts revealed a consistent pattern of recombinant immunoglobulin bands, including intact and full-length antibody, as well as smaller antibody fragments. N-terminal sequencing revealed these smaller fragments to be proteolytic cleavage products and identified a limited number of protease-sensitive sites in the antibody light and heavy chain sequences. No strictly conserved target sequence was evident, although the peptide bonds that were susceptible to proteolysis were predominantly and consistently located within or near to the interdomain or solvent-exposed regions in the antibody structure. Amino acids surrounding identified cleavage sites were mutated in an attempt to increase resistance. Different Guy’s 13 antibody heavy and light chain mutant combinations were expressed transiently in N. tabacum and demonstrated intensity shifts in the fragmentation pattern, resulting in alterations to the full-length antibody-to-fragment ratio. The work strengthens the understanding of proteolytic cleavage of antibodies expressed in plants and presents a novel approach to stabilize full-length antibody by site-directed mutagenesis.—Hehle, V. K., Paul, M. J., Roberts, V. A., van Dolleweerd, C. J., Ma, J. K.-C. Site-targeted mutagenesis for stabilization of recombinant monoclonal antibody expressed in tobacco (Nicotiana tabacum) plants. PMID:26712217

DNA degradation and genetic analysis of empty puparia: genetic identification limits in forensic entomology.

PubMed

Mazzanti, Morena; Alessandrini, Federica; Tagliabracci, Adriano; Wells, Jeffrey D; Campobasso, Carlo P

2010-02-25

Puparial cases are common remnants of necrophagous flies in crime investigations. They usually represent the longest developmental time and, therefore, they can be very useful for the estimation of the post-mortem interval (PMI). However, before any PMI estimate, it is crucial to identify the species of fly eclosed from each puparium associated with the corpse. Morphological characteristics of the puparium are often distinctive enough to permit a species identification. But, even an accurate morphological analysis of empty puparia cannot discriminate among different species of closely related flies. Furthermore, morphological identification may be impossible if the fly puparia are poorly preserved or in fragments. This study explores the applicability of biomolecular techniques on empty puparia and their fragments for identification purposes. A total of 63 empty puparia of necrophagous Diptera resulting from forensic casework were examined. Samples were divided into three groups according to size, type and time of eclosion in order to verify whether the physical characteristics and puparia weathering can influence the amount of DNA extraction. The results suggest that a reliable genetic identification of forensically important flies may also be performed from empty puparia and/or their fragments. However, DNA degradation can deeply compromise the genetic analysis since the older the fly puparia, the smaller are the amplified fragments. 2009 Elsevier Ireland Ltd. All rights reserved.

A framework to predict the impacts of shale gas infrastructures on the forest fragmentation of an agroforest region.

PubMed

Racicot, Alexandre; Babin-Roussel, Véronique; Dauphinais, Jean-François; Joly, Jean-Sébastien; Noël, Pascal; Lavoie, Claude

2014-05-01

We propose a framework to facilitate the evaluation of the impacts of shale gas infrastructures (well pads, roads, and pipelines) on land cover features, especially with regards to forest fragmentation. We used a geographic information system and realistic development scenarios largely inspired by the PA (United States) experience, but adapted to a region of QC (Canada) with an already fragmented forest cover and a high gas potential. The scenario with the greatest impact results from development limited by regulatory constraints only, with no access to private roads for connecting well pads to the public road network. The scenario with the lowest impact additionally integrates ecological constraints (deer yards, maple woodlots, and wetlands). Overall the differences between these two scenarios are relatively minor, with <1 % of the forest cover lost in each case. However, large areas of core forests would be lost in both scenarios and the number of forest patches would increase by 13-21 % due to fragmentation. The pipeline network would have a much greater footprint on the land cover than access roads. Using data acquired since the beginning of the shale gas industry, we show that it is possible, within a reasonable time frame, to produce a robust assessment of the impacts of shale gas extraction. The framework we propose could easily be applied to other contexts or jurisdictions.

Fragment Based Strategies for Discovery of Novel HIV-1 Reverse Transcriptase and Integrase Inhibitors.

PubMed

Latham, Catherine F; La, Jennifer; Tinetti, Ricky N; Chalmers, David K; Tachedjian, Gilda

2016-01-01

Human immunodeficiency virus (HIV) remains a global health problem. While combined antiretroviral therapy has been successful in controlling the virus in patients, HIV can develop resistance to drugs used for treatment, rendering available drugs less effective and limiting treatment options. Initiatives to find novel drugs for HIV treatment are ongoing, although traditional drug design approaches often focus on known binding sites for inhibition of established drug targets like reverse transcriptase and integrase. These approaches tend towards generating more inhibitors in the same drug classes already used in the clinic. Lack of diversity in antiretroviral drug classes can result in limited treatment options, as cross-resistance can emerge to a whole drug class in patients treated with only one drug from that class. A fresh approach in the search for new HIV-1 drugs is fragment-based drug discovery (FBDD), a validated strategy for drug discovery based on using smaller libraries of low molecular weight molecules (<300 Da) screened using primarily biophysical assays. FBDD is aimed at not only finding novel drug scaffolds, but also probing the target protein to find new, often allosteric, inhibitory binding sites. Several fragment-based strategies have been successful in identifying novel inhibitory sites or scaffolds for two proven drug targets for HIV-1, reverse transcriptase and integrase. While any FBDD-generated HIV-1 drugs have yet to enter the clinic, recent FBDD initiatives against these two well-characterised HIV-1 targets have reinvigorated antiretroviral drug discovery and the search for novel classes of HIV-1 drugs.

How Networks of Informal Trails Cause Landscape Level Damage to Vegetation.

PubMed

Barros, Agustina; Marina Pickering, Catherine

2017-07-01

When visitors are not constrained to remain on formal trails, informal trail networks can develop and damage plant communities in protected areas. These networks can form in areas with low growing vegetation, where formal trails are limited, where there is limited regulation and where vegetation is slow to recover once disturbed. To demonstrate the extent of impacts from unregulated recreational use, we assessed damage to alpine vegetation by hikers and pack animals in the highest protected area in the southern Hemisphere: Aconcagua Park, in the Andes. Within the 237 ha area surveyed in the Horcones Valley, over 19 km of trails were found, nearly all of which (94%) were informal. This network of trails resulted in the direct loss of 11.5 ha of vegetation and extensive fragmentation of alpine meadows (21 fragments) and steppe vegetation (68 fragments). When levels of disturbance off these trails were quantified using rapid visual assessments, 81% of 102 randomly located plots showed evidence of disturbance, with the severity of disturbance greatest close to trails. As a result, vegetation in 90% of the Valley has been damaged by visitor use, nearly all of it from unregulated use. These results highlight the extent to which informal trails and trampling off-trail can cause landscape damage to areas of high conservation value, and hence the importance of better regulation of visitor use. The methodology used for off-trail impact assessment can be easily applied or adapted for other popular protected areas where trampling off-trail is also an issue.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cetrangolo, Giovanni Paolo, E-mail: giovanni.cetrangolo@unimol.it; Arcaro, Alessia, E-mail: alessia.arcaro@unimol.it; Lepore, Alessio, E-mail: alessiolepore@alice.it

Highlights: • A carboxy-terminal fragment (residues 2515–2750) was isolated from a low-iodine bTg. • Post-translational status of 8 tyrosines in bTg region 2515–2750 was assessed by MS. • Tyr2522 of bovine Tg is an interspecifically conserved hormonogenic donor site. • Propensities of Tyr residues to mono or diiodination optimize T3 yield from Tyr2748. - Abstract: A tryptic fragment (b5{sub TR,NR}), encompassing residues 2515–2750, was isolated from a low-iodine (0.26% by mass) bovine thyroglobulin, by limited proteolysis with trypsin and preparative, continuous-elution SDS–PAGE. The fragment was digested with Asp-N endoproteinase and analyzed by reverse-phase HPLC electrospray ionization quadrupole time-of-flight mass spectrometry,more » revealing the formation of: 3-monoiodotyrosine and dehydroalanine from Tyr2522; 3-monoiodotyrosine from Tyr2555 and Tyr2569; 3-monoiodotyrosine and 3,5-diiodotyrosine from Tyr2748. The data presented document, by direct mass spectrometric identifications, efficient iodophenoxyl ring transfer from monoiodinated hormonogenic donor Tyr2522 and efficient mono- and diiodination of hormonogenic acceptor Tyr2748, under conditions which permitted only limited iodination of Tyr2555 and Tyr2569, in low-iodine bovine thyroglobulin. The present study thereby provides: (1) a rationale for the preferential synthesis of T3 at the carboxy-terminal end of thyroglobulin, at low iodination level; (2) confirmation for the presence of an interspecifically conserved hormonogenic donor site in the carboxy-terminal domain of thyroglobulin; (3) solution for a previous uncertainty, concerning the precise location of such donor site in bovine thyroglobulin.« less

Development of a high yielding E. coli periplasmic expression system for the production of humanized Fab' fragments.

PubMed

Ellis, Mark; Patel, Pareshkumar; Edon, Marjory; Ramage, Walter; Dickinson, Robert; Humphreys, David P

2017-01-01

Humanized Fab' fragments may be produced in the periplasm of Escherichia coli but can be subject to degradation by host cell proteases. In order to increase Fab' yield and reduce proteolysis we developed periplasmic protease deficient strains of E. coli. These strains lacked the protease activity of Tsp, protease III and DegP. High cell density fermentations indicated Tsp deficient strains increased productivity two fold but this increase was accompanied by premature cell lysis soon after the induction of Fab' expression. To overcome the reduction in cell viability we introduced suppressor mutations into the spr gene. The mutations partially restored the wild type phenotype of the cells. Furthermore, we coexpressed a range of periplasmic chaperone proteins with the Fab', DsbC had the most significant impact, increasing humanized Fab' production during high cell density fermentation. When DsbC coexpression was combined with a Tsp deficient spr strain we observed an increase in yield and essentially restored "wild type" cell viability. We achieved a final periplasmic yield of over 2.4g/L (final cell density OD 600 105), 40 h post Fab' induction with minimal cell lysis.The data suggests that proteolysis, periplasm integrity, protein folding and disulphide bond formation are all potential limiting steps in the production of Fab' fragments in the periplasm of E. coli. In this body of work, we have addressed these limiting steps by utilizing stabilized protease deficient strains and chaperone coexpression. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:212-220, 2017. © 2016 American Institute of Chemical Engineers.

Competition for dead trees between humans and aye-ayes (Daubentonia madagascariensis) in central eastern Madagascar.

PubMed

Miller, Rose T; Raharison, Jean-Luc; Irwin, Mitchell T

2017-04-01

The destruction and degradation of forest habitats are major threats to the sustainability of lemur populations in Madagascar. Madagascan landscapes often contain forest fragments that represent refuges for native fauna, while also being used for firewood and timber by local human populations. As undisturbed forest becomes increasingly scarce, understanding resource competition between humans and wildlife in disturbed habitats will be increasingly important. We tested the hypothesis that Malagasy and aye-ayes (Daubentonia madagascariensis) compete for the limited number of dead trees in rainforest fragments at Tsinjoarivo, Madagascar. We surveyed 2.16 ha within five fragments (range 5-228 ha) surrounding human settlements to quantify the density of dead trees and traces of both human and aye-aye activity. Neither aye-aye nor human traces were distributed according to the availability of particular trees species, and aye-ayes and Malagasy apparently preferred several different species. Although overlap was recorded in tree species used, human use tended to be positively correlated with a species' desirability as firewood, while a negative relationship was seen for aye-ayes. Both consumers used trees of similar diameter at breast height, but those used by aye-ayes tended to be older, suggesting that human use might precede usefulness for aye-ayes. Finally, the density of dead trees and aye-aye traces were highest in smaller fragments, but human traces did not vary across fragment size. Although further study is needed to better quantify the aye-aye diet in this region, these data suggest that aye-ayes and local people compete for dead trees, and this competition could constitute a pressure on aye-aye populations.

Subterranean fragmentation of magma during conduit initiation and evolution in the shallow plumbing system of the small-volume Jagged Rocks volcanoes (Hopi Buttes Volcanic Field, Arizona, USA)

NASA Astrophysics Data System (ADS)

Re, Giuseppe; White, James D. L.; Muirhead, James D.; Ort, Michael H.

2016-08-01

Monogenetic volcanoes have limited magma supply and lack long-lived sustained magma plumbing systems. They erupt once, often from multiple vents and sometimes over several years, and are rarely or never re-activated. Eruptive behavior is very sensitive to physical processes (e.g., volatile exsolution, magma-water interaction) occurring in the later stages of magma ascent at shallow crustal depths (<1 km), which yield a spectrum of eruptive styles including weak to moderate explosive activity, violent phreatomagmatism, and lava effusion. Jagged Rocks Complex in the late Miocene Hopi Buttes Volcanic field (Arizona, USA) exposes the frozen remnants of the feeding systems for one or a few monogenetic volcanoes. It provides information on how a shallow magmatic plumbing system evolved within a stable non-marine sedimentary basin, and the processes by which magma flowing through dikes fragmented and conduits were formed. We have identified three main types of fragmental deposits, (1) buds (which emerge from dikes), (2) pyroclastic massifs, and (3) diatremes; these represent three different styles and intensities of shallow-depth magma fragmentation. They may develop successively and at different sites during the evolution of a monogenetic volcano. The deposits consist of a mixture of pyroclasts with varying degrees of welding and country-rock debris in various proportions. Pyroclasts are commonly welded together, but also reveal in places features consistent with phreatomagmatism, such as blocky shapes, dense groundmasses, and composite clasts (loaded and cored). The extent of fragmentation and the formation of subterranean open space controlled the nature of the particles and the architecture and geometry of these conduit structures and their deposits.

First Observation of ^60Ge and ^64Se

NASA Astrophysics Data System (ADS)

Stolz, Andreas

2004-10-01

The neutron-deficient nuclei ^60Ge and ^64Se were observed for the first time following the fragmentation of ^78Kr. This is the first observation of new proton-rich nuclei below Z=50 in over three years. A primary beam of 140-MeV/nucleon ^78Kr was produced using the Coupled Cyclotron Facility and fragmented in a Be target. The secondary neutron-deficient fragments were separated by the A1900 fragment separator [1] and stopped in a stack of silicon PIN diodes. The fragments were identified by a measurement of energy loss, total energy, and time-of-flight. ^60Ge is the last nucleus along the proton dripline lighter than cadmium which is predicted to be bound (S_2p = 50 ± 240 keV) by the latest atomic mass evaluation [2] and which has not yet been observed. Theoretical calculations predict ^60Ge to be bound with respect to two-proton emission by 167(141) keV [3] and 630 keV [4], while ^64Se is predicted not to be bound [2]. >From the number of observed events of ^60Ge and ^64Se the production cross section and lifetime will be estimated. No events of ^59Ga and ^63As were observed confirming that these nuclei are unbound with respect to proton emission and upper limits of the lifetime can be established. [1] D. J. Morrissey, B. M. Sherrill, M. Steiner, A. Stolz, and I. Wiedenhover, Nucl. Instrum. Methods Phys. Res. B 204, 90 (2003). [2] G. Audi, A. H. Wapstra, and C. Thibault, Nucl. Phys. A729, 337 (2003). [3] W. E. Ormand, Phys. Rev. C 55, 2407 (1997). [4] M.V. Stoitsov, J. Dobaczewski, W. Nazarewicz, S. Pittel, and D. J. Dean, Phys. Rev. C 68, 054312 (2003).

Electron ionization of SiCl4

NASA Astrophysics Data System (ADS)

King, Simon J.; Price, Stephen D.

2011-02-01

Relative partial ionization cross sections (PICS) for the formation of fragment ions following electron ionization of SiCl4, in the electron energy range 30-200 eV, have been determined using time-of-flight mass spectrometry coupled with an ion coincidence technique. By this method, the contributions to the yield of each fragment ion from dissociative single, double, and triple ionization, are distinguished. These yields are quantified in the form of relative precursor-specific PICS, which are reported here for the first time for SiCl4. For the formation of singly charged ionic fragments, the low-energy maxima appearing in the PICS curves are due to contributions from single ionization involving predominantly indirect ionization processes, while contributions to the yields of these ions at higher electron energies are often dominated by dissociative double ionization. Our data, in the reduced form of relative PICS, are shown to be in good agreement with a previous determination of the PICS of SiCl4. Only for the formation of doubly charged fragment ions are the current relative PICS values lower than those measured in a previous study, although both datasets agree within combined error limits. The relative PICS data presented here include the first quantitative measurements of the formation of Cl2+ fragment ions and of the formation of ion pairs via dissociative double ionization. The peaks appearing in the 2D ion coincidence data are analyzed to provide further information concerning the mechanism and energetics of the charge-separating dissociations of SiCl42+. The lowest energy dicationic precursor state, leading to SiCl3+ + Cl+ formation, lies 27.4 ± 0.3 eV above the ground state of SiCl4 and is in close agreement with a calculated value of the adiabatic double ionization energy (27.3 eV).

Electron ionization of SiCl4.

PubMed

King, Simon J; Price, Stephen D

2011-02-21

Relative partial ionization cross sections (PICS) for the formation of fragment ions following electron ionization of SiCl(4), in the electron energy range 30-200 eV, have been determined using time-of-flight mass spectrometry coupled with an ion coincidence technique. By this method, the contributions to the yield of each fragment ion from dissociative single, double, and triple ionization, are distinguished. These yields are quantified in the form of relative precursor-specific PICS, which are reported here for the first time for SiCl(4). For the formation of singly charged ionic fragments, the low-energy maxima appearing in the PICS curves are due to contributions from single ionization involving predominantly indirect ionization processes, while contributions to the yields of these ions at higher electron energies are often dominated by dissociative double ionization. Our data, in the reduced form of relative PICS, are shown to be in good agreement with a previous determination of the PICS of SiCl(4). Only for the formation of doubly charged fragment ions are the current relative PICS values lower than those measured in a previous study, although both datasets agree within combined error limits. The relative PICS data presented here include the first quantitative measurements of the formation of Cl(2) (+) fragment ions and of the formation of ion pairs via dissociative double ionization. The peaks appearing in the 2D ion coincidence data are analyzed to provide further information concerning the mechanism and energetics of the charge-separating dissociations of SiCl(4) (2+). The lowest energy dicationic precursor state, leading to SiCl(3) (+) + Cl(+) formation, lies 27.4 ± 0.3 eV above the ground state of SiCl(4) and is in close agreement with a calculated value of the adiabatic double ionization energy (27.3 eV).

A century of landscape disturbance and urbanization of the San Francisco Bay region affects the present-day genetic diversity of the California Ridgway’s rail (Rallus obsoletus obsoletus)

USGS Publications Warehouse

Wood, Dustin A.; Bui, Thuy-Vy D.; Overton, Cory T.; Vandergast, Amy; Casazza, Michael L.; Hull, Joshua M.; Takekawa, John Y.

2016-01-01

Fragmentation and loss of natural habitat have important consequences for wild populations and can negatively affect long-term viability and resilience to environmental change. Salt marsh obligate species, such as those that occupy the San Francisco Bay Estuary in western North America, occupy already impaired habitats as result of human development and modifications and are highly susceptible to increased habitat loss and fragmentation due to global climate change. We examined the genetic variation of the California Ridgway’s rail (Rallus obsoletus obsoletus), a state and federally endangered species that occurs within the fragmented salt marsh of the San Francisco Bay Estuary. We genotyped 107 rails across 11 microsatellite loci and a single mitochondrial gene to estimate genetic diversity and population structure among seven salt marsh fragments and assessed demographic connectivity by inferring patterns of gene flow and migration rates. We found pronounced genetic structuring among four geographically separate genetic clusters across the San Francisco Bay. Gene flow analyses supported a stepping stone model of gene flow from south-to-north. However, contemporary gene flow among the regional embayments was low. Genetic diversity among occupied salt marshes and genetic clusters were not significantly different. We detected low effective population sizes and significantly high relatedness among individuals within salt marshes. Preserving genetic diversity and connectivity throughout the San Francisco Bay may require attention to salt marsh restoration in the Central Bay where habitat is both most limited and most fragmented. Incorporating periodic genetic sampling into the management regime may help evaluate population trends and guide long-term management priorities.

Identification of substances migrating from plastic baby bottles using a combination of low-resolution and high-resolution mass spectrometric analysers coupled to gas and liquid chromatography.

PubMed

Onghena, Matthias; Van Hoeck, Els; Van Loco, Joris; Ibáñez, María; Cherta, Laura; Portolés, Tania; Pitarch, Elena; Hernandéz, Félix; Lemière, Filip; Covaci, Adrian

2015-11-01

This work presents a strategy for elucidation of unknown migrants from plastic food contact materials (baby bottles) using a combination of analytical techniques in an untargeted approach. First, gas chromatography (GC) coupled to mass spectrometry (MS) in electron ionisation mode was used to identify migrants through spectral library matching. When no acceptable match was obtained, a second analysis by GC-(electron ionisation) high resolution mass spectrometry time of flight (TOF) was applied to obtain accurate mass fragmentation spectra and isotopic patterns. Databases were then searched to find a possible elemental composition for the unknown compounds. Finally, a GC hybrid quadrupole-TOF-MS with an atmospheric pressure chemical ionisation source was used to obtain the molecular ion or the protonated molecule. Accurate mass data also provided additional information on the fragmentation behaviour as two acquisition functions with different collision energies were available (MS(E) approach). In the low-energy function, limited fragmentation took place, whereas for the high-energy function, fragmentation was enhanced. For less volatile unknowns, ultra-high pressure liquid chromatography-quadrupole-TOF-MS was additionally applied. Using a home-made database containing common migrating compounds and plastic additives, tentative identification was made for several positive findings based on accurate mass of the (de)protonated molecule, product ion fragments and characteristic isotopic ions. Six illustrative examples are shown to demonstrate the modus operandi and the difficulties encountered during identification. The combination of these techniques was proven to be a powerful tool for the elucidation of unknown migrating compounds from plastic baby bottles. Copyright © 2015 John Wiley & Sons, Ltd.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sim, Eunji; Kim, Min-Cheol; Burke, Kieron

We investigate dissociation of diatomic molecules using standard density functional theory (DFT) and density-corrected density functional theory (DC-DFT) compared with CCSD(T) results as reference. The results show the difference between the HOMO values of dissociated atomic species often can be used as an indicator whether DFT would predict the correct dissociation limit. DFT predicts incorrect dissociation limits and charge distribution in molecules or molecular ions when the fragments have large HOMO differences, while DC-DFT and CCSD(T) do not. The criteria for large HOMO difference is about 2 ∼ 4 eV.

Challenging the Limits of Critique in Education through Morin's Paradigm of Complexity

ERIC Educational Resources Information Center

Alhadeff-Jones, Michel

2010-01-01

The position adopted in this paper is inspired by Edgar Morin's paradigm of complexity and his critique of scientific and philosophical forms of reductionism. This paper is based on research focusing on the diversity of conceptions of critique developed in academic discourses. It aims to challenge the fragmentation and the reduction framing the…

Barriers, invasion, and conservation of native salmonids in coldwater streams [Box 18.2

Treesearch

Bruce Rieman; Michael Young; Kurt Fausch; Jason Dunham; Douglas Peterson

2010-01-01

Habitat loss and fragmentation are threats to persistence of many native fish populations. Invading nonnative species that may restrict or displace native species are also important. These two issues are particularly relevant for native salmonids that are often limited to remnant habitats in cold, headwater streams. On the surface, reversing threats to native fishes...

Cytogeography of Larrea tridentata at the Chihuahuan-Sonoran Desert ecotone

Treesearch

Robert G. Laport; Robert L. Minckley

2013-01-01

The long separation of the Chihuahuan and Sonoran Deserts is reflected in the high species richness and endemism of their floras. Although many endemic species from both deserts reach their distributional limits where the Sierra Madre Occidental massif fragments into smaller mountain complexes in northern Mexico and adjoining areas of the United States, indicator...

Telomere erosion varies during in vitro aging of normal human fibroblasts from young and adult donors.

PubMed

Figueroa, R; Lindenmaier, H; Hergenhahn, M; Nielsen, K V; Boukamp, P

2000-06-01

The life span of normal fibroblasts in vitro (Hayflick limit) depends on donor age, and telomere shortening has been proposed as a potential mechanism. By quantitative fluorescence in situ hybridization and Southern blot analysis, we show progressive telomere loss to about 5 kb mean telomere restriction fragment length in fibroblasts from two adult donors within 40 population doublings, whereas in fibroblasts from two infant donors, telomere erosion is reduced, leaving a mean telomere restriction fragment length of approximately 7 kb at senescence (after approximately 60 population doublings). Aging of fibroblasts from both infant and adult donors was not accompanied by chromosomal abnormalities but was correlated with increased telomere repeat-binding factor 2 expression at both the protein and transcriptional level.

Open Reduction in Pediatric Condylar Fracture.

PubMed

Barbosa, Alexandre Agostini; Mariano, Ronaldo Célio

2017-05-01

Facial fractures in children are rare. Lack of pneumatization, fat pockets, mixed dentition, contribute to the elasticity and bone stability. When mandibular fractures occur in children, most often involve the condyle by indirect trauma. Such fractures are the center of discussion on the form of treatment if this should be performed conservatively, or held the reduction and fixation of the fracture with surgical exposure of the fragments. In condylar fractures in children, in most patients, the proposed treatment is closed reduction. Treatment with open reduction and fixation has its specific indications. In this case, the authors report a patient cycling accident victim, with cut-contusion injury in ment with limited mouth opening and left condylar fracture with medial rotation. The treatment was the reduction and fixation of fragments by open process.

Electronic energy transfer: Localized operator partitioning of electronic energy in composite quantum systems

NASA Astrophysics Data System (ADS)

Khan, Yaser; Brumer, Paul

2012-11-01

A Hamiltonian based approach using spatially localized projection operators is introduced to give precise meaning to the chemically intuitive idea of the electronic energy on a quantum subsystem. This definition facilitates the study of electronic energy transfer in arbitrarily coupled quantum systems. In particular, the decomposition scheme can be applied to molecular components that are strongly interacting (with significant orbital overlap) as well as to isolated fragments. The result defines a consistent electronic energy at all internuclear distances, including the case of separated fragments, and reduces to the well-known Förster and Dexter results in their respective limits. Numerical calculations of coherent energy and charge transfer dynamics in simple model systems are presented and the effect of collisionally induced decoherence is examined.

Selection of stable scFv antibodies by phage display.

PubMed

Brockmann, Eeva-Christine

2012-01-01

ScFv fragments are popular recombinant antibody formats but often suffer from limited stability. Phage display is a powerful tool in antibody engineering and applicable also for stability selection. ScFv variants with improved stability can be selected from large randomly mutated phage displayed libraries with a specific antigen after the unstable variants have been inactivated by heat or GdmCl. Irreversible scFv denaturation, which is a prerequisite for efficient selection, is achieved by combining denaturation with reduction of the intradomain disulfide bonds. Repeated selection cycles of increasing stringency result in enrichment of stabilized scFv fragments. Procedures for constructing a randomly mutated scFv library by error-prone PCR and phage display selection for enrichment of stable scFv antibodies from the library are described here.

RETENTION OF POLYURETHANE FOAM FRAGMENTS DURING VAC-THERAPY, A COMPLICATION TO BE CONSIDERED

PubMed Central

Dessy, L.A.; Serratore, F.; Corrias, F.; Parisi, P.; Mazzocchi, M.; Carlesimo, B.

2013-01-01

VAC-therapy is a closed-loop, non-invasive active system, characterised by a controlled and localized negative pressure applied on porous polyurethane absorbent foams. It promotes healing of acute and chronic wounds. Therapeutic effects of VAC-therapy have been proved and demonstrated, however this method can have some disadvantages. Even if it is a quite versatile device, only qualified medical/paramedical personnel should use it in order to avoid possible complications that can occur after an improper application. In this report, 11 cases of foam fragment retention within the wound are presented. This rare complication did not promote healing, but further hindered it. On the basis of our experience, it is mandatory to define indications, benefits and limitations of the VAC-therapy. PMID:23590296

Mass-spectrometric identification of primary biological particle markers and application to pristine submicron aerosol measurements in Amazonia

NASA Astrophysics Data System (ADS)

Schneider, J.; Freutel, F.; Zorn, S. R.; Chen, Q.; Farmer, D. K.; Jimenez, J. L.; Martin, S. T.; Artaxo, P.; Wiedensohler, A.; Borrmann, S.

2011-11-01

The detection of primary biological material in submicron aerosol by means of thermal desorption/electron impact ionization aerosol mass spectrometry was investigated. Mass spectra of amino acids, carbohydrates, small peptides, and proteins, all of which are key building blocks of biological particles, were recorded in laboratory experiments. Several characteristic marker fragments were identified. The intensity of the marker signals relative to the total organic mass spectrum allows for an estimation of the content of primary biological material in ambient organic aerosol. The developed method was applied to mass spectra recorded during AMAZE-08, a field campaign conducted in the pristine rainforest of the central Amazon Basin, Brazil, during the wet season of February and March 2008. The low abundance of identified marker fragments places upper limits of 7.5% for amino acids and 5.6% for carbohydrates on the contribution of primary biological aerosol particles (PBAP) to the submicron organic aerosol mass concentration during this time period. Upper limits for the absolute submicron concentrations for both compound classes range from 0.01 to 0.1 μg m-3. Carbohydrates and proteins (composed of amino acids) make up for about two thirds of the dry mass of a biological cell. Thus, our findings suggest an upper limit for the PBAP mass fraction of about 20% to the submicron organic aerosol measured in Amazonia during AMAZE-08.

Crossed and Locked Quotes in a Multi-Market Simulation

PubMed Central

Todd, Andrew; Beling, Peter; Scherer, William

2016-01-01

Financial markets are often fragmented, introducing the possibility that quotes in identical securities may become crossed or locked. There are a number of theoretical explanations for the existence of crossed and locked quotes, including competition, simultaneous actions, inattentiveness, fee structure and market access. In this paper, we perform a simulation experiment designed to examine the effect of simple order routing procedures on the properties of a fragmented market consisting of a single security trading in two independent limit order books. The quotes in the two markets are connected solely by the routing decision of the market participants. We report on the health of the consolidated market as measured by the duration of crossed and locked states, as well as the spread and the volatility of transaction prices in the consolidated market. We aim to quantify exactly how the prevalence of order routing among a population of market participants affects properties of the consolidated market. Our model contributes to the zero-intelligence literature by treating order routing as an experimental variable. Additionally, we introduce a parsimonious heuristic for limit order routing, allowing us to study the effects of both market order routing and limit order routing. Our model refines intuition for the sometimes subtle relationships between the prevalence of order routing and various market measures. Our model also provides a benchmark for more complex agent-based models. PMID:26959416

Crossed and Locked Quotes in a Multi-Market Simulation.

PubMed

Todd, Andrew; Beling, Peter; Scherer, William

2016-01-01

Financial markets are often fragmented, introducing the possibility that quotes in identical securities may become crossed or locked. There are a number of theoretical explanations for the existence of crossed and locked quotes, including competition, simultaneous actions, inattentiveness, fee structure and market access. In this paper, we perform a simulation experiment designed to examine the effect of simple order routing procedures on the properties of a fragmented market consisting of a single security trading in two independent limit order books. The quotes in the two markets are connected solely by the routing decision of the market participants. We report on the health of the consolidated market as measured by the duration of crossed and locked states, as well as the spread and the volatility of transaction prices in the consolidated market. We aim to quantify exactly how the prevalence of order routing among a population of market participants affects properties of the consolidated market. Our model contributes to the zero-intelligence literature by treating order routing as an experimental variable. Additionally, we introduce a parsimonious heuristic for limit order routing, allowing us to study the effects of both market order routing and limit order routing. Our model refines intuition for the sometimes subtle relationships between the prevalence of order routing and various market measures. Our model also provides a benchmark for more complex agent-based models.

Secondary Mineral Deposits and Evidence of Past Seismicity and Heating of the Proposed Repository Horizon at Yucca Mountain, Nevada

USGS Publications Warehouse

Whelan, Josheph F.

2004-01-01

The Drift Degradation Analysis (DDA) (BSC, 2003) for the proposed high-level radioactive waste repository at Yucca Mountain, Nevada, describes model simulations of the effects of pre- and post-closure seismicity and waste-induced heating on emplacement drifts. Based on probabilistic seismic hazard analyses of the intensity and frequency of future seismic events in the region (CRWMS M&O, 1998), the DDA concludes that future seismicity will lead to substantial damage to emplacement drifts, particularly those in the lithophysal tuffs, where some simulations predict complete collapse of the drift walls. Secondary mineral studies conducted by the U.S. Geological Survey since 1995 indicate that secondary calcite and silica have been deposited in some fractures and lithophysal cavities in the unsaturated zone (UZ) at Yucca Mountain during at least the past 10 million years (m.y.), and probably since the tuffs cooled to less than 100?C. Tuff fragments, likely generated by past seismic activity, have commonly been incorporated into the secondary mineral depositional sequences. Preliminary observations indicate that seismic activity has generated few, if any, tuff fragments during the last 2 to 4 m.y., which may be inconsistent with the predictions of drift-wall collapse described in the DDA. Whether or not seismicity-induced tuff fragmentation occurring at centimeter to decimeter scales in the fracture and cavity openings relates directly to failure of tuff walls in the 5.5-m-diameter waste emplacement drifts, the deposits do provide a potential record of the spatial and temporal distribution of tuff fragments in the UZ. In addition, the preservation of weakly attached coatings and (or) delicate, upright blades of calcite in the secondary mineral deposits provides an upper limit for ground motion during the late stage of deposition that might be used as input to future DDA simulations. Finally, bleaching and alteration at a few of the secondary mineral sites indicate that they were subjected to heated gases at approximately the temperatures expected from waste emplacement. These deposits provide at least limited textural and mineralogic analogs for waste-induced, high-humidity thermal alteration of emplacement drift wall rocks.

Comparison of CHROMagar, polymerase chain reaction-restriction fragment length polymorphism, and polymerase chain reaction-fragment size for the identification of Candida species.

PubMed

Jafari, Zahra; Motamedi, Marjan; Jalalizand, Nilufar; Shokoohi, Gholam R; Charsizadeh, Arezu; Mirhendi, Hossein

2017-09-01

The epidemiological alteration in the distribution of Candida species, as well as the significantly increasing trend of either intrinsic or acquired resistance of some of these fungi highlights the need for a reliable method for the identification of the species. Polymerase chain reaction (PCR) is one of the methods facilitating the quick and precise identification of Candida species. The aim of this study was to compare the efficiency of CHROMagar, PCR-restriction fragment length polymorphism (PCR-RFLP), and PCR-fragment size polymorphism (PCR-FSP) assays in the identification of Candida species to determine the benefits and limitations of these methods. This study was conducted on 107 Candida strains, including 20 standard strains and 87 clinical isolates. The identification of the isolates was accomplished by using CHROMagar as a conventional method. The PCR-RFLP assay was performed on the entire internal transcribed spacer (ITS) region of ribosomal DNA (rDNA), and the consequent enzymatic digestion was compared with PCR-FSP results in which ITS1 and ITS2 regions were separately PCR amplified. In both molecular assays, yeast identification was carried out through the specific electrophoretic profiles of the PCR products. According to the results, the utilization of CHROMagar resulted in the identification of 29 (33.3%) Candida isolates, while the PCR-RFLP and PCR-FSP facilitated the identification of 83 (95.4%) and 80 (91.9%) clinical isolates, respectively. The obtained concordances between CHROMagar and PCR-RFLP, between CHROMagar and PCR-FSP, as well as between PCR-RFLP and PCR-FSP were 0.23, 0.20, and 0.77, respectively. The recognition of the benefits and limitations of PCR methods allows for the selection of the most efficient technique for a fast and correct differentiation. The PCR-RFLP and PCR-FSP assays had satisfactory concordance. The PCR-FSP provides a rapid, technically simple, and cost-effective method for the identification of Candida species. Nevertheless, to accurately differentiate among the taxonomically related species, PCR-RFLP should be implemented.

How well does the Friends-of-Friends algorithm recover group properties from galaxy catalogues limited in both distance and luminosity?

NASA Astrophysics Data System (ADS)

Duarte, Manuel; Mamon, Gary A.

2014-05-01

The specific star formation rates of galaxies are influenced both by their mass and by their environment. Moreover, the mass function of groups and clusters serves as a powerful cosmological tool. It is thus important to quantify the accuracy to which group properties are extracted from redshift surveys. We test here the Friends-of-Friends (FoF) grouping algorithm, which depends on two linking lengths (LLs), plane-of-sky and line-of-sight (LOS), normalized to the mean nearest neighbour separation of field galaxies. We argue, on theoretical grounds, that LLs should be b⊥ ≃ 0.11, and b∥ ≈ 1.3 to recover 95 per cent of all galaxies with projected radii within the virial radius r200 and 95 per cent of the galaxies along the LOS. We then predict that 80 to 90 per cent of the galaxies in FoF groups should lie within their parent real-space groups (RSGs), defined within their virial spheres. We test the FoF extraction for 16 × 16 pairs of LLs, using subsamples of galaxies, doubly complete in distance and luminosity, of a flux-limited mock Sloan Digital Sky Survey (SDSS) galaxy catalogue. We find that massive RSGs are more prone to fragmentation, while the fragments typically have low estimated mass, with typically 30 per cent of groups of low and intermediate estimated mass being fragments. Group merging rises drastically with estimated mass. For groups of three or more galaxies, galaxy completeness and reliability are both typically better than 80 per cent (after discarding the fragments). Estimated masses of extracted groups are biased low, by up to a factor 4 at low richness, while the inefficiency of mass estimation improves from 0.85 dex to 0.2 dex when moving from low to high multiplicity groups. The optimal LLs depend on the scientific goal for the group catalogue. We propose b⊥ ≃ 0.07, with b∥ ≃ 1.1 for studies of environmental effects, b∥ ≃ 2.5 for cosmographic studies and b∥ ≃ 5 for followups of individual groups.

Pollen-limited reproduction in blue oak: Implications for wind pollination in fragmented populations

USGS Publications Warehouse

Knapp, E.E.; Goedde, M.A.; Rice, K.J.

2001-01-01

Human activities are fragmenting forests and woodlands worldwide, but the impact of reduced tree population densities on pollen transfer in wind-pollinated trees is poorly understood. In a 4-year study, we evaluated relationships among stand density, pollen availability, and seed production in a thinned and fragmented population of blue oak (Quercus douglasii). Geographic coordinates were established and flowering interval determined for 100 contiguous trees. The number of neighboring trees within 60 m that released pollen during each tree's flowering period was calculated and relationships with acorn production explored using multiple regression. We evaluated the effects of female flower production, average temperature, and relative humidity during the pollination period, and number of pollen-producing neighbors on individual trees' acorn production. All factors except temperature were significant in at least one of the years of our study, but the combination of factors influencing acorn production varied among years. In 1996, a year of large acorn crop size, acorn production was significantly positively associated with number of neighboring pollen producers and density of female flowers. In 1997, 1998, and 1999, many trees produced few or no acorns, and significant associations between number of pollen-producing neighbors and acorn production were only apparent among moderately to highly reproductive trees. Acorn production by these reproductive trees in 1997 was significantly positively associated with number of neighboring pollen producers and significantly negatively associated with average relative humidity during the pollination period. In 1998, no analysis was possible, because too few trees produced a moderate to large acorn crop. Only density of female flowers was significantly associated with acorn production of moderately to highly reproductive trees in 1999. The effect of spatial scale was also investigated by conducting analyses with pollen producers counted in radii ranging from 30 m to 80 m. The association between number of pollen-producing neighbors and acorn production was strongest when neighborhood sizes of 60 m or larger were considered. Our results suggest that fragmentation and thinning of blue oak woodlands may reduce pollen availability and limit reproduction in this wind-pollinated species.

Demonstrating the effectiveness of body armour: a pilot prospective computerised surface wound mapping trial performed at the Role 3 hospital in Afghanistan.

PubMed

Breeze, Johno; Allanson-Bailey, L S; Hepper, A E; Midwinter, M J

2015-03-01

Modern body armour clearly reduces injury incidence and severity, but evidence to actually objectively demonstrate this effect is scarce. Although the Joint Theatre Trauma Registry (JTTR) alone cannot relate injury pattern to body armour coverage, the addition of computerised Surface Wound Mapping (SWM) may enable this utility. Surface wound locations of all UK and NATO coalition soldiers, Afghan National Army and Police and local nationals injured by explosively propelled fragments and treated in the Role 3 UK-led Field Hospital in Camp Bastion, Afghanistan, between 8 July and 20 October 2012 were prospectively recorded. The Abbreviated Injury Scores (AIS) and relative risk of casualties sustaining injuries under a type of body armour were compared with those that did not wear that armour. Casualties wearing a combat helmet were 2.7 times less likely to sustain a fragmentation wound to the head than those that were unprotected (mean AIS of 2.9 compared with 4.1). Casualties wearing a body armour vest were 4.1 times less likely to sustain a fragmentation wound to the chest or abdomen than those that were unprotected (mean AIS of 2.9 compared with 3.9). Casualties wearing pelvic protection were 10 times less likely to sustain a fragmentation wound to the pelvis compared with those that were unprotected (mean AIS of 3.4 compared with 3.9). Computerised SWM has objectively demonstrated the ability of body armour worn on current operations in Afghanistan to reduce wound incidence and severity. We recognise this technique is limited in that it only records the surface wound location and may be specific to this conflict. However, gathering electronic SWM at the same time as recording injuries for the JTTR was simple, required little extra time and therefore we would recommend its collection during future conflicts. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Validity and validation of expert (Q)SAR systems.

PubMed

Hulzebos, E; Sijm, D; Traas, T; Posthumus, R; Maslankiewicz, L

2005-08-01

At a recent workshop in Setubal (Portugal) principles were drafted to assess the suitability of (quantitative) structure-activity relationships ((Q)SARs) for assessing the hazards and risks of chemicals. In the present study we applied some of the Setubal principles to test the validity of three (Q)SAR expert systems and validate the results. These principles include a mechanistic basis, the availability of a training set and validation. ECOSAR, BIOWIN and DEREK for Windows have a mechanistic or empirical basis. ECOSAR has a training set for each QSAR. For half of the structural fragments the number of chemicals in the training set is >4. Based on structural fragments and log Kow, ECOSAR uses linear regression to predict ecotoxicity. Validating ECOSAR for three 'valid' classes results in predictivity of > or = 64%. BIOWIN uses (non-)linear regressions to predict the probability of biodegradability based on fragments and molecular weight. It has a large training set and predicts non-ready biodegradability well. DEREK for Windows predictions are supported by a mechanistic rationale and literature references. The structural alerts in this program have been developed with a training set of positive and negative toxicity data. However, to support the prediction only a limited number of chemicals in the training set is presented to the user. DEREK for Windows predicts effects by 'if-then' reasoning. The program predicts best for mutagenicity and carcinogenicity. Each structural fragment in ECOSAR and DEREK for Windows needs to be evaluated and validated separately.

Identification of a New DNA Region Specific for Members of Mycobacterium tuberculosis Complex

PubMed Central

Magdalena, Juana; Vachée, Anne; Supply, Philip; Locht, Camille

1998-01-01

The successful use of DNA amplification for the detection of tuberculous mycobacteria crucially depends on the choice of the target sequence, which ideally should be present in all tuberculous mycobacteria and absent from all other bacteria. In the present study we developed a PCR procedure based on the intergenic region (IR) separating two genes encoding a recently identified mycobacterial two-component system named SenX3-RegX3. The senX3-regX3 IR is composed of a novel type of repetitive sequence, called mycobacterial interspersed repetitive units (MIRUs). In a survey of 116 Mycobacterium tuberculosis strains characterized by different IS6110 restriction fragment length polymorphisms, 2 Mycobacterium africanum strains, 3 Mycobacterium bovis strains (including 2 BCG strains), and 1 Mycobacterium microti strain, a specific PCR fragment was amplified in all cases. This collection included M. tuberculosis strains that lack IS6110 or mtp40, two target sequences that have previously been used for the detection of M. tuberculosis. No PCR fragment was amplified when DNA from other organisms was used, giving a sensitivity of 100% and a specificity of 100% in the confidence limit of this study. The numbers of MIRUs were found to vary among strains, resulting in six different groups of strains on the basis of the size of the amplified PCR fragment. However, the vast majority of the strains (approximately 90%) fell within the same group, containing two 77-bp MIRUs followed by one 53-bp MIRU. PMID:9542912

A recombinant estrogen receptor fragment-based homogeneous fluorescent assay for rapid detection of estrogens.

PubMed

Wang, Dan; Xie, Jiangbi; Zhu, Xiaocui; Li, Jinqiu; Zhao, Dongqin; Zhao, Meiping

2014-05-15

In this work, we demonstrate a novel estrogenic receptor fragment-based homogeneous fluorescent assay which enables rapid and sensitive detection of 17β-estradiol (E2) and other highly potent estrogens. A modified human estrogenic receptor fragment (N-His × 6-hER270-595-C-Strep tag II) has been constructed that contains amino acids 270-595 of wild-type human estrogenic receptor α (hER270-595) and two specific tags (6 × His and Strep tag II) fused to the N and C terminus, respectively. The designed receptor protein fragment could be easily produced by prokaryotic expression with high yield and high purity. The obtained protein exhibits high binding affinity to E2 and the two tags greatly facilitate the application of the recombinant protein. Taking advantage of the unique spectroscopic properties of coumestrol (CS), a fluorescent phytoestrogen, a CS/hER270-595-based fluorescent assay has been developed which can sensitively respond to E2 within 1.0 min with a linear working range from 0.1 to 20 ng/mL and a limit of detection of 0.1 ng/mL. The assay was successfully applied for rapid detection of E2 in the culture medium of rat hippocampal neurons. The method also holds great potential for high-throughput monitoring the variation of estrogen levels in complex biological fluids, which is crucial for investigation of the molecular basis of various estrogen-involved processes. Copyright © 2013 Elsevier B.V. All rights reserved.

New Polymeric Precursors of Silicon Carbide

NASA Technical Reports Server (NTRS)

Litt, M.; Kumar, K.

1987-01-01

Silicon carbide made by pyrolizing polymers. Method conceived for preparation of poly(decamethylcyclohexasilanes) as precursors for preparation of silicon carbide at high yield. Technical potential of polysilanes as precursors of SiC ceramics being explored. Potential limited by intractability of some polysilanes; formation of small, cyclic polycarbosilane fragments during pyrolysis; and overall low char yield and large shrinkage in conversion to ceramics.

Word Problems and Make-Believe: Using Frame Analysis and Ethnomethodology to Explore Aspects of the Culture of Schooling

ERIC Educational Resources Information Center

Benincasa, Luciana

2017-01-01

The paper applies Goffman's frame analysis and ethnomethodology to student performance on mathematical word problems. In educational research, frame analysis has usually been limited to primary frames. Instead, in this paper I focus on the kind of secondary frame that Goffman calls 'utilitarian make-believe'. The data consist of a fragment of…

Predicted long-term effects of group selection on species composition and stand structure in northern hardwood forests

Treesearch

Corey R. Halpin; Craig G. Lorimer; Jacob J. Hanson; Brian J. Palik

2017-01-01

The group selection method can potentially increase the proportion of shade-intolerant and midtolerant tree species in forests dominated by shade-tolerant species, but previous results have been variable, and concerns have been raised about possible effects on forest fragmentation and forest structure. Limited evidence is available on these issues for forests managed...

Use of artificial nests to investigate predation on freshwater turtle nests

Treesearch

Michael N. Marchand; John A. Litvaitis; Thomas J. Maier; Richard M. DeGraaf

2002-01-01

Habitat fragmentation has raised concerns that populations of generalist predators have increased and are affecting a diverse group of prey. Previous research has included the use of artificial nests to investigate the role of predation on birds that nest on or near the ground. Because predation also is a major factor limiting populations of freshwater turtles, we...

Using DNase Hi-C techniques to map global and local three-dimensional genome architecture at high resolution.

PubMed

Ma, Wenxiu; Ay, Ferhat; Lee, Choli; Gulsoy, Gunhan; Deng, Xinxian; Cook, Savannah; Hesson, Jennifer; Cavanaugh, Christopher; Ware, Carol B; Krumm, Anton; Shendure, Jay; Blau, C Anthony; Disteche, Christine M; Noble, William S; Duan, ZhiJun

2018-06-01

The folding and three-dimensional (3D) organization of chromatin in the nucleus critically impacts genome function. The past decade has witnessed rapid advances in genomic tools for delineating 3D genome architecture. Among them, chromosome conformation capture (3C)-based methods such as Hi-C are the most widely used techniques for mapping chromatin interactions. However, traditional Hi-C protocols rely on restriction enzymes (REs) to fragment chromatin and are therefore limited in resolution. We recently developed DNase Hi-C for mapping 3D genome organization, which uses DNase I for chromatin fragmentation. DNase Hi-C overcomes RE-related limitations associated with traditional Hi-C methods, leading to improved methodological resolution. Furthermore, combining this method with DNA capture technology provides a high-throughput approach (targeted DNase Hi-C) that allows for mapping fine-scale chromatin architecture at exceptionally high resolution. Hence, targeted DNase Hi-C will be valuable for delineating the physical landscapes of cis-regulatory networks that control gene expression and for characterizing phenotype-associated chromatin 3D signatures. Here, we provide a detailed description of method design and step-by-step working protocols for these two methods. Copyright © 2018 Elsevier Inc. All rights reserved.

Multiframe digitization of x-ray (TV) images (abstract)

NASA Astrophysics Data System (ADS)

Karpenko, V. A.; Khil'chenko, A. D.; Lysenko, A. P.; Panchenko, V. E.

1989-07-01

The work in progress deals with the experimental search for a technique of digitizing x-ray TV images. The small volume of the buffer memory of the analog-to-digital (A/D) converter (ADC) we have previously used to detect TV signals made it necessary to digitize only one line at a time of the television raster and also to make use of gating to gain the video information contained in the whole frame. This paper is devoted to multiframe digitizing. The recorder of video signals comprises a broadband 8-bit A/D converter, a buffer memory having 128K words and a control circuit which forms a necessary sequence of advance pulses for the A/D converter and the memory relative to the input frame and line sync pulses (FSP and LSP). The device provides recording of video signals corresponding to one or a few frames following one after another, or to their fragments. The control circuit is responsible for the separation of the required fragment of the TV image. When loading the limit registers, the following input parameters of the control circuit are set: the skipping of a definite number of lines after the next FSP, the number of the lines of recording inside a fragment, the frequency of the information lines inside a fragment, the delay in the start of the ADC conversion relative to the arrival of the LSP, the length of the information section of a line, and the frequency of taking the readouts in a line. In addition, among the instructions given are the number of frames of recording and the frequency of their sequence. Thus, the A/D converter operates only inside a given fragment of the TV image. The information is introduced into the memory in sequence, fragment by fragment, without skipping and is then extracted as samples according to the addresses needed for representation in the required form, and processing. The video signal recorder governs the shortest time of the ADC conversion per point of 250 ns. As before, among the apparatus used were an image vidicon with luminophor conversion of x-radiation to light, and a single-crystal x-ray diffraction scheme necessary to form dynamic test objects from x-ray lines dispersed in space (the projections of the linear focus of an x-ray tube).

Erosion in extruder flow

NASA Astrophysics Data System (ADS)

Kaufman, Miron; Fodor, Petru S.

A detailed analysis of the fluid flow in Tadmor's unwound channel model of the single screw extruder is performed by combining numerical and analytical methods. Using the analytical solution for the longitudinal velocity field (in the limit of zero Reynolds number) allows us to devote all the computational resources solely for a detailed numerical solution of the transversal velocity field. This high resolution 3D model of the fluid flow in a single-screw extruder allows us to identify the position and extent of Moffatt eddies that impede mixing. We further consider the erosion of particles (e.g. carbon-black agglomerates) advected by the polymeric flow. We assume a particle to be made of primary fragments bound together. In the erosion process a primary fragment breaks out of a given particle. Particles are advected by the laminar flow and they disperse because of the shear stresses imparted by the fluid. The time evolution of the numbers of particles of different sizes is described by the Bateman coupled differential equations used to model radioactivity. Using the particle size distribution we compute an entropic fragmentation index which varies from 0 for a monodisperse system to 1 for an extreme poly-disperse system.

Comparative gene expression in sexual and apomictic ovaries of Pennisetum ciliare (L.) Link.

PubMed

Vielle-Calzada, J P; Nuccio, M L; Budiman, M A; Thomas, T L; Burson, B L; Hussey, M A; Wing, R A

1996-12-01

Limited emphasis has been given to the molecular study of apomixis, an asexual method of reproduction where seeds are produced without fertilization. Most buffelgrass (Pennisetum ciliare (L.) Link syn = Cenchrus ciliaris L.) genotypes reproduce by obligate apomixis (apospory); however, rare sexual plants have been recovered. A modified differential display procedure was used to compare gene expression in unpollinated ovaries containing ovules with either sexual or apomictic female gametophytes. The modification incorporated end-labeled poly(A)+ anchored primers as the only isotopic source, and was a reliable and consistent approach for detecting differentially displayed transcripts. Using 20 different decamers and two anchor primers, 2268 cDNA fragments between 200 and 600 bp were displayed. From these, eight reproducible differentially displayed cDNAs were identified and cloned. Based on northern analysis, one cDNA was detected in only the sexual ovaries, two cDNAs in only apomictic ovaries and one cDNA was present in both types of ovaries. Three fragments could not be detected and one fragment was detected in ovaries, stems, and leaves. Comparison of gene expression during sexual and apomictic development in buffelgrass represents a new model system and a strategy for investigating female reproductive development in the angiosperms.

Quantitation of lysergic acid diethylamide in urine using atmospheric pressure matrix-assisted laser desorption/ionization ion trap mass spectrometry.

PubMed

Cui, Meng; McCooeye, Margaret A; Fraser, Catharine; Mester, Zoltán

2004-12-01

A quantitative method was developed for analysis of lysergic acid diethylamide (LSD) in urine using atmospheric pressure matrix-assisted laser desorption/ionization ion trap mass spectrometry (AP MALDI-ITMS). Following solid-phase extraction of LSD from urine samples, extracts were analyzed by AP MALDI-ITMS. The identity of LSD was confirmed by fragmentation of the [M + H](+) ion using tandem mass spectrometry. The quantification of LSD was achieved using stable-isotope-labeled LSD (LSD-d(3)) as the internal standard. The [M + H](+) ion fragmented to produce a dominant fragment ion, which was used for a selected reaction monitoring (SRM) method for quantitative analysis of LSD. SRM was compared with selected ion monitoring and produced a wider linear range and lower limit of quantification. For SRM analysis of samples of LSD spiked in urine, the calibration curve was linear in the range of 1-100 ng/mL with a coefficient of determination, r(2), of 0.9917. This assay was used to determine LSD in urine samples and the AP MALDI-MS results were comparable to the HPLC/ ESI-MS results.

Conservative management of retained cardiac missiles: case report and literature review.

PubMed

Lundy, Jonathan B; Johnson, Eric K; Seery, Jason M; Pham, Tach; Frizzi, James D; Chasen, Arthur B

2009-01-01

Intracardiac foreign bodies may be caused by direct penetrating trauma, embolization from injury to another area of the body, or iatrogenically from fragments of intravascular access devices. Penetrating cardiac trauma commonly presents with a hemodynamically unstable patient necessitating emergent life-saving procedures. Missile embolization to the heart can occur after injury to systemic and pulmonary veins. Central venous access devices may fracture after placement and embolize. Especially in the setting of penetrating cardiac trauma, these intracardiac foreign bodies require expeditious removal. Limited data exist regarding the conservative management of intracardiac material after trauma. We present the case of a 42-year-old male soldier injured in a mortar blast in Iraq who suffered multiple injuries to include a right hemopneumothorax and soft tissue injuries to the chest and both lower extremities that was found to have a 2-cm by 2-mm intracardiac metal fragment. Additional imaging revealed a metallic fragment localized to the interatrial septum. The patient suffered no adverse sequelae from nonoperative management. A review of the world literature regarding the subject of posttraumatic retained cardiac missiles (RCMs) is also included to help future surgeons in the management of this rare entity.

Identification of Relevant Conformational Epitopes on the HER2 Oncoprotein by Using Large Fragment Phage Display (LFPD)

PubMed Central

Gabrielli, Federico; Salvi, Roberto; Garulli, Chiara; Kalogris, Cristina; Arima, Serena; Tardella, Luca; Monaci, Paolo; Pupa, Serenella M.; Tagliabue, Elda; Montani, Maura; Quaglino, Elena; Stramucci, Lorenzo; Curcio, Claudia

2013-01-01

We developed a new phage-display based approach, the Large Fragment Phage Display (LFPD), that can be used for mapping conformational epitopes on target molecules of immunological interest. LFPD uses a simplified and more effective phage-display approach in which only a limited set of larger fragments (about 100 aa in length) are expressed on the phage surface. Using the human HER2 oncoprotein as a target, we identified novel B-cell conformational epitopes. The same homologous epitopes were also detected in rat HER2 and all corresponded to the epitopes predicted by computational analysis (PEPITO software), showing that LFPD gives reproducible and accurate results. Interestingly, these newly identified HER2 epitopes seem to be crucial for an effective immune response against HER2-overexpressing breast cancers and might help discriminating between metastatic breast cancer and early breast cancer patients. Overall, the results obtained in this study demonstrated the utility of LFPD and its potential application to the detection of conformational epitopes on many other molecules of interest, as well as, the development of new and potentially more effective B-cell conformational epitopes based vaccines. PMID:23555577

Regulation of Cre recombinase by ligand-induced complementation of inactive fragments.

PubMed

Jullien, Nicolas; Sampieri, François; Enjalbert, Alain; Herman, Jean-Paul

2003-11-01

Cre recombinase is extensively used to engineer the genome of experimental animals. However, its usefulness is still limited by the lack of an efficient temporal control over its activity. To overcome this, we have developed DiCre, a regulatable fragment complementation system for Cre. The enzyme was split into two moieties that were fused to FKBP12 (FK506-binding protein) and FRB (binding domain of the FKBP12-rapamycin-associated protein), respectively. These can be efficiently heterodimerized by rapamycin. Several variants, based on splitting Cre at different sites and using different linker peptides, were tested in an indicator cell line. The fusion proteins, taken separately, had no recombinase activity. Stable transformants, co-expressing complementing fragments based on splitting Cre between Asn59 and Asn60, displayed low background activity affecting 0.05-0.4% of the cells. Rapamycin induced a rapid recombination, reaching 100% by 48-72 h, with an EC50 of 0.02 nM. Thus, ligand-induced dimerization can efficiently regulate Cre, and should be useful to achieve a tight temporal control of its activity, such as in the case of the creation of conditional knock-out animals.

Identification of IgE- binding pollen protein from Cannabis sativa in pollen-hypersensitive patients from north Pakistan.

PubMed

Choudhary, Shazia; Murad, Sheeba; Hayat, Muhammad Qasim; Shakoor, Zahid; Arshad, Muhammad

2017-01-01

Cannabis sativa (C.sativa) is well-known for its medicinal, industrial and recreational use. However, allergies in relation to Cannabis sativa (C.sativa) are rarely reported. C. sativa is one of the common weeds found in Pakistan and its pollen grains are common in spring and fall season. Although categorized as an aeroallergen, there are limited number of reports regarding allergenic potential in C. sativa. Therefore, the current study is aimed at exploring the IgE- binding potential among the C. sativa pollen in local pollen allergic patients. Initial screening of C. sativa sensitized individuals was carried out by dot blot from the sera of pollen allergic patients. Proteins from the pollen grains were extracted and resolved on 10% gel. Eight bands were visible on gel however only one protein fragment i.e. of 14KDa size was found to bind to IgE as analyzed through protein gel blot analysis. Strong IgE affinity of a 14 kDa protein fragment from C. sativa pollen extract suggests its allergenic potential. Further study is required to find the exact nature of this protein fragment.

Construction of a food-grade cloning vector for Lactobacillus plantarum and its utilization in a food model.

PubMed

Rattanachaikunsopon, Pongsak; Phumkhachorn, Parichat

2012-01-01

The development of Lactobacillus plantarum to be used in starter cultures in the food industry has been limited because of the lack of a food-grade cloning vector for the bacterium. In this study, the plasmid pFLP1 was constructed by joining 2 DNA fragments derived from food-approved organisms. The 5.2-kb BamHI/KpnI DNA fragment of pRV566 containing the theta-type replicon of Lactobacillus sakei was ligated to the BamHI/KpnI DNA fragment of a 2.9-kb lactococcal cadmium resistance determinant amplified from pND918. The 8.1-kb newly constructed plasmid could transform L. plantarum N014, a bacteriocin-producing bacteria originally isolated from nham, a traditional Thai fermented sausage. The resulting transformant, L. plantarum N014-FLP, and its parent strain were shown to be very similar in growth rate and bacteriocin activity. In addition, the plasmid was very stable in its host bacteria under nonselective pressure for 100 generations in MRS medium and for 5 days in a nham model. These results suggest that pFLP1 is a potential food-grade cloning vector for L. plantarum.

Erosion of ejecta at Meteor Crater, Arizona

NASA Technical Reports Server (NTRS)

Grant, John A.; Schultz, Peter H.

1993-01-01

New methods for estimating erosion at Meteor Crater, Arizona, indicate that continuous ejecta deposits beyond 1/4-1/2 crater radii from the rim have been lowered less than 1 m on the average. This conclusion is based on the results of two approaches: coarsening of unweathered ejecta into surface lag deposits and calculation of the sediment budget within a drainage basin on the ejecta. Preserved ejecta morphologies beneath thin alluvium revealed by ground-penetrating radar provide qualitative support for the derived estimates. Although slightly greater erosion of less resistant ejecta locally has occurred, such deposits were limited in extent, particularly beyond 0.25R-0.5R from the present rim. Subtle but preserved primary ejecta features further support our estimate of minimal erosion of ejecta since the crater formed about 50,000 years ago. Unconsolidated deposits formed during other sudden extreme events exhibit similarly low erosion over the same time frame; the common factor is the presence of large fragments or large fragments in a matrix of finer debris. At Meteor Crater, fluvial and eolian processes remove surrounding fines leaving behind a surface lag of coarse-grained ejecta fragments that armor surfaces and slow vertical lowering.

The movement ecology and dynamics of plant communities in fragmented landscapes

DOE PAGES

Damschen, Ellen I.; Brudvig, Lars A.; Haddad, Nick M.; ...

2008-12-05

A conceptual model of movement ecology has recently been advanced to explain all movement by considering the interaction of four elements: internal state, motion capacity, navigation capacities,and external factors. We modified this framework togenerate predictions for species richness dynamics of fragmented plant communities and tested them in experimental landscapes across a 7-year time series. We found that two external factors, dispersal vectors and habitat features, affected species colonization and recolonization in habitat fragments and their effects varied and depended on motion capacity. Bird-dispersed species richness showed connectivity effects that reached an asymptote over time, but no edge effects, whereas wind-dispersedmore » species richness showed steadily accumulating edge and connectivity effects, with no indication of an asymptote. Unassisted species also showed increasing differences caused by connectivity over time,whereas edges had no effect. Our limited use of proxies for movement ecology (e.g., dispersal mode as a proxy for motion capacity) resulted in moderate predictive power for communities and, in some cases, highlighted the importance of a more complete understanding of movement ecology for predicting how landscape conservation actions affect plant community dynamics.« less

Isobaric yield ratio difference between the 140 A MeV 58Ni + 9Be and 64Ni +9Be reactions studied by the antisymmetric molecular dynamics model

NASA Astrophysics Data System (ADS)

Qiao, C. Y.; Wei, H. L.; Ma, C. W.; Zhang, Y. L.; Wang, S. S.

2015-07-01

Background: The isobaric yield ratio difference (IBD) method is found to be sensitive to the density difference of neutron-rich nucleus induced reaction around the Fermi energy. Purpose: An investigation is performed to study the IBD results in the transport model. Methods: The antisymmetric molecular dynamics (AMD) model plus the sequential decay model gemini are adopted to simulate the 140 A MeV 58 ,64Ni +9Be reactions. A relative small coalescence radius Rc= 2.5 fm is used for the phase space at t = 500 fm/c to form the hot fragment. Two limitations on the impact parameter (b 1 =0 -2 fm and b 2 =0 -9 fm) are used to study the effect of central collisions in IBD. Results: The isobaric yield ratios (IYRs) for the large-A fragments are found to be suppressed in the symmetric reaction. The IBD results for fragments with neutron excess I = 0 and 1 are obtained. A small difference is found in the IBDs with the b 1 and b 2 limitations in the AMD simulated reactions. The IBD with b 1 and b 2 are quite similar in the AMD + GEMINI simulated reactions. Conclusions: The IBDs for the I =0 and 1 chains are mainly determined by the central collisions, which reflects the nuclear density in the core region of the reaction system. The increasing part of the IBD distribution is found due to the difference between the densities in the peripheral collisions of the reactions. The sequential decay process influences the IBD results. The AMD + GEMINI simulation can better reproduce the experimental IBDs than the AMD simulation.

A systematic review of military head injuries.

PubMed

Carr, Debra J; Lewis, E; Horsfall, I

2017-02-01

This commissioned review discusses military head injuries caused by non-ballistic impacts, penetrating fragments and bullets (including parts of bullets) and behind helmet blunt trauma (BHBT). A systematic review of the literature was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses method. The openly accessible literature was reviewed to investigate military head injuries and their severity. Fifty-four sources were identified that included pertinent openly accessible information relevant to this topic. Limited injury data exist for non-ballistic head injuries for UK forces, although some international data exist for parachutists. The majority of fatal head injuries are due to projectiles penetrating through the face rather than through the area of the head covered by the helmet. Penetrating head injuries are primarily caused by fragments, but helmets are more commonly perforated by high-energy rifle bullets than by fragments. No reports of a BHBT injury have been located in the literature. The description of body segment varies among articles and this makes comparisons among datasets difficult. There is a lack of detail regarding the precise position and severity of injuries, and long-term outcome for casualties. It is demonstrated that wearing military helmets reduces fatalities on and off the battlefield. The risk of BHBT injuries is widely referred to, but evidence of their occurrence is not provided by the authors that describe the risk of BHBT occurring. Further research into the causes and severity of head injuries would be useful for designers of military helmets and other associated personal protective equipment, particularly as advances in materials technology means lighter, thinner and more protective helmets are achievable. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

Refrigeration and freezing of porcine tissue does not affect the retardation of fragment simulating projectiles.

PubMed

Breeze, J; Carr, D J; Mabbott, A; Beckett, S; Clasper, J C

2015-05-01

Explosively propelled fragments are the most common cause of injury to UK service personnel in modern conflicts. Numerical injury models to simulate such injuries utilise algorithms based upon gelatin and animal tissue testing but data is limited on many fragment simulating projectiles and these simulants cannot represent human anatomy. Testing with post mortem specimens may overcome this limitation but no information exists about how post mortem tissue changes and storage conditions in humans or animals may affect projectile penetration. Two chisel nosed cylinders (0.49 g and 1.10 g) and a 0.51 g (5 mm) sphere were fired into three groups of porcine tissue (fresh, refrigerated and frozen then refrigerated) and compared to 20% gelatin. Depth of projectile penetration was ascertained with the assistance of computed tomography and kinetic energy absorption by tissues measured using Doppler radar and high speed photography. No difference in depth of penetration was found between porcine tissue stored in the different manners compared with 20% gelatin by impact velocities less than 100 m/s. Insufficient numbers of projectiles were retained in tissue at higher velocities for statistical analysis to be undertaken. Energy absorbed per millimetre of tissue ranged between 0.42 and 0.98 J/mm for different porcine tissue despite differing storage. This pilot study would suggest that the effect of refrigerating or freezing porcine tissue followed by thawing has no effect on its ability to retard these projectiles. Further research is required to ascertain if these results occur at greater velocities and for other types of projectile. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

Investigation of degradation processes in IgG1 monoclonal antibodies by limited proteolysis coupled with weak cation-exchange HPLC.

PubMed

Lau, Hollis; Pace, Danielle; Yan, Boxu; McGrath, Theresa; Smallwood, Scott; Patel, Ketaki; Park, Jihea; Park, Sungae S; Latypov, Ramil F

2010-04-01

A new cation-exchange high-performance liquid chromatography (HPLC) method that separates fragment antigen-binding (Fab) and fragment crystallizable (Fc) domains generated by the limited proteolysis of monoclonal antibodies (mAbs) was developed. This assay has proven to be suitable for studying complex degradation processes involving various immunoglobulin G1 (IgG1) molecules. Assignment of covalent degradations to specific regions of mAbs was facilitated by using Lys-C and papain to generate Fab and Fc fragments with unique, protease-dependent elution times. In particular, this method was useful for characterizing protein variants formed in the presence of salt under accelerated storage conditions. Two isoforms that accumulated during storage were readily identified as Fab-related species prior to mass-spectrometric analysis. Both showed reduced biological activity likely resulting from modifications within or in proximity of the complementarity-determining regions (CDRs). Utility of this assay was further illustrated in the work to characterize light-induced degradations in mAb formulations. In this case, a previously unknown Fab-related species which populated upon light exposure was observed. This species was well resolved from unmodified Fab, allowing for direct and high-purity fractionation. Mass-spectrometric analysis subsequently identified a histidine-related degradation product associated with the CDR2 of the heavy chain. In addition, the method was applied to assess the structural organization of a noncovalent IgG1 dimer. A new species corresponding to a Fab-Fab complex was found, implying that interactions between Fab domains were responsible for dimerization. Overall, the data presented demonstrate the suitability of this cation-exchange HPLC method for studying a wide range of covalent and noncovalent degradations in IgG1 mAbs. 2010 Elsevier B.V. All rights reserved.

DNA Double Strand Break Response and Limited Repair Capacity in Mouse Elongated Spermatids.

PubMed

Ahmed, Emad A; Scherthan, Harry; de Rooij, Dirk G

2015-12-16

Spermatids are extremely sensitive to genotoxic exposures since during spermiogenesis only error-prone non homologous end joining (NHEJ) repair pathways are available. Hence, genomic damage may accumulate in sperm and be transmitted to the zygote. Indirect, delayed DNA fragmentation and lesions associated with apoptotic-like processes have been observed during spermatid elongation, 27 days after irradiation. The proliferating spermatogonia and early meiotic prophase cells have been suggested to retain a memory of a radiation insult leading later to this delayed fragmentation. Here, we used meiotic spread preparations to localize phosphorylate histone H2 variant (γ-H2AX) foci marking DNA double strand breaks (DSBs) in elongated spermatids. This technique enabled us to determine the background level of DSB foci in elongated spermatids of RAD54/RAD54B double knockout (dko) mice, severe combined immunodeficiency SCID mice, and poly adenosine diphosphate (ADP)-ribose polymerase 1 (PARP1) inhibitor (DPQ)-treated mice to compare them with the appropriate wild type controls. The repair kinetics data and the protein expression patterns observed indicate that the conventional NHEJ repair pathway is not available for elongated spermatids to repair the programmed and the IR-induced DSBs, reflecting the limited repair capacity of these cells. However, although elongated spermatids express the proteins of the alternative NHEJ, PARP1-inhibition had no effect on the repair kinetics after IR, suggesting that DNA damage may be passed onto sperm. Finally, our genetic mutant analysis suggests that an incomplete or defective meiotic recombinational repair of Spo11-induced DSBs may lead to a carry-over of the DSB damage or induce a delayed nuclear fragmentation during the sensitive programmed chromatin remodeling occurring in elongated spermatids.

Band-broadening suppressed effect in long turned geometry channel and high-sensitive analysis of DNA sample by using floating electrokinetic supercharging on a microchip.

PubMed

Xu, Zhongqi; Murata, Kenji; Arai, Akihiro; Hirokawa, Takeshi

2010-03-12

A featured microchip owning three big reservoirs and long turned geometry channel was designed to improve the detection limit of DNA fragments by using floating electrokinetic supercharging (FEKS) method. The novel design matches the FEKS preconcentration needs of a large sample volume introduction with electrokinetic injection (EKI), as well as long duration of isotachophoresis (ITP) process to enrich low concentration sample. In the curved channel [ approximately 45.6 mm long between port 1 (P1) and the intersection point of two channels], EKI and ITP were performed while the side port 3 (P3) was electrically floated. The turn-induced band broadening with or without ITP process was investigated by a computer simulation (using CFD-ACE+ software) when the analytes traveling through the U-shaped geometry. It was found that the channel curvature determined the extent of band broadening, however, which could be effectively eliminated by the way of ITP. After the ITP-stacked zones passed the intersection point from P1, they were rapidly destacked for separation and detection from ITP to zone electrophoresis by using leading ions from P3. The FEKS carried on the novel chip successfully contributed to higher sensitivities of DNA fragments in comparison with our previous results realized on either a single channel or a cross microchip. The analysis of low concentration 50 bp DNA step ladders (0.23 mugml after 1500-fold diluted) was achieved with normal UV detection at 260 nm. The obtained limit of detections (LODs) were on average 100 times better than using conventional pinched injection, down to several ngml for individual DNA fragment.

Traceability of plant contribution in olive oil by amplified fragment length polymorphisms.

PubMed

Pafundo, Simona; Agrimonti, Caterina; Marmiroli, Nelson

2005-09-07

Application of DNA molecular markers to traceability of foods is thought to bring new benefit to consumer's protection. Even in a complex matrix such as olive oil, DNA could be traced with PCR markers such as the amplified fragment length polymorphisms (AFLPs). In this work, fluorescent AFLPs were optimized for the characterization of olive oil DNA, to obtain highly reproducible, high-quality fingerprints, testing different parameters: the concentrations of dNTPs and labeled primer, the kind of Taq DNA polymerase and thermal cycler, and the quantity of DNA employed. It was found that correspondence of fingerprinting by comparing results in oils and in plants was close to 70% and that the DNA extraction from olive oil was the limiting step for the reliability of AFLP profiles, due to the complex matrix analyzed.

Isolator fragmentation and explosive initiation tests

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dickson, Peter; Rae, Philip John; Foley, Timothy J.

2016-09-19

Three tests were conducted to evaluate the effects of firing an isolator in proximity to a barrier or explosive charge. The tests with explosive were conducted without a barrier, on the basis that since any barrier will reduce the shock transmitted to the explosive, bare explosive represents the worst-case from an inadvertent initiation perspective. No reaction was observed. The shock caused by the impact of a representative plastic material on both bare and cased PBX 9501 is calculated in the worst-case, 1-D limit, and the known shock response of the HE is used to estimate minimum run-to-detonation lengths. The estimatesmore » demonstrate that even 1-D impacts would not be of concern and that, accordingly, the divergent shocks due to isolator fragment impact are of no concern as initiating stimuli.« less

Isolator fragmentation and explosive initiation tests

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dickson, Peter; Rae, Philip John; Foley, Timothy J.

2015-09-30

Three tests were conducted to evaluate the effects of firing an isolator in proximity to a barrier or explosive charge. The tests with explosive were conducted without barrier, on the basis that since any barrier will reduce the shock transmitted to the explosive, bare explosive represents the worst-case from an inadvertent initiation perspective. No reaction was observed. The shock caused by the impact of a representative plastic material on both bare and cased PBX9501 is calculated in the worst-case, 1-D limit, and the known shock response of the HE is used to estimate minimum run-to-detonation lengths. The estimates demonstrate thatmore » even 1-D impacts would not be of concern and that, accordingly, the divergent shocks due to isolator fragment impact are of no concern as initiating stimuli.« less

An efficient Cellular Potts Model algorithm that forbids cell fragmentation

NASA Astrophysics Data System (ADS)

Durand, Marc; Guesnet, Etienne

2016-11-01

The Cellular Potts Model (CPM) is a lattice based modeling technique which is widely used for simulating cellular patterns such as foams or biological tissues. Despite its realism and generality, the standard Monte Carlo algorithm used in the scientific literature to evolve this model preserves connectivity of cells on a limited range of simulation temperature only. We present a new algorithm in which cell fragmentation is forbidden for all simulation temperatures. This allows to significantly enhance realism of the simulated patterns. It also increases the computational efficiency compared with the standard CPM algorithm even at same simulation temperature, thanks to the time spared in not doing unrealistic moves. Moreover, our algorithm restores the detailed balance equation, ensuring that the long-term stage is independent of the chosen acceptance rate and chosen path in the temperature space.

FlavonoidSearch: A system for comprehensive flavonoid annotation by mass spectrometry.

PubMed

Akimoto, Nayumi; Ara, Takeshi; Nakajima, Daisuke; Suda, Kunihiro; Ikeda, Chiaki; Takahashi, Shingo; Muneto, Reiko; Yamada, Manabu; Suzuki, Hideyuki; Shibata, Daisuke; Sakurai, Nozomu

2017-04-28

Currently, in mass spectrometry-based metabolomics, limited reference mass spectra are available for flavonoid identification. In the present study, a database of probable mass fragments for 6,867 known flavonoids (FsDatabase) was manually constructed based on new structure- and fragmentation-related rules using new heuristics to overcome flavonoid complexity. We developed the FlavonoidSearch system for flavonoid annotation, which consists of the FsDatabase and a computational tool (FsTool) to automatically search the FsDatabase using the mass spectra of metabolite peaks as queries. This system showed the highest identification accuracy for the flavonoid aglycone when compared to existing tools and revealed accurate discrimination between the flavonoid aglycone and other compounds. Sixteen new flavonoids were found from parsley, and the diversity of the flavonoid aglycone among different fruits and vegetables was investigated.

Discovery of Fragment-Derived Small Molecules for in Vivo Inhibition of Ketohexokinase (KHK)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Huard, Kim; Ahn, Kay; Amor, Paul

Increased fructose consumption and its subsequent metabolism have been implicated in hepatic steatosis, dyslipidemia, obesity, and insulin resistance in humans. Since ketohexokinase (KHK) is the principal enzyme responsible for fructose metabolism, identification of a selective KHK inhibitor may help to further elucidate the effect of KHK inhibition on these metabolic disorders. Until now, studies on KHK inhibition with small molecules have been limited due to the lack of viable in vivo pharmacological tools. Herein we report the discovery of 12, a selective KHK inhibitor with potency and properties suitable for evaluating KHK inhibition in rat models. Key structural features interactingmore » with KHK were discovered through fragment-based screening and subsequent optimization using structure-based drug design, and parallel medicinal chemistry led to the identification of pyridine 12.« less

Enzyme-Linked Immunosorbent Assay and Serologic Responses to Pneumocystis jiroveci

PubMed Central

Koch, Judy; Levin, Linda; Walzer, Peter D.

2004-01-01

Seroepidemiologic studies of Pneumocystis pneumonia (PCP) in humans have been limited by inadequate reagents. We have developed an enzyme-linked immunosorbent assay (ELISA) using three overlapping recombinant fragments of the human Pneumocystis major surface glycoprotein (MsgA, MsgB, and MsgC) for analysis of antibody responses in HIV-positive patients and healthy blood donors. HIV-positive patients had significantly higher antibody levels to all Msg fragments. Furthermore, HIV-positive patients who experienced a previous episode of PCP (PCP-positive) had higher level of antibodies to MsgC than patients who never had PCP. A significant association was found between ELISA antibody level and reactivity by Western blot in HIV-positive patients, especially those who were PCP-positive. Thus, this ELISA will be useful in studying serum antibody responses to Pneumocystis in different human populations. PMID:15200818

Effect of spatial bias on the nonequilibrium phase transition in a system of coagulating and fragmenting particles.

PubMed

Rajesh, R; Krishnamurthy, Supriya

2002-10-01

We examine the effect of spatial bias on a nonequilibrium system in which masses on a lattice evolve through the elementary moves of diffusion, coagulation, and fragmentation. When there is no preferred directionality in the motion of the masses, the model is known to exhibit a nonequilibrium phase transition between two different types of steady state, in all dimensions. We show analytically that introducing a preferred direction in the motion of the masses inhibits the occurrence of the phase transition in one dimension, in the thermodynamic limit. A finite-size system, however, continues to show a signature of the original transition, and we characterize the finite-size scaling implications of this. Our analysis is supported by numerical simulations. In two dimensions, bias is shown to be irrelevant.

Does avian species richness in natural patch mosaics follow the forest fragmentation paradigm?

USGS Publications Warehouse

Pavlacky, D.C.; Anderson, S.H.

2007-01-01

As one approaches the north-eastern limit of pinyon (Pinus spp.) juniper (Juniperus spp.) vegetation on the Colorado Plateau, USA, woodland patches become increasingly disjunct, grading into sagebrush (Artemisia spp.)-dominated landscapes. Patterns of avian species richness in naturally heterogeneous forests may or may not respond to patch discontinuity in the same manner as bird assemblages in fragmented agricultural systems. We used observational data from naturally patchy woodlands and predictions derived from studies of human-modified agricultural forests to estimate the effects of patch area, shape, isolation and distance to contiguous woodland on avian species richness. We predicted that patterns of species richness in naturally patchy juniper woodlands would differ from those observed in fragmented agricultural systems. Our objectives were to (1) estimate the effect of naturally occurring patch structure on avian species richness with respect to habitat affinity and migratory strategy and (2) assess the concordance of the effects to predictions from agricultural forest systems. We used the analogy between populations and communities to estimate species richness, where species are treated as individuals in the application of traditional capture-recapture theory. Information-theoretic model selection showed that overall species richness was explained primarily by the species area relationship. There was some support for a model with greater complexity than the equilibrium theory of island biogeography where the isolation of large patches resulted in greater species richness. Species richness of woodland-dwelling birds was best explained by the equilibrium hypothesis with partial landscape complementation by open-country species in isolated patches. Species richness within specific migratory strategies showed concomitant increases and no shifts in species composition along the patch area gradient. Our results indicate that many patterns of species richness considered to be fragmentation effects may be general consequences of patch discontinuity and are ubiquitous in naturally heterogeneous systems. There was no evidence for the effects of patch shape and distance to contiguous woodland in juniper woodland, suggesting edge effects and dependence upon regional species pools are characteristics of fragmented agricultural systems. Natural patch mosaics may provide benchmarks for evaluating fragmentation effects and managing forests by mimicking natural landscape patterns. ?? 2007 The Zoological Society of London.

Next generation haplotyping to decipher nuclear genomic interspecific admixture in Citrus species: analysis of chromosome 2.

PubMed

Curk, Franck; Ancillo, Gema; Garcia-Lor, Andres; Luro, François; Perrier, Xavier; Jacquemoud-Collet, Jean-Pierre; Navarro, Luis; Ollitrault, Patrick

2014-12-29

The most economically important Citrus species originated by natural interspecific hybridization between four ancestral taxa (Citrus reticulata, Citrus maxima, Citrus medica, and Citrus micrantha) and from limited subsequent interspecific recombination as a result of apomixis and vegetative propagation. Such reticulate evolution coupled with vegetative propagation results in mosaic genomes with large chromosome fragments from the basic taxa in frequent interspecific heterozygosity. Modern breeding of these species is hampered by their complex heterozygous genomic structures that determine species phenotype and are broken by sexual hybridisation. Nevertheless, a large amount of diversity is present in the citrus gene pool, and breeding to allow inclusion of desirable traits is of paramount importance. However, the efficient mobilization of citrus biodiversity in innovative breeding schemes requires previous understanding of Citrus origins and genomic structures. Haplotyping of multiple gene fragments along the whole genome is a powerful approach to reveal the admixture genomic structure of current species and to resolve the evolutionary history of the gene pools. In this study, the efficiency of parallel sequencing with 454 methodology to decipher the hybrid structure of modern citrus species was assessed by analysis of 16 gene fragments on chromosome 2. 454 amplicon libraries were established using the Fluidigm array system for 48 genotypes and 16 gene fragments from chromosome 2. Haplotypes were established from the reads of each accession and phylogenetic analyses were performed using the haplotypic data for each gene fragment. The length of 454 reads and the level of differentiation between the ancestral taxa of modern citrus allowed efficient haplotype phylogenetic assignations for 12 of the 16 gene fragments. The analysis of the mixed genomic structure of modern species and cultivars (i) revealed C. maxima introgressions in modern mandarins, (ii) was consistent with previous hypotheses regarding the origin of secondary species, and (iii) provided a new picture of the evolution of chromosome 2. 454 sequencing was an efficient strategy to establish haplotypes with significant phylogenetic assignations in Citrus, providing a new picture of the mixed structure on chromosome 2 in 48 citrus genotypes.

Fragmentation and growth processes in ultramafic pseudotachylites and associated wall rocks from Alpine Corsica

NASA Astrophysics Data System (ADS)

Dunkel, K.; Austrheim, H.; Plümper, O.; Ildefonse, B.; Ohl, M.; Jamtveit, B.

2017-12-01

Pseudotachylites, often taken as indicators of seismic slip, can form by comminution and/or melting. How the seismic energy released during a pseudotachylite-generating earthquake affects the wall rock surrounding the fault plane is not well known, although damage of the wall rock can have a substantial influence on the rheological properties of the rock and therefore on its further deformation and metamorphism. Micro- and nanostructures of pseudotachylites and their wall rocks in peridotites from Alpine Corsica have been characterized using optical, scanning and transmission electron microscopy, electron backscatter diffraction, and electron microprobe analysis. Pseudotachylites in dunites from the Cima di Gratera area in Cape Corse show a combination of comminution and melting. Olivine in the pseudotachylites is dislocation-rich whereas pyroxene is almost free of dislocations, indicating a fragmentation of the olivine during seismic slip and incomplete melting. The temperature pulse that induced melting was likely short-lived, since evidence for recovery in olivine is limited. Olivines in the wall rocks show a similarly high dislocation density and are strongly fragmented without being sheared. A fragmentation without shear strain, comparable to the "pulverization" described from shallower earthquakes, can probably only be explained by a high stress pulse released during a seismic event. Previous observations from the same area describe a transition from cataclastic wall rocks over mylonites and ultramylonites to pseudotachylite. This was taken as an indication that cataclasis weakened the rock and finally, in connection with thermal runaway, allowed for seismic slip. This difference between previous observations and those described here may be explained by a spatial heterogeneity in the deformation conditions. While earthquakes may have nucleated by self-localizing thermal runaway in cataclastic zones, they propagated also through previously undeformed rocks, causing fragmentation and partial melting at high stress. The fragmentation of the wall rock, whether it occurred by brittle or crystal-plastic mechanisms, allowed for the infiltration of fluids. Thus, earthquakes cause not only a physical, but also a chemical alteration of the wall rocks surrounding the fault plane

The Disruption and Demise of Periodic Comet Shoemaker-Levy 9

NASA Technical Reports Server (NTRS)

Asphaug, Erik; Benz, Willy; Cuzzi, Jeffrey (Technical Monitor)

1994-01-01

The impact of the fragmented comet Shoemaker-Levy 9 (SL9) into Jupiter this July promises to change our understanding of the outer solar system. More than twenty mountain-sized conglomerates of ice and rock will hit the atmosphere at approx. 50 km/s over the course of a week beginning July 16, releasing approx. 10(exp 4) to 10(exp8) megatons of energy per burst, and providing unique and perhaps pivotal clues to the properties of comets and the physics of massive atmospheres. Because the fragments will strike the far side of Jupiter, data acquisition, analysis and interpretation will be quite sensitive to the actual size and energy of the fragments. We therefore examine an event which took place two summers ago, unnoticed and unobserved: the disruption of SL9 into a "string of pearls' as it passed within the Roche limit at perijove. We first demonstrate, on the basis of timescales of tidal interaction, that the comet could not have broken into 20+ fragments through a hierarchy of brittle fracture events. Next, noting that the tidal stress was too weak to have even fragmented an uncompressed mass of freshly fallen snow, we run models for a strengthless comet held together only by self-gravity. We explore the initial size, density, and rotation. We conclude that a 4 km diameter comet (smaller if a prograde rotator) of density approx. 0.5 g/cu cm disrupts and disperses into a chain of fragments similar to Shoemaker-Levy 9, whether we begin with 21, 85, 169, 700 or 2000 sub-grains. Gravitational reaccumulation is evidently the answer, and there is no need to invoke the presence of 21 "cometesimals" as the subscale of the comet. To explain how a comet can be weaker than uncompacted snow, we show that the ring-plane crossing prior to perijove could have caused total damage. Finally, we compute the tidal stress on impactors as they approach Jupiter this July. Objects of various density are moderately distorted but not disrupted by the time they strike the planet.

A framework to optimize the restoration and retention of large mature forest tracts in managed boreal landscapes.

PubMed

Bouchard, Mathieu; Garet, Jérôme

The decreasing abundance of mature forests and their fragmentation have been identified as major threats for the preservation of biodiversity in managed landscapes. In this study, we developed a multi-level framework to coordinate forest harvestings so as to optimize the retention or restoration of large mature forest tracts in managed forests. We used mixed-integer programming for this optimization, and integrated realistic management assumptions regarding stand yield and operational harvest constraints. The model was parameterized for eastern Canadian boreal forests, where clear-cutting is the main silvicultural system, and is used to examine two hypotheses. First, we tested if mature forest tract targets had more negative impacts on wood supplies when implemented in landscapes that are very different from targeted conditions. Second, we tested the hypothesis that using more partial cuts can be useful to attenuate the negative impacts of mature forest targets on wood supplies. The results indicate that without the integration of an explicit mature forest tract target, the optimization leads to relatively high fragmentation levels. Forcing the retention or restoration of large mature forest tracts on 40% of the landscapes had negative impacts on wood supplies in all types of landscapes, but these impacts were less important in landscapes that were initially fragmented. This counter-intuitive result is explained by the presence in the models of an operational constraint that forbids diffuse patterns of harvestings, which are more costly. Once this constraint is applied, the residual impact of the mature forest tract target is low. The results also indicate that partial cuts are of very limited use to attenuate the impacts of mature forest tract targets on wood supplies in highly fragmented landscapes. Partial cuts are somewhat more useful in landscapes that are less fragmented, but they have to be well coordinated with clearcut schedules in order to contribute efficiently to conservation objectives. This modeling framework could easily be adapted and parameterized to test hypotheses or to optimize restoration schedules in landscapes where issues such as forest fragmentation and the abundance of mature or old-growth forests are a concern.

Formation of a1 Ions Directly from Oxazolone b2 Ions: an Energy-Resolved and Computational Study

NASA Astrophysics Data System (ADS)

Bythell, Benjamin J.; Harrison, Alex G.

2015-05-01

It is well-known that oxazolone b2 ions fragment extensively by elimination of CO to form a2 ions, which often fragment further to form a1 ions. Less well-known is that some oxazolone b2 ions may fragment directly to form a1 ions. The present study uses energy-resolved collision-induced dissociation experiments to explore the occurrence of the direct b2→a1 fragmentation reaction. The experimental results show that the direct b2→a1 reaction is generally observed when Gly is the C-terminal residue of the oxazolone. When the C-terminal residue is more complex, it is able to provide increased stability of the a2 product in the b2→a2 fragmentation pathway. Our computational studies of the relative critical reaction energies for the b2→a2 reaction compared with those for the b2→a1 reaction provide support that the critical reaction energies are similar for the two pathways when the C-terminal residue of the oxazolone is Gly. By contrast, when the nitrogen of the oxazolone ring in the b2 ion does not bear a hydrogen, as in the Ala-Sar and Tyr-Sar (Sar = N-methylglycine) oxazolone b2 ions, a1 ions are not formed but rather neutral imine elimination from the N-terminus of the b2 ion becomes a dominant fragmentation reaction. The M06-2X/6-31+G(d,p) density functional theory calculations are in general agreement with the experimental data for both types of reaction. In contrast, the B3LYP/6-31+G(d,p) model systematically underestimates the barriers of these SN2-like b2→a1 reaction. The difference between the two methods of barrier calculation are highly significant ( P < 0.001) for the b2→a1 reaction, but only marginally significant ( P = 0.05) for the b2→a2 reaction. The computations provide further evidence of the limitations of the B3LYP functional when describing SN2-like reactions.

Surfactants at the Design Limit.

PubMed

Czajka, Adam; Hazell, Gavin; Eastoe, Julian

2015-08-04

This article analyzes how the individual structural elements of surfactant molecules affect surface properties, in particular, the point of reference defined by the limiting surface tension at the aqueous cmc, γcmc. Particular emphasis is given to how the chemical nature and structure of the hydrophobic tails influence γcmc. By comparing the three different classes of surfactants, fluorocarbon, silicone, and hydrocarbon, a generalized surface packing index is introduced which is independent of the chemical nature of the surfactants. This parameter ϕcmc represents the volume fraction of surfactant chain fragments in a surface film at the aqueous cmc. It is shown that ϕcmc is a useful index for understanding the limiting surface tension of surfactants and can be useful for designing new superefficient surfactants.

Genetic diversity and the mating system of a rare Mexican Piñon, Pinus pinceana, and a comparison with Pinus maximartinezii (Pinaceae)

Treesearch

F. Thomas Ledig; Miguel A. Capó-Arteaga; Paul D. Hodgskiss; Hassan Sbay; Celestino Flores-López; M. Thompson Conkle; Basilio Bermejo-Velázquez

2001-01-01

Weeping piñon (Pinus pinceana) has a restricted and fragmented range, trees are widely scattered within populations, and reproduction is limited. Nevertheless, genetic diversity was high; based on 27 isozyme loci in 18 enzyme systems, unbiased expected heterozygosity averaged 0.174. Differentiation also was high (FST = 0.152),...

Enhanced Vaccine Control of Epidemics in Adaptive Networks

DTIC Science & Technology

2010-04-29

than random vaccination 32. When vaccine is very limited, outbreaks can be minimized by fragmenting the network via a graph partitioning strategy...Although an outbreak could transiently decrease network connectivity in a real social network, long term reductions in average connectivity are...37, cholera and typhoid fever 35, as well as pertussis 38. Although not all of the above diseases currently possess a vaccine, research is ongo

Assessing multi-taxa sensitivity to the human footprint, habitat fragmentation and loss by exploring alternative scenarios of dispersal ability and population size: A simulation approach

Treesearch

Brian K. Hand; Samuel A. Cushman; Erin L. Landguth; John Lucotch

2014-01-01

Quantifying the effects of landscape change on population connectivity is compounded by uncertainties about population size and distribution and a limited understanding of dispersal ability for most species. In addition, the effects of anthropogenic landscape change and sensitivity to regional climatic conditions interact to strongly affect habitat...

Predicting the velocity and azimuth of fragments generated by the range destruction or random failure of rocket casings and tankage

NASA Astrophysics Data System (ADS)

Eck, M.; Mukunda, M.

The proliferation of space vehicle launch sites and the projected utilization of these facilities portends an increase in the number of on-pad, ascent, and on-orbit solid-rocket motor (SRM) casings and liquid-rocket tanks which will randomly fail or will fail from range destruct actions. Beyond the obvious safety implications, these failures may have serious resource implications for mission system and facility planners. SRM-casing failures and liquid-rocket tankage failures result in the generation of large, high velocity fragments which may be serious threats to the safety of launch support personnel if proper bunkers and exclusion areas are not provided. In addition, these fragments may be indirect threats to the general public's safety if they encounter hazardous spacecraft payloads which have not been designed to withstand shrapnel of this caliber. They may also become threats to other spacecraft if, by failing on-orbit, they add to the ever increasing space-junk collision cross-section. Most prior attempts to assess the velocity of fragments from failed SRM casings have simply assigned the available chamber impulse to available casing and fuel mass and solved the resulting momentum balance for velocity. This method may predict a fragment velocity which is high or low by a factor of two depending on the ratio of fuel to casing mass extant at the time of failure. Recognizing the limitations of existing methods, the authors devised an analytical approach which properly partitions the available impulse to each major system-mass component. This approach uses the Physics International developed PISCES code to couple the forces generated by an Eulerian modeled gas flow field to a Lagrangian modeled fuel and casing system. The details of a predictive analytical modeling process as well as the development of normalized relations for momentum partition as a function of SRM burn time and initial geometry are discussed in this paper. Methods for applying similar modeling techniques to liquid-tankage-over-pressure failures are also discussed. These methods have been calibrated against observed SRM ascent failures and on-orbit tankage failures. Casing-quadrant sized fragments with velocities exceeding 100 m/s resulted from Titan 34D-SRM range destruct actions at 10 s mission elapsed time (MET). Casing-quadrant sized fragments with velocities of approx. 200 m/s resulted from STS-SRM range destruct actions at 110 s MET. Similar sized fragments for Ariane third stage and Delta second stage tankage were predicted to have maximum velocities of 260 and 480 m/s respectively. Good agreement was found between the predictions and observations for five specific events and it was concluded that the methods developed have good potential for use in predicting the fragmentation process of a number of generically similar casing and tankage systems.

Storm-generated coral fragments - A viable source of transplants for reef rehabilitation

USGS Publications Warehouse

Garrison, V.; Ward, G.

2008-01-01

Coral reefs throughout the world have been damaged by storms, diseases, coral predators, temperature anomalies, and human activities. During the past three decades, recovery has been limited and patchy. Although a damaged coral reef cannot be restored to its original condition, interest in reef restoration is increasing. In a pilot project in the Caribbean (US Virgin Islands), storm-produced fragments of Acropora palmata, A. cervicornis, and Porites porites were collected from donor reefs and transplanted to nearby degraded reefs. Sixty coral fragments were attached to dead-coral substrate (usually A. palmata skeletons), at similar depths from which they had been collected (1-3.5 m), using nylon cable ties. Seventy-five intact colonies were designated as controls. Study colonies were assessed at 6-month intervals for 2 years (1999-2001) and annually thereafter (through 2004). One-fourth of the 135 colonies and fragments monitored were alive at the conclusion of the 5-year study. Survival of control and transplanted A. cervicornis and P. porites was very low (median survival 2.4 and 1.8 years, respectively), with no significant differences between transplant and control colonies. Site and depth did not contribute significantly to A. palmata colony survival, but colony size and transplant/control status did. Probability of survival increased with colony size. Median survival for A. palmata was 1.3 years for transplant and 4.3 years for natural colonies when not controlled for size. A. palmata was the only viable candidate for reef rehabilitation. Storm swells were the primary cause of mortality.

New method to determine the total carbonyl functional group content in extractable particulate organic matter by tandem mass spectrometry.

PubMed

Dron, J; Zheng, W; Marchand, N; Wortham, H

2008-08-01

A functional group analysis method was developed to determine the quantitative content of carbonyl functional groups in atmospheric particulate organic matter (POM) using constant neutral loss scanning-tandem mass spectrometry (CNLS-MS/MS). The neutral loss method consists in monitoring the loss of a neutral fragment produced by the fragmentation of a precursor ion in a collision cell. The only ions detected are the daughter ions resulting from the loss of the neutral fragment under study. Then, scanning the loss of a neutral fragment characteristic of a functional group enables the selective detection of the compounds bearing the chemical function under study within a complex mixture. The selective detection of carbonyl functional groups was achieved after derivatization with pentafluorophenylhydrazine (PFPH) by monitoring the neutral loss of C(6)F(5)N (181 amu), which was characteristic of a large panel of derivatized carbonyl compounds. The method was tested on 25 reference mixtures of different composition, all containing 24 carbonyl compounds at randomly determined concentrations. The repeatability and calibration tests were satisfying as they resulted in a relative standard deviation below 5% and a linear range between 0.01 and 0.65 mM with a calculated detection limit of 0.0035 mM. Also, the relative deviation induced by changing the composition of the mixture while keeping the total concentration of carbonyl functional groups constant was less than 20%. These reliability experiments demonstrate the high robustness of the developed procedure for accurate carbonyl functional group measurement, which was applied to atmospheric POM samples. Copyright (c) 2008 John Wiley & Sons, Ltd.

Entropy-driven one-step formation of Phi29 pRNA 3WJ from three RNA fragments.

PubMed

Binzel, Daniel W; Khisamutdinov, Emil F; Guo, Peixuan

2014-04-15

The emerging field of RNA nanotechnology necessitates creation of functional RNA nanoparticles but has been limited by particle instability. It has been shown that the three-way junction of bacteriophage phi29 motor pRNA has unusual stability and can self-assemble from three fragments with high efficiency. It is generally believed that RNA and DNA folding is energy landscape-dependent, and the folding of RNA is driven by enthalpy. Here we examine the thermodynamic characteristics of the 3WJ components as 2'-fluoro RNA, DNA, and RNA. It was seen that the three fragments existed either in 3WJ complex or as monomers, with the intermediate of dimers almost undetectable. It seems that the three fragments can lead to the formation of the 3WJ complex efficiently within a rapid time. A low dissociation constant (apparent KD) of 11.4 nM was determined for RNA, inclusion of 2'-F pyrimidines strengthened the KD to 4.5 nM, and substitution of DNA weakened it to 47.7 nM. The ΔG°37, were -36, -28, and -15 kcal/mol for 3WJ2'-F, 3WJRNA, and 3WJDNA, respectively. It is found that the formation of the three-component complex was governed by entropy, instead of enthalpy, as usually found in RNA complexes. Here entropy-driven is referring to a dominating entropic contribution to the increased stability of the 3WJ(2'-F and 3WJ(RNA) compared to the 3WJ(DNA,) instead of referring to the absolute role or total energy governing 3WJ folding. [corrected].

The impact of homologous recombination repair deficiency on depleted uranium clastogenicity in Chinese hamster ovary cells: XRCC3 protects cells from chromosome aberrations, but increases chromosome fragmentation.

PubMed

Holmes, Amie L; Joyce, Kellie; Xie, Hong; Falank, Carolyne; Hinz, John M; Wise, John Pierce

2014-04-01

Depleted uranium (DU) is extensively used in both industry and military applications. The potential for civilian and military personnel exposure to DU is rising, but there are limited data on the potential health hazards of DU exposure. Previous laboratory research indicates DU is a potential carcinogen, but epidemiological studies remain inconclusive. DU is genotoxic, inducing DNA double strand breaks, chromosome damage and mutations, but the mechanisms of genotoxicity or repair pathways involved in protecting cells against DU-induced damage remain unknown. The purpose of this study was to investigate the effects of homologous recombination repair deficiency on DU-induced genotoxicity using RAD51D and XRCC3-deficient Chinese hamster ovary (CHO) cell lines. Cells deficient in XRCC3 (irs1SF) exhibited similar cytotoxicity after DU exposure compared to wild-type (AA8) and XRCC3-complemented (1SFwt8) cells, but DU induced more break-type and fusion-type lesions in XRCC3-deficient cells compared to wild-type and XRCC3-complemented cells. Surprisingly, loss of RAD51D did not affect DU-induced cytotoxicity or genotoxicity. DU induced selective X-chromosome fragmentation irrespective of RAD51D status, but loss of XRCC3 nearly eliminated fragmentation observed after DU exposure in wild-type and XRCC3-complemented cells. Thus, XRCC3, but not RAD51D, protects cells from DU-induced breaks and fusions and also plays a role in DU-induced chromosome fragmentation. Copyright © 2014 Elsevier B.V. All rights reserved.

Sex Determination from Fragmented and Degenerated DNA by Amplified Product-Length Polymorphism Bidirectional SNP Analysis of Amelogenin and SRY Genes.

PubMed

Masuyama, Kotoka; Shojo, Hideki; Nakanishi, Hiroaki; Inokuchi, Shota; Adachi, Noboru

2017-01-01

Sex determination is important in archeology and anthropology for the study of past societies, cultures, and human activities. Sex determination is also one of the most important components of individual identification in criminal investigations. We developed a new method of sex determination by detecting a single-nucleotide polymorphism in the amelogenin gene using amplified product-length polymorphisms in combination with sex-determining region Y analysis. We particularly focused on the most common types of postmortem DNA damage in ancient and forensic samples: fragmentation and nucleotide modification resulting from deamination. Amplicon size was designed to be less than 60 bp to make the method more useful for analyzing degraded DNA samples. All DNA samples collected from eight Japanese individuals (four male, four female) were evaluated correctly using our method. The detection limit for accurate sex determination was determined to be 20 pg of DNA. We compared our new method with commercial short tandem repeat analysis kits using DNA samples artificially fragmented by ultraviolet irradiation. Our novel method was the most robust for highly fragmented DNA samples. To deal with allelic dropout resulting from deamination, we adopted "bidirectional analysis," which analyzed samples from both sense and antisense strands. This new method was applied to 14 Jomon individuals (3500-year-old bone samples) whose sex had been identified morphologically. We could correctly identify the sex of 11 out of 14 individuals. These results show that our method is reliable for the sex determination of highly degenerated samples.

Sex Determination from Fragmented and Degenerated DNA by Amplified Product-Length Polymorphism Bidirectional SNP Analysis of Amelogenin and SRY Genes

PubMed Central

Masuyama, Kotoka; Shojo, Hideki; Nakanishi, Hiroaki; Inokuchi, Shota; Adachi, Noboru

2017-01-01

Sex determination is important in archeology and anthropology for the study of past societies, cultures, and human activities. Sex determination is also one of the most important components of individual identification in criminal investigations. We developed a new method of sex determination by detecting a single-nucleotide polymorphism in the amelogenin gene using amplified product-length polymorphisms in combination with sex-determining region Y analysis. We particularly focused on the most common types of postmortem DNA damage in ancient and forensic samples: fragmentation and nucleotide modification resulting from deamination. Amplicon size was designed to be less than 60 bp to make the method more useful for analyzing degraded DNA samples. All DNA samples collected from eight Japanese individuals (four male, four female) were evaluated correctly using our method. The detection limit for accurate sex determination was determined to be 20 pg of DNA. We compared our new method with commercial short tandem repeat analysis kits using DNA samples artificially fragmented by ultraviolet irradiation. Our novel method was the most robust for highly fragmented DNA samples. To deal with allelic dropout resulting from deamination, we adopted “bidirectional analysis,” which analyzed samples from both sense and antisense strands. This new method was applied to 14 Jomon individuals (3500-year-old bone samples) whose sex had been identified morphologically. We could correctly identify the sex of 11 out of 14 individuals. These results show that our method is reliable for the sex determination of highly degenerated samples. PMID:28052096

Riparian corridors enhance movement of a forest specialist bird in fragmented tropical forest.

PubMed

Gillies, Cameron S; St Clair, Colleen Cassady

2008-12-16

Riparian corridors and fencerows are hypothesized to increase the persistence of forest animals in fragmented landscapes by facilitating movement among suitable habitat patches. This function may be critically important for forest birds, which have declined dramatically in fragmented habitats. Unfortunately, direct evidence of corridor use has been difficult to collect at landscape scales and this limits support for corridors in conservation planning. Using telemetry and handheld GPS units, we examined the movement of forest birds by translocating territorial individuals of barred antshrikes (Thamnophilus doliatus; a forest specialist) and rufous-naped wrens (Campylorhynchus rufinucha; a forest generalist) 0.7-1.9 km from their territories in the highly fragmented tropical dry forest of Costa Rica. In each translocation, the directly intervening habitat comprised 1 of 3 treatments: forested riparian corridor, linear living fencerow, or open pasture. Antshrikes returned faster and with greater success in riparian corridors relative to pasture treatments. This species also traveled more directly in riparian corridor treatments, detoured to use forested routes in the other 2 treatments, and did not use fencerows even when they led directly to their home territories. By contrast, wrens were more likely to use fencerows when returning, and return time and success were equivalent among the 3 treatments. Both species crossed fewer gaps in tree cover during riparian corridor treatments than in fencerow or pasture treatments. We conclude that antshrikes, which may be representative of other forest specialists, use forested corridors for movement in this landscape and that fencerows are avoided as movement conduits.

Structure and activity of full-length formin mDia1

PubMed Central

Maiti, Sankar; Michelot, Alphee; Gould, Christopher; Blanchoin, Laurent; Sokolova, Olga; Goode, Bruce L.

2012-01-01

Formins are a conserved family of actin assembly-promoting factors with essential and diverse biological roles. Most of our biochemical understanding of formin effects on actin dynamics is derived from studies using formin fragments. In addition, all structural information on formins has been limited to fragments. This has left open key questions about the structure, activity and regulation of intact formin proteins. Here, we isolated full-length mouse mDia1 (mDia1-FL) and found that it forms tightly autoinhibited dimers that can only be partially activated by RhoA. We solved the structure of autoinhibited mDia1-FL using electron microscopy and single particle analysis. Docking of crystal structures into the 3D reconstruction revealed that the fork-shaped N-terminal DID-CC region hangs over the ring-shaped FH2 domain, suggesting that autoinhibition results from steric obstruction of actin binding. Deletion of the C-terminal DAD domain extended mDia1 structure and activated it for actin assembly. Using TIRF microscopy, we observed that RhoA-activated mDia1-FL persistently accelerated filament elongation in the presence of profilin similar to mDia1 FH1-FH2 fragment. These observations validate the known activities of FH1-FH2 fragments as reflecting those of the intact molecule. Our results further suggest that mDia1-FL does not readily snap back into the autoinhibited conformation and dissociate from growing filament ends, and thus additional factors may be required to displace formins and restrict filament length. PMID:22605659

Modeling extracellular matrix degradation balance with proteinase/transglutaminase cycle.

PubMed

Larreta-Garde, Veronique; Berry, Hugues

2002-07-07

Extracellular matrix mass balance is implied in many physiological and pathological events, such as metastasis dissemination. Widely studied, its destructive part is mainly catalysed by extracellular proteinases. Conversely, the properties of the constructive part are less obvious, cellular neo-synthesis being usually considered as its only element. In this paper, we introduce the action of transglutaminase in a mathematical model for extracellular matrix remodeling. This extracellular enzyme, catalysing intermolecular protein cross-linking, is considered here as a reverse proteinase as far as the extracellular matrix physical state is concerned. The model is based on a proteinase/transglutaminase cycle interconverting insoluble matrix and soluble proteolysis fragments, with regulation of cellular proteinase expression by the fragments. Under "closed" (batch) conditions, i.e. neglecting matrix influx and fragment efflux from the system, the model is bistable, with reversible hysteresis. Extracellular matrix proteins concentration abruptly switches from low to high levels when transglutaminase activity exceeds a threshold value. Proteinase concentration usually follows the reverse complementary kinetics, but can become apparently uncoupled from extracellular matrix concentration for some parameter values. When matrix production by the cells and fragment degradation are taken into account, the dynamics change to sustained oscillations because of the emergence of a stable limit cycle. Transitions out of and into oscillation areas are controlled by the model parameters. Biological interpretation indicates that these oscillations could represent the normal homeostatic situation, whereas the other exhibited dynamics can be related to pathologies such as tumor invasion or fibrosis. These results allow to discuss the insights that the model could contribute to the comprehension of these complex biological events.

Diet and feeding ecology of the western hoolock gibbon (Hoolock hoolock) in a tropical forest fragment of Northeast India.

PubMed

Borah, Mrigakhi; Devi, Ashalata; Kumar, Awadhesh

2018-01-01

Forest fragmentation alters plant species diversity and composition, and causes diverse affects on the feeding behavior of wild primates. We investigated the feeding behavior and diet of two groups of western hoolock gibbon (Hoolock hoolock) inhabiting a small isolated forest patch (21 km 2 ) in Hollongapar Gibbon Wildlife Sanctuary, Assam, Northeast India, over a year using focal animal sampling. H. hoolock adults spent, on average, 35.2% of their total annual activity budget on feeding, and fed on young leaves, mature leaves, flowers, fruits, petioles, buds and also on animal matter. There was marked seasonal variation in the proportions of the dietary items consumed. Fruits accounted for an average of 51% (range 34-71% per month) of feeding time over the year. This highly frugivorous diet may limit the ability of the species to survive in small and disturbed forest fragments. A total of 54 plant species (32 families) were consumed by the focal groups during the study period, but there were variations between months in the selection of these plant species. Non-tree species such as lianas were among the most highly selected species in the diet. Moraceae, comprising ten species, was the most dominant family among the food plants, accounting for 36% of annual feeding time. The present study presents quantitative and qualitative data on dietary composition, preference and selection of food plants of H. hoolock in a fragmented habitat, which can contribute to the restoration and manipulation of degraded habitats of H. hoolock.

Introgression of wheat DNA markers from A, B and D genomes in early generation progeny of Aegilops cylindrica Host x Triticum aestivum L. hybrids.

PubMed

Schoenenberger, N; Felber, F; Savova-Bianchi, D; Guadagnuolo, R

2005-11-01

Introgression from allohexaploid wheat (Triticum aestivum L., AABBDD) to allotetraploid jointed goatgrass (Aegilops cylindrica Host, CCDD) can take place in areas where the two species grow in sympatry and hybridize. Wheat and Ae. cylindrica share the D genome, issued from the common diploid ancestor Aegilops tauschii Coss. It has been proposed that the A and B genome of bread wheat are secure places to insert transgenes to avoid their introgression into Ae. cylindrica because during meiosis in pentaploid hybrids, A and B genome chromosomes form univalents and tend to be eliminated whereas recombination takes place only in D genome chromosomes. Wheat random amplified polymorphic DNA (RAPD) fragments, detected in intergeneric hybrids and introgressed to the first backcross generation with Ae. cylindrica as the recurrent parent and having a euploid Ae. cylindrica chromosome number or one supernumerary chromosome, were assigned to wheat chromosomes using Chinese Spring nulli-tetrasomic wheat lines. Introgressed fragments were not limited to the D genome of wheat, but specific fragments of A and B genomes were also present in the BC1. Their presence indicates that DNA from any of the wheat genomes can introgress into Ae. cylindrica. Successfully located RAPD fragments were then converted into highly specific and easy-to-use sequence characterised amplified regions (SCARs) through sequencing and primer design. Subsequently these markers were used to characterise introgression of wheat DNA into a BC1S1 family. Implications for risk assessment of genetically modified wheat are discussed.

MALDI-MS/MS with Traveling Wave Ion Mobility for the Structural Analysis of N-Linked Glycans

NASA Astrophysics Data System (ADS)

Harvey, David J.; Scarff, Charlotte A.; Crispin, Max; Scanlan, Christopher N.; Bonomelli, Camille; Scrivens, James H.

2012-11-01

The preference for singly charged ion formation by MALDI makes it a better choice than electrospray ionization for profiling mixtures of N-glycans. For structural analysis, fragmentation of negative ions often yields more informative spectra than fragmentation of positive ones but such ions are more difficult to produce from neutral glycans under MALDI conditions. This work investigates conditions for the formation of both positive and negative ions by MALDI from N-linked glycans released from glycoproteins and their subsequent MS/MS and ion mobility behaviour. 2,4,6-Trihydroxyacetophenone (THAP) doped with ammonium nitrate was found to give optimal ion yields in negative ion mode. Ammonium chloride or phosphate also yielded prominent adducts but anionic carbohydrates such as sulfated N-glycans tended to ionize preferentially. Carbohydrates adducted with all three adducts (phosphate, chloride, and nitrate) produced good negative ion CID spectra but those adducted with iodide and sulfate did not yield fragment ions although they gave stronger signals. Fragmentation paralleled that seen following electrospray ionization providing superior spectra than could be obtained by PSD on MALDI-TOF instruments or with ion traps. In addition, ion mobility drift times of the adducted glycans and the ability of this technique to separate isomers also mirrored those obtained following ESI sample introduction. Ion mobility also allowed profiles to be obtained from samples whose MALDI spectra showed no evidence of such ions allowing the technique to be used in conditions where sample amounts were limiting. The method was applied to N-glycans released from the recombinant human immunodeficiency virus glycoprotein, gp120.

Black-and-white snub-nosed monkey (Rhinopithecus bieti) feeding behavior in a degraded forest fragment: clues to a stressed population.

PubMed

Huang, Zhi-Pang; Scott, Matthew B; Li, Yan-Peng; Ren, Guo-Peng; Xiang, Zuo-Fu; Cui, Liang-Wei; Xiao, Wen

2017-10-01

Rapid global deforestation has forced many of the world's primates to live in fragmented habitats, making the understanding of their behavioral responses to degraded and fragmented habitats a key challenge for their future protection and management. The black-and-white snub-nosed monkey (Rhinopithecus bieti) is an endangered species endemic to southwest China. The forest habitat ranges from near-continuous to fragmented. In this study, we investigated the activity budget and diet of a R. bieti population that live in an isolated and degraded habitat patch at Mt. Lasha in Yunnan Province, near the current southern limit of the species. We used our data along with data from six other sites in more-continuous habitats across its range to model factors that predict stress, including feeding effort and time feeding on lichens against potential predictive parameters. Models showed feeding effort across all sites increased with increasing altitude and latitude, and with decreasing food species diversity. There was also a strong positive relationship between feeding effort and time feeding lichens. The Mt. Lasha R. bieti population exploited a total of 36 food species, spending 80.2% of feeding time feeding on lichens, Bryoria spp. and Usnea longissima. These figures are more comparable to those living in the north than those living in the mid- and southern part of the species' range. Given the models for feeding effort and time feeding on lichens, the unexpectedly high time spend feeding on lichens and feeding effort relative to latitude and elevation are suggestive of a stressed population at Mt. Lasha.

Synchronous detection of ebolavirus conserved RNA sequences and ebolavirus-encoded miRNA-like fragment based on a zwitterionic copper (II) metal-organic framework.

PubMed

Qiu, Gui-Hua; Weng, Zi-Hua; Hu, Pei-Pei; Duan, Wen-Jun; Xie, Bao-Ping; Sun, Bin; Tang, Xiao-Yan; Chen, Jin-Xiang

2018-04-01

From a three-dimensional (3D) metal-organic framework (MOF) of {[Cu(Cmdcp)(phen)(H 2 O)] 2 ·9H 2 O} n (1, H 3 CmdcpBr = N-carboxymethyl-(3,5-dicarboxyl)pyridinium bromide, phen = phenanthroline), a sensitive and selective fluorescence sensor has been developed for the simultaneous detection of ebolavirus conserved RNA sequences and ebolavirus-encoded microRNA-like (miRNA-like) fragment. The results from molecular dynamics simulation confirmed that MOF 1 absorbs carboxyfluorescein (FAM)-tagged and 5(6)-carboxyrhodamine, triethylammonium salt (ROX)-tagged probe ss-DNA (probe DNA, P-DNA) by π … π stacking and hydrogen bonding, as well as additional electrostatic interactions to form a sensing platform of P-DNAs@1 with quenched FAM and ROX fluorescence. In the presence of targeted ebolavirus conserved RNA sequences or ebolavirus-encoded miRNA-like fragment, the fluorophore-labeled P-DNA hybridizes with the analyte to give a P-DNA@RNA duplex and released from MOF 1, triggering a fluorescence recovery. Simultaneous detection of two target RNAs has also been realized by single and synchronous fluorescence analysis. The formed sensing platform shows high sensitivity for ebolavirus conserved RNA sequences and ebolavirus-encoded miRNA-like fragment with detection limits at the picomolar level and high selectivity without cross-reaction between the two probes. MOF 1 thus shows the potential as an effective fluorescent sensing platform for the synchronous detection of two ebolavirus-related sequences, and offer improved diagnostic accuracy of Ebola virus disease. Copyright © 2017 Elsevier B.V. All rights reserved.

Functional traits variation explains the distribution of Aextoxicon punctatum (Aextoxicaceae) in pronounced moisture gradients within fog-dependent forest fragments.

PubMed

Salgado-Negret, Beatriz; Canessa, Rafaella; Valladares, Fernando; Armesto, Juan J; Pérez, Fernanda

2015-01-01

Climate change and fragmentation are major threats to world forests. Understanding how functional traits related to drought tolerance change across small-scale, pronounced moisture gradients in fragmented forests is important to predict species' responses to these threats. In the case of Aextoxicon punctatum, a dominant canopy tree in fog-dependent rain forest patches in semiarid Chile, we explored how the magnitude, variability and correlation patterns of leaf and xylem vessel traits and hydraulic conductivity varied across soil moisture (SM) gradients established within and among forest patches of different size, which are associated with differences in tree establishment and mortality patterns. Leaf traits varied across soil-moisture gradients produced by fog interception. Trees growing at drier leeward edges showed higher leaf mass per area, trichome and stomatal density than trees from the wetter core and windward zones. In contrast, xylem vessel traits (vessels diameter and density) did not vary producing loss of hydraulic conductivity at drier leeward edges. We also detected higher levels of phenotypic integration and variability at leeward edges. The ability of A. punctatum to modify leaf traits in response to differences in SM availability established over short distances (<500 m) facilitates its persistence in contrasting microhabitats within forest patches. However, xylem anatomy showed limited plasticity, which increases cavitation risk at leeward edges. Greater patch fragmentation, together with fluctuations in irradiance and SM in small patches, could result in higher risk of drought-related tree mortality, with profound impacts on hydrological balances at the ecosystem scale.

k(+)-buffer: An Efficient, Memory-Friendly and Dynamic k-buffer Framework.

PubMed

Vasilakis, Andreas-Alexandros; Papaioannou, Georgios; Fudos, Ioannis

2015-06-01

Depth-sorted fragment determination is fundamental for a host of image-based techniques which simulates complex rendering effects. It is also a challenging task in terms of time and space required when rasterizing scenes with high depth complexity. When low graphics memory requirements are of utmost importance, k-buffer can objectively be considered as the most preferred framework which advantageously ensures the correct depth order on a subset of all generated fragments. Although various alternatives have been introduced to partially or completely alleviate the noticeable quality artifacts produced by the initial k-buffer algorithm in the expense of memory increase or performance downgrade, appropriate tools to automatically and dynamically compute the most suitable value of k are still missing. To this end, we introduce k(+)-buffer, a fast framework that accurately simulates the behavior of k-buffer in a single rendering pass. Two memory-bounded data structures: (i) the max-array and (ii) the max-heap are developed on the GPU to concurrently maintain the k-foremost fragments per pixel by exploring pixel synchronization and fragment culling. Memory-friendly strategies are further introduced to dynamically (a) lessen the wasteful memory allocation of individual pixels with low depth complexity frequencies, (b) minimize the allocated size of k-buffer according to different application goals and hardware limitations via a straightforward depth histogram analysis and (c) manage local GPU cache with a fixed-memory depth-sorting mechanism. Finally, an extensive experimental evaluation is provided demonstrating the advantages of our work over all prior k-buffer variants in terms of memory usage, performance cost and image quality.

Functional traits variation explains the distribution of Aextoxicon punctatum (Aextoxicaceae) in pronounced moisture gradients within fog-dependent forest fragments

PubMed Central

Salgado-Negret, Beatriz; Canessa, Rafaella; Valladares, Fernando; Armesto, Juan J.; Pérez, Fernanda

2015-01-01

Climate change and fragmentation are major threats to world forests. Understanding how functional traits related to drought tolerance change across small-scale, pronounced moisture gradients in fragmented forests is important to predict species’ responses to these threats. In the case of Aextoxicon punctatum, a dominant canopy tree in fog-dependent rain forest patches in semiarid Chile, we explored how the magnitude, variability and correlation patterns of leaf and xylem vessel traits and hydraulic conductivity varied across soil moisture (SM) gradients established within and among forest patches of different size, which are associated with differences in tree establishment and mortality patterns. Leaf traits varied across soil-moisture gradients produced by fog interception. Trees growing at drier leeward edges showed higher leaf mass per area, trichome and stomatal density than trees from the wetter core and windward zones. In contrast, xylem vessel traits (vessels diameter and density) did not vary producing loss of hydraulic conductivity at drier leeward edges. We also detected higher levels of phenotypic integration and variability at leeward edges. The ability of A. punctatum to modify leaf traits in response to differences in SM availability established over short distances (<500 m) facilitates its persistence in contrasting microhabitats within forest patches. However, xylem anatomy showed limited plasticity, which increases cavitation risk at leeward edges. Greater patch fragmentation, together with fluctuations in irradiance and SM in small patches, could result in higher risk of drought-related tree mortality, with profound impacts on hydrological balances at the ecosystem scale. PMID:26257746

Collaborative study on the effect of grinding on the detection of bones from processed animal proteins in feed by light microscopy.

PubMed

Veys, Pascal; Planchon, Viviane; Colbert, Ruairi; Cruz, Clara; Frick, Geneviève; Ioannou, Ioannis; Marchis, Daniela; Nordkvist, Erik; Paradies-Severin, Inge; Pohto, Arja; Weiss, Roland; Baeten, Vincent; Berben, Gilbert

2017-08-01

Bone fragments are essential structures for the detection of processed animal proteins (PAPs) in feed by light microscopy for official controls according to Annex VI of European Union Regulation EC/152/2009. The preparation of samples submitted for analysis requires a grinding step to make them suitable for microscopic slide preparation and observation. However, there are no technical guidelines set down for this step despite the fact that it can lead to an increase in bone numbers due to fragmentation. This was demonstrated by an in-house study carried out by the Irish National Reference Laboratory (NRL) for animal protein detection. The present collaborative study investigated the possible effects of three different grinding conditions on the final result for a feed adulterated with 0.05 and 0.01% (w/w) of PAP. The microscopic analysis either combined or not with an Alizarin Red staining was carried out by 10 different laboratories. The results demonstrated that although a large variation in the numbers of bone fragments was noted, five of the six different grinding/staining combinations applied at two levels of PAP adulteration did not significantly (at p = 0.05) differ from one another. The only exception occurred when grinding the feed containing 0.05% of PAP with a rotor mill equipped with a 0.5-mm sieve and combined with a staining which resulted in a greater number of bone fragments by forced fragmentation. Overall, the impact of the grinding/staining combinations on the final results was shown to be negligible when considering the regulatory limit of detection (LOD) requirement for the method and the current rules of implementation of the light microscopic method. From a total of 180 analyses carried out on the feed matrix containing 0.05% of PAP no false-negative result was observed, and at a level of 0.01% PAP only 10 false-negative results occurred.

Colliding fragment islands transport independent lineages of endemic rock-crawlers (Grylloblattodea: Grylloblattidae) in the Japanese archipelago.

PubMed

Schoville, Sean D; Uchifune, Toshiki; Machida, Ryuichiro

2013-03-01

Fragment islands, viewed from the paradigm of island biogeographic theory, depend on continual immigration from continental sources to maintain levels of species diversity, or otherwise undergo a period of relaxation where species diversity declines to a lower equilibrium. Japan is a recently derived fragment island with a rich endemic flora and fauna. These endemic species have been described as paleoendemics, and conversely as recently derived Pleistocene colonists. Geological events in the Miocene period, notably the fragmentation and collision of islands, and the subsequent uplift of mountains in central Japan, provided opportunities for genetic isolation. More recently, cyclical climatic change during the Pliocene and Pleistocene periods led to intermittent land bridge connections to continental Asia. Here we investigate the pattern and timing of diversification in a diverse endemic lineage in order to test whether ongoing migration has sustained species diversity, whether there is evidence of relaxation, and how geological and climatic events are associated with lineage diversification. Using multi-locus genetic data, we test these hypotheses in a poorly dispersing, cold-adapted terrestrial insect lineage (Grylloblattodea: Grylloblattidae) sampled from Japan, Korea, and Russia. In phylogenetic analyses of concatenated data and a species tree approach, we find evidence of three deeply divergent lineages of rock-crawlers in Japan consistent with the pattern of island fragmentation from continental Asia. Tests of lineage diversification rates suggest that relaxation has not occurred and instead endemism has increased in the Japanese Grylloblattidae following mountain-building events in the Miocene. Although the importance of climate change in generating species diversity is a commonly held paradigm in Japanese biogeography, our analyses, including analyses of demographic change and phylogeographic range shifts in putative species, suggests that Pleistocene climatic change has had a limited effect on the diversification of rock-crawlers. Copyright © 2012 Elsevier Inc. All rights reserved.

Amidase Activity of AmiC Controls Cell Separation and Stem Peptide Release and Is Enhanced by NlpD in Neisseria gonorrhoeae.

PubMed

Lenz, Jonathan D; Stohl, Elizabeth A; Robertson, Rosanna M; Hackett, Kathleen T; Fisher, Kathryn; Xiong, Kalia; Lee, Mijoon; Hesek, Dusan; Mobashery, Shahriar; Seifert, H Steven; Davies, Christopher; Dillard, Joseph P

2016-05-13

The human-restricted pathogen Neisseria gonorrhoeae encodes a single N-acetylmuramyl-l-alanine amidase involved in cell separation (AmiC), as compared with three largely redundant cell separation amidases found in Escherichia coli (AmiA, AmiB, and AmiC). Deletion of amiC from N. gonorrhoeae results in severely impaired cell separation and altered peptidoglycan (PG) fragment release, but little else is known about how AmiC functions in gonococci. Here, we demonstrated that gonococcal AmiC can act on macromolecular PG to liberate cross-linked and non-cross-linked peptides indicative of amidase activity, and we provided the first evidence that a cell separation amidase can utilize a small synthetic PG fragment as substrate (GlcNAc-MurNAc(pentapeptide)-GlcNAc-MurNAc(pentapeptide)). An investigation of two residues in the active site of AmiC revealed that Glu-229 is critical for both normal cell separation and the release of PG fragments by gonococci during growth. In contrast, Gln-316 has an autoinhibitory role, and its mutation to lysine resulted in an AmiC with increased enzymatic activity on macromolecular PG and on the synthetic PG derivative. Curiously, the same Q316K mutation that increased AmiC activity also resulted in cell separation and PG fragment release defects, indicating that activation state is not the only factor determining normal AmiC activity. In addition to displaying high basal activity on PG, gonococcal AmiC can utilize metal ions other than the zinc cofactor typically used by cell separation amidases, potentially protecting its ability to function in zinc-limiting environments. Thus gonococcal AmiC has distinct differences from related enzymes, and these studies revealed parameters for how AmiC functions in cell separation and PG fragment release. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

The rate of polymerase release upon filling the gap between Okazaki fragments is inadequate to support cycling during lagging strand synthesis.

PubMed

Dohrmann, Paul R; Manhart, Carol M; Downey, Christopher D; McHenry, Charles S

2011-11-18

Upon completion of synthesis of an Okazaki fragment, the lagging strand replicase must recycle to the next primer at the replication fork in under 0.1 s to sustain the physiological rate of DNA synthesis. We tested the collision model that posits that cycling is triggered by the polymerase encountering the 5'-end of the preceding Okazaki fragment. Probing with surface plasmon resonance, DNA polymerase III holoenzyme initiation complexes were formed on an immobilized gapped template. Initiation complexes exhibit a half-life of dissociation of approximately 15 min. Reduction in gap size to 1 nt increased the rate of dissociation 2.5-fold, and complete filling of the gap increased the off-rate an additional 3-fold (t(1/2)~2 min). An exogenous primed template and ATP accelerated dissociation an additional 4-fold in a reaction that required complete filling of the gap. Neither a 5'-triphosphate nor a 5'-RNA terminated oligonucleotide downstream of the polymerase accelerated dissociation further. Thus, the rate of polymerase release upon gap completion and collision with a downstream Okazaki fragment is 1000-fold too slow to support an adequate rate of cycling and likely provides a backup mechanism to enable polymerase release when the other cycling signals are absent. Kinetic measurements indicate that addition of the last nucleotide to fill the gap is not the rate-limiting step for polymerase release and cycling. Modest (approximately 7 nt) strand displacement is observed after the gap between model Okazaki fragments is filled. To determine the identity of the protein that senses gap filling to modulate affinity of the replicase for the template, we performed photo-cross-linking experiments with highly reactive and non-chemoselective diazirines. Only the α subunit cross-linked, indicating that it serves as the sensor. Copyright © 2011 Elsevier Ltd. All rights reserved.

Fixation Options for the Volar Lunate Facet Fracture: Thinking Outside the Box.

PubMed

Harness, Neil G

2016-03-01

Background Fractures of the distal radius with small volar ulnar marginal fracture fragments are difficult to stabilize with standard volar locking plates. The purpose of this study is to describe alternative techniques available to stabilize these injuries. Materials and Methods Five patients were identified retrospectively with unstable volar lunate facet fracture fragments treated with supplemental fixation techniques. The demographic data, pre- and postoperative radiographic parameters, and early outcomes data were analyzed. The AO classification, preoperative and final postoperative ulnar variance, articular step-off, volar tilt, radial inclination, and teardrop angle were measured. The lunate subsidence and length of the volar cortex available for fixation were measured from the initial injury films. Description of Technique Lunate facet fixation was based on the morphology of the fragment, and stabilization was achieved with headless compression screws in three patients, a tension band wire construct in one, and two cortical screws in another. Results Five patients with a mean age of 58 years (range: 41-82) were included. There were two AO C3.2 and three B3.3 fractures. Preoperative radiographic measurements including radial inclination, tilt, and ulnar variance all improved after surgery and were maintained within normal limits at 3-month follow-up. There was no change in the teardrop angle at final follow-up (70-64 degrees; p = 0.14). None of the patients had loss of fixation or volar carpal subluxation. The mean visual analog scale pain score at 3 months was 1 (range: 0-2). Conclusions The morphology of volar lunate facet fracture fragments is variable, and fixation must be customized to the particular pattern. Small fragments may preclude the use of plates and screws for fixation. These fractures can be managed successfully with tension band wire constructs and headless screws. These low-profile implants may decrease the risk of tendon irritation that might accompany distally placed plates.

Fixation Options for the Volar Lunate Facet Fracture: Thinking Outside the Box

PubMed Central

Harness, Neil G.

2016-01-01

Background Fractures of the distal radius with small volar ulnar marginal fracture fragments are difficult to stabilize with standard volar locking plates. The purpose of this study is to describe alternative techniques available to stabilize these injuries. Materials and Methods Five patients were identified retrospectively with unstable volar lunate facet fracture fragments treated with supplemental fixation techniques. The demographic data, pre- and postoperative radiographic parameters, and early outcomes data were analyzed. The AO classification, preoperative and final postoperative ulnar variance, articular step-off, volar tilt, radial inclination, and teardrop angle were measured. The lunate subsidence and length of the volar cortex available for fixation were measured from the initial injury films. Description of Technique Lunate facet fixation was based on the morphology of the fragment, and stabilization was achieved with headless compression screws in three patients, a tension band wire construct in one, and two cortical screws in another. Results Five patients with a mean age of 58 years (range: 41–82) were included. There were two AO C3.2 and three B3.3 fractures. Preoperative radiographic measurements including radial inclination, tilt, and ulnar variance all improved after surgery and were maintained within normal limits at 3-month follow-up. There was no change in the teardrop angle at final follow-up (70–64 degrees; p = 0.14). None of the patients had loss of fixation or volar carpal subluxation. The mean visual analog scale pain score at 3 months was 1 (range: 0–2). Conclusions The morphology of volar lunate facet fracture fragments is variable, and fixation must be customized to the particular pattern. Small fragments may preclude the use of plates and screws for fixation. These fractures can be managed successfully with tension band wire constructs and headless screws. These low-profile implants may decrease the risk of tendon irritation that might accompany distally placed plates. PMID:26855830

Randomized DNA libraries construction tool: a new 3-bp 'frequent cutter' TthHB27I/sinefungin endonuclease with chemically-induced specificity.

PubMed

Krefft, Daria; Papkov, Aliaksei; Prusinowski, Maciej; Zylicz-Stachula, Agnieszka; Skowron, Piotr M

2018-05-11

Acoustic or hydrodynamic shearing, sonication and enzymatic digestion are used to fragment DNA. However, these methods have several disadvantages, such as DNA damage, difficulties in fragmentation control, irreproducibility and under-representation of some DNA segments. The DNA fragmentation tool would be a gentle enzymatic method, offering cleavage frequency high enough to eliminate DNA fragments distribution bias and allow for easy control of partial digests. Only three such frequently cleaving natural restriction endonucleases (REases) were discovered: CviJI, SetI and FaiI. Therefore, we have previously developed two artificial enzymatic specificities, cleaving DNA approximately every ~ 3-bp: TspGWI/sinefungin (SIN) and TaqII/SIN. In this paper we present the third developed specificity: TthHB27I/SIN(SAM) - a new genomic tool, based on Type IIS/IIC/IIG Thermus-family REases-methyltransferases (MTases). In the presence of dimethyl sulfoxide (DMSO) and S-adenosyl-L-methionine (SAM) or its analogue SIN, the 6-bp cognate TthHB27I recognition sequence 5'-CAARCA-3' is converted into a combined 3.2-3.0-bp 'site' or its statistical equivalent, while a cleavage distance of 11/9 nt is retained. Protocols for various modes of limited DNA digestions were developed. In the presence of DMSO and SAM or SIN, TthHB27I is transformed from rare 6-bp cutter to a very frequent one, approximately 3-bp. Thus, TthHB27I/SIN(SAM) comprises a new tool in the very low-represented segment of such prototype REases specificities. Moreover, this modified TthHB27I enzyme is uniquely suited for controlled DNA fragmentation, due to partial DNA cleavage, which is an inherent feature of the Thermus-family enzymes. Such tool can be used for quasi-random libraries generation as well as for other DNA manipulations, requiring high frequency cleavage and uniform distribution of cuts along DNA.

Genetic analysis of 430 Chinese Cynodon dactylon accessions using sequence-related amplified polymorphism markers.

PubMed

Huang, Chunqiong; Liu, Guodao; Bai, Changjun; Wang, Wenqiang

2014-10-21

Although Cynodon dactylon (C. dactylon) is widely distributed in China, information on its genetic diversity within the germplasm pool is limited. The objective of this study was to reveal the genetic variation and relationships of 430 C. dactylon accessions collected from 22 Chinese provinces using sequence-related amplified polymorphism (SRAP) markers. Fifteen primer pairs were used to amplify specific C. dactylon genomic sequences. A total of 481 SRAP fragments were generated, with fragment sizes ranging from 260-1800 base pairs (bp). Genetic similarity coefficients (GSC) among the 430 accessions averaged 0.72 and ranged from 0.53-0.96. Cluster analysis conducted by two methods, namely the unweighted pair-group method with arithmetic averages (UPGMA) and principle coordinate analysis (PCoA), separated the accessions into eight distinct groups. Our findings verify that Chinese C. dactylon germplasms have rich genetic diversity, which is an excellent basis for C. dactylon breeding for new cultivars.

Palaeosymbiosis Revealed by Genomic Fossils of Wolbachia in a Strongyloidean Nematode

PubMed Central

Koutsovoulos, Georgios; Makepeace, Benjamin; Tanya, Vincent N.; Blaxter, Mark

2014-01-01

Wolbachia are common endosymbionts of terrestrial arthropods, and are also found in nematodes: the animal-parasitic filaria, and the plant-parasite Radopholus similis. Lateral transfer of Wolbachia DNA to the host genome is common. We generated a draft genome sequence for the strongyloidean nematode parasite Dictyocaulus viviparus, the cattle lungworm. In the assembly, we identified nearly 1 Mb of sequence with similarity to Wolbachia. The fragments were unlikely to derive from a live Wolbachia infection: most were short, and the genes were disabled through inactivating mutations. Many fragments were co-assembled with definitively nematode-derived sequence. We found limited evidence of expression of the Wolbachia-derived genes. The D. viviparus Wolbachia genes were most similar to filarial strains and strains from the host-promiscuous clade F. We conclude that D. viviparus was infected by Wolbachia in the past, and that clade F-like symbionts may have been the source of filarial Wolbachia infections. PMID:24901418

Seismic evidence for rotating mantle flow around subducting slab edge associated with oceanic microplate capture

NASA Astrophysics Data System (ADS)

Mosher, Stephen G.; Audet, Pascal; L'Heureux, Ivan

2014-07-01

Tectonic plate reorganization at a subduction zone edge is a fundamental process that controls oceanic plate fragmentation and capture. However, the various factors responsible for these processes remain elusive. We characterize seismic anisotropy of the upper mantle in the Explorer region at the northern limit of the Cascadia subduction zone from teleseismic shear wave splitting measurements. Our results show that the mantle flow field beneath the Explorer slab is rotating anticlockwise from the convergence-parallel motion between the Juan de Fuca and the North America plates, re-aligning itself with the transcurrent motion between the Pacific and North America plates. We propose that oceanic microplate fragmentation is driven by slab stretching, thus reorganizing the mantle flow around the slab edge and further contributing to slab weakening and increase in buoyancy, eventually leading to cessation of subduction and microplate capture.

A Review on Assessment and Treatment for Depression in Malaysia

PubMed Central

Mukhtar, Firdaus; Oei, Tian P. S.

2011-01-01

This paper aimed to review the literature on depression that focused on its assessment and treatment in Malaysia. PsycINFO, Medline, local journals were searched, and 18 published articles were included in this paper. Results indicate that research on depression in Malaysia, particularly validation studies and psychotherapy research, was weak and fragmented, with minimal empirical evidence available. Pharmacotherapy still dominated the treatment for depression, and, in terms of psychotherapy, Cognitive Behavioural Therapy (CBT) was recently practiced, but only a few studies have reported on the treatment efficacy of CBT. Major limitations of studies were noted, and, consequently, the problems that are associated with the implementation and future direction of clinical and research on depression in Malaysia were discussed. In short, the contribution of empirical research on the assessment and treatment for depression remained inconsistent and fragmented and urgently in need of further empirical investigation. PMID:21804939

NIOSH testimony to DOL on the Occupational Safety and Health Administration's notice of proposed rulemaking on occupational exposure to asbestos, tremolite, anthophyllite, and actinolite by R. A. Lemen, May 9, 1990

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1990-05-09

The testimony summarized information pertinent to the proposed rulemaking to remove nonasbestiform tremolite, anthophyllite, and actinolite from the asbestos standard. NIOSH concludes that on the basis of current data that cleavage fragments of the appropriate aspect ratio and length from the nonasbestiform minerals should be considered as hazardous as fibers from the asbestiform minerals. No scientifically valid health evidence was found for removing from the asbestos standard cleavage fragments that become airborne when nonasbestiform tremolite, anthophyllite and actinolite are mined, milled and used, and that meet the microscopic definition of a fiber. The risk of cancer from such exposures warrantsmore » limiting exposures to these minerals to the lowest feasible concentration. A glossary of terms is provided in an appendix.« less

Reactions of methyl groups on a non-reducible metal oxide: The reaction of iodomethane on stoichiometric α-Cr 2O 3(0001)

DOE PAGES

Dong, Yujung; Brooks, John D.; Chen, Tsung-Liang; ...

2015-06-10

The reaction of iodomethane on the nearly stoichiometric α-Cr 2O 3(0001) surface produces gas phase ethylene, methane, and surface iodine adatoms. The reaction is first initiated by the dissociation of iodomethane into surface methyl fragments, -CH 3, and iodine adatoms. Methyl fragments bound at surface Cr cation sites undergo a rate-limiting dehydrogenation reaction to methylene, =CH 2. The methylene intermediates formed from methyl dehydrogenation can then undergo coupling reactions to produce ethylene via two principle reaction pathways: (1) direct coupling of methylene and (2) methylene insertion into the methyl surface bond to form surface ethyl groups which undergo β-H eliminationmore » to produce ethylene. The liberated hydrogen also combines with methyl groups to form methane. Iodine adatoms from the dissociation of iodomethane deactivate the surface by simple site blocking of the surface Cr 3+ cations.« less

The fundholding fandango.

PubMed Central

Bowie, C; Harris, T

1994-01-01

General practitioner fundholding allows flexible use of resources at the coal-face, provides incentives to alter practice such as prescribing within cash limits and forces hospitals to be more responsive to general practitioner demands. However, the additional administrative costs both in time and money, the fragmentation of purchasing power compounded by a lack of expertise and experience in contracting, and the poor information and financial systems which exist in the National Health Service are severe constraints. A suggested way forward is to delegate responsibility for running the scheme, including the contracting and billing, to district health authorities offering more flexible budgets to all practices and extending the scheme as local information systems allow. This will reduce fragmentation of purchasing power and administrative costs and re-establish local accountability. It will also give the general practitioner more time to see and treat patients, who will see the system as being fairer. PMID:8312038

The Myxococcus xanthus Spore Cuticula Protein C Is a Fragment of FibA, an Extracellular Metalloprotease Produced Exclusively in Aggregated Cells

PubMed Central

Lee, Bongsoo; Mann, Petra; Grover, Vidhi; Treuner-Lange, Anke; Kahnt, Jörg; Higgs, Penelope I.

2011-01-01

Myxococcus xanthus is a soil bacterium with a complex life cycle involving distinct cell fates, including production of environmentally resistant spores to withstand periods of nutrient limitation. Spores are surrounded by an apparently self-assembling cuticula containing at least Proteins S and C; the gene encoding Protein C is unknown. During analyses of cell heterogeneity in M. xanthus, we observed that Protein C accumulated exclusively in cells found in aggregates. Using mass spectrometry analysis of Protein C either isolated from spore cuticula or immunoprecipitated from aggregated cells, we demonstrate that Protein C is actually a proteolytic fragment of the previously identified but functionally elusive zinc metalloprotease, FibA. Subpopulation specific FibA accumulation is not due to transcriptional regulation suggesting post-transcriptional regulation mechanisms mediate its heterogeneous accumulation patterns. PMID:22174937

Mycobacterium avium restriction fragment length polymorphism-IS IS1245 and the simple double repetitive element polymerase chain reaction typing method to screen genetic diversity in Brazilian strains.

PubMed

Sequeira, Patrícia Carvalho de; Fonseca, Leila de Souza; Silva, Marlei Gomes da; Saad, Maria Helena Féres

2005-11-01

Simple double repetitive element polymerase chain reaction (MaDRE-PCR) and Pvu II-IS1245 restriction fragment length polymorphism (RFLP) typing methods were used to type 41 Mycobacterium avium isolates obtained from 14 AIDS inpatients and 10 environment and animals specimens identified among 53 mycobacteria isolated from 237 food, chicken, and pig. All environmental and animals strains showed orphan patterns by both methods. By MaDRE-PCR four patients, with multiple isolates, showed different patterns, suggesting polyclonal infection that was confirmed by RFLP in two of them. This first evaluation of MaDRE-PCR on Brazilian M. avium strains demonstrated that the method seems to be useful as simple and less expensive typing method for screening genetic diversity in M. avium strains on selected epidemiological studies, although with limitation on analysis identical patterns except for one band.

Elbow arthroscopy: valgus extension overload.

PubMed

Ahmad, Christopher S; Conway, John E

2011-01-01

Valgus torque combined with deceleration produces high compression and shear forces acting on the posteromedial olecranon and the posteromedial trochlea. This valgus extension overload process may cause posteromedial trochlea chondromalacia, chondral flap formation, osteochondrosis, subchondral erosion, a subchondral insufficiency fracture, and marginal exostosis formation. Olecranon pathologies include proximal stress reaction, a posteromedial tip stress fracture, a transverse proximal process stress fracture, exostosis formation, exostosis fragmentation, and intra-articular loose bodies. Symptoms include posteromedial elbow pain during the deceleration phase of the throwing motion. The extension impingement test reproduces posterior or posteromedial pain similar to that experienced while throwing. Special radiographic techniques and CT scans can show loose bodies and osteophyte fragmentation. Surgical treatment is indicated when symptoms persist despite nonsurgical management. Based on clinical and basic science research, all patients with valgus extension overload should be comprehensively evaluated for medial ulnar collateral ligament insufficiency. Surgical treatment is limited to the resection of osteophytes only; normal olecranon should not be resected.

Mass Spectra of Some Perfluoroalkyl and Perfluoroalkylether Substituted 1,2,4-Oxadiazoles

NASA Technical Reports Server (NTRS)

Paciorek, Kazimiera J. L.; Nakahara, James H.; Kratzer, Reinhold H.; Rosser, Robert W.

1977-01-01

Electron impact fragmentation patterns were obtained for 1,4-bis[(5-perfluoro-n-heptyl)-1,2,4-oxadiazolyl- benzene, its perfluoroalkylether substituted analogue, 3,5-bis(perfluoroalkyl)-, 3,5-bis(perfluoroalkylether)- and 3-perfluoroalkylether-5-perfluoroalkyl-1,2,4-oxadiazoles. In the compounds containing the phenylene group the molecular ion constituted the base peak; the main process was the breakdown of the oxadiazole ring with concurrent liberation of the perfluoroalkyl or perfluoroalkylether nitrile molecule; cleavage of the fluorinated chain ot to the oxadiazole ring was found to take place to a considerable degree. In the perfluorinated 1,2,4-oxadiazoles cleavage beta to the oxadiazole ring occurred preferentially; fragmentation of the ring itself took place to a limited degree only. The 3-perfluoroalkylether-5-perfluoroalkyl-1,2,4-oxadiazole appeared to undergo the primary beta-cleavage exclusively at the perfluoroalkylether sidechain.

Ultrastructure of the membrana limitans interna after dye-assisted membrane peeling.

PubMed

Brockmann, Tobias; Steger, Claudia; Westermann, Martin; Nietzsche, Sandor; Koenigsdoerffer, Ekkehart; Strobel, Juergen; Dawczynski, Jens

2011-01-01

The purpose of this study was to investigate the ultrastructure of the membrana limitans interna (internal limiting membrane, ILM) and to evaluate alterations to the retinal cell layers after membrane peeling with vital dyes. Twenty-five patients (25 eyes) who underwent macular hole surgery were included, whereby 12 indocyanine green (ICG)- and 13 brilliant blue G (BBG)-stained ILM were analyzed using light, transmission electron and scanning electron microscopy. Retinal cell fragments on the ILM were identified in both groups using immunohistochemistry. Comparing ICG- and BBG-stained membranes, larger cellular fragments were observed at a higher frequency in the BBG group. Thereby, the findings indicate that ICG permits an enhanced separation of the ILM from the underlying retina with less mechanical destruction. A possible explanation might be seen in the known photosensitivity of ICG, which induces a stiffening and shrinkage of the ILM but also generates retinal toxic metabolites. Copyright © 2011 S. Karger AG, Basel.

Retention of polyurethane foam fragments during VAC therapy: a complication to be considered.

PubMed

Dessy, Luca A; Serratore, Francesco; Corrias, Federico; Parisi, Paola; Mazzocchi, Marco; Carlesimo, Bruno

2015-04-01

Vacuum-assisted closure (VAC) therapy is a closed-loop, non-invasive active system, characterised by a controlled and localised negative pressure applied on porous polyurethane absorbent foams. It promotes healing of acute and chronic wounds. Therapeutic effects of VAC therapy have been proved and demonstrated; however, this method can have some disadvantages. Even if it is a quite versatile device, only qualified medical/paramedical personnel should use it in order to avoid possible complications that can occur after an improper application. In this report, 11 cases of foam-fragment retention within the wound are presented. This rare complication did not promote healing, but further hindered it. On the basis of our experience, it is mandatory to define the indications, benefits and limitations of VAC therapy. © 2013 The Authors. International Wound Journal © 2013 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

Local fragmentation of thin disks in Eddington-inspired gravity

NASA Astrophysics Data System (ADS)

Roshan, Mahmood; Kazemi, Ali; De Martino, Ivan

2018-06-01

We find the generalized version of the Toomre's criterion for the stability of a rotating thin disk in the context of Eddington inspired Born-Infeld (EiBI) gravity which possesses one free parameter χ. To do so we use the weak field limit of the theory and find the dispersion relation for the propagation of matter density waves on the surface of a self-gravitating and differentially rotating disk. Finally we find a new version of Toomre's stability criterion for thin disks. We show that EiBI gravity with negative χ destabilizes all the rotating thin disks. On the other hand EiBI with positive χ substantially can suppress the local fragmentation, and has stabilizing effects against axi-symmetric perturbations. More specifically, we show that only an annulus remains unstable on the surface of the disk. The width of the annulus directly depends on the magnitude of χ.

Mapping of the Available Chemical Space versus the Chemical Universe of Lead-Like Compounds.

PubMed

Lin, Arkadii; Horvath, Dragos; Afonina, Valentina; Marcou, Gilles; Reymond, Jean-Louis; Varnek, Alexandre

2018-03-20

This is, to our knowledge, the most comprehensive analysis to date based on generative topographic mapping (GTM) of fragment-like chemical space (40 million molecules with no more than 17 heavy atoms, both from the theoretically enumerated GDB-17 and real-world PubChem/ChEMBL databases). The challenge was to prove that a robust map of fragment-like chemical space can actually be built, in spite of a limited (≪10 5 ) maximal number of compounds ("frame set") usable for fitting the GTM manifold. An evolutionary map building strategy has been updated with a "coverage check" step, which discards manifolds failing to accommodate compounds outside the frame set. The evolved map has a good propensity to separate actives from inactives for more than 20 external structure-activity sets. It was proven to properly accommodate the entire collection of 40 m compounds. Next, it served as a library comparison tool to highlight biases of real-world molecules (PubChem and ChEMBL) versus the universe of all possible species represented by FDB-17, a fragment-like subset of GDB-17 containing 10 million molecules. Specific patterns, proper to some libraries and absent from others (diversity holes), were highlighted. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Does fragmentation of Urtica habitats affect phytophagous and predatory insects differentially?

PubMed

Zabel, Jörg; Tscharntke, Teja

1998-09-01

Effects of habitat fragmentation on the insect community of stinging nettle (Urtica dioica L.) were studied, using 32 natural nettle patches of different area and degree of isolation in an agricultural landscape. Habitat fragmentation reduced the species richness of Heteroptera, Auchenorrhyncha, and Coleoptera, and the abundance of populations. Habitat isolation and area reduction did not affect all insect species equally. Monophagous herbivores had a higher probability of absence from small patches than all (monophagous and polyphagous) herbivore species, and the percentage of monophagous herbivores increased with habitat area. Abundance and population variability of species were negatively correlated and could both be used as a predictor of the percentage of occupied habitats. Species richness of herbivores correlated (positively) with habitat area, while species richness of predators correlated (negatively) with habitat isolation. In logistic regressions, the probability of absence of monophagous herbivores from habitat patches could only be explained by habitat area (in 4 out of 10 species) and predator absence probability only by habitat isolation (in 3 out of 14 species). Presumably because of the instability of higher-trophic-level populations and dispersal limitation, predators were more affected by habitat isolation than herbivores, while they did not differ from herbivore populations with respect to abundance or variability. Thus increasing habitat connectivity in the agricultural landscape should primarily promote predator populations.

Rapid enzyme immunoassay for determination of toxigenicity among clinical isolates of corynebacteria.

PubMed

Engler, K H; Efstratiou, A

2000-04-01

A rapid enzyme immunoassay (EIA) was developed for the phenotypic detection of diphtheria toxin among clinical isolates of corynebacteria. The assay uses equine polyclonal antitoxin as the capture antibody and an alkaline phosphatase-labeled monoclonal antibody, specific for fragment A of the toxin molecule, as the detecting antibody. The assay is rapid, sensitive, and specific: a final result is available within 3 h of colony selection, and the limits of detection are 0.1 ng of pure diphtheria toxin/ml. Toxigenicity could be detected with isolates grown on a diverse range of culture media, including selective agars. Toxin detection using the EIA was compared to that with the Elek test and PCR detection of fragment A of the diphtheria toxin (tox) gene, using 245 isolates of corynebacteria. The results for the EIA were in complete concordance with those of the Elek test: 87 toxigenic and 158 nontoxigenic isolates. Ten of the phenotypically nontoxigenic strains were found to contain fragment A of the tox gene but did not express the toxin protein. These isolates were found to be nontoxigenic in the Vero cell tissue culture cytotoxicity assay and were therefore nontoxigenic for diagnostic purposes. The EIA is a simple rapid phenotypic test which provides a definitive result on toxigenicity within one working day.

Rapid Enzyme Immunoassay for Determination of Toxigenicity among Clinical Isolates of Corynebacteria

PubMed Central

Engler, Kathryn H.; Efstratiou, Androulla

2000-01-01

A rapid enzyme immunoassay (EIA) was developed for the phenotypic detection of diphtheria toxin among clinical isolates of corynebacteria. The assay uses equine polyclonal antitoxin as the capture antibody and an alkaline phosphatase-labeled monoclonal antibody, specific for fragment A of the toxin molecule, as the detecting antibody. The assay is rapid, sensitive, and specific: a final result is available within 3 h of colony selection, and the limits of detection are 0.1 ng of pure diphtheria toxin/ml. Toxigenicity could be detected with isolates grown on a diverse range of culture media, including selective agars. Toxin detection using the EIA was compared to that with the Elek test and PCR detection of fragment A of the diphtheria toxin (tox) gene, using 245 isolates of corynebacteria. The results for the EIA were in complete concordance with those of the Elek test: 87 toxigenic and 158 nontoxigenic isolates. Ten of the phenotypically nontoxigenic strains were found to contain fragment A of the tox gene but did not express the toxin protein. These isolates were found to be nontoxigenic in the Vero cell tissue culture cytotoxicity assay and were therefore nontoxigenic for diagnostic purposes. The EIA is a simple rapid phenotypic test which provides a definitive result on toxigenicity within one working day. PMID:10747112

Duplex microfluidic SERS detection of pathogen antigens with nanoyeast single-chain variable fragments.

PubMed

Wang, Yuling; Rauf, Sakandar; Grewal, Yadveer S; Spadafora, Lauren J; Shiddiky, Muhammad J A; Cangelosi, Gerard A; Schlücker, Sebastian; Trau, Matt

2014-10-07

Quantitative and accurate detection of multiple biomarkers would allow for the rapid diagnosis and treatment of diseases induced by pathogens. Monoclonal antibodies are standard affinity reagents applied for biomarkers detection; however, their production is expensive and labor-intensive. Herein, we report on newly developed nanoyeast single-chain variable fragments (NYscFv) as an attractive alternative to monoclonal antibodies, which offers the unique advantage of a cost-effective production, stability in solution, and target-specificity. By combination of surface-enhanced Raman scattering (SERS) microspectroscopy using glass-coated, highly purified SERS nanoparticle clusters as labels, with a microfluidic device comprising multiple channels, a robust platform for the sensitive duplex detection of pathogen antigens has been developed. Highly sensitive detection for individual Entamoeba histolytica antigen EHI_115350 (limit of detection = 1 pg/mL, corresponding to 58.8 fM) and EHI_182030 (10 pg/mL, corresponding 453 fM) with high specificity has been achieved, employing the newly developed corresponding NYscFv as probe in combination with SERS microspectroscopy at a single laser excitation wavelength. Our first report on SERS-based immunoassays using the novel NYscFv affinity reagent demonstrates the flexibility of NYscFv fragments as viable alternatives to monoclonal antibodies in a range of bioassay platforms and paves the way for further applications.

First assembly times and equilibration in stochastic coagulation-fragmentation

DOE Office of Scientific and Technical Information (OSTI.GOV)

D’Orsogna, Maria R.; Department of Mathematics, CSUN, Los Angeles, California 91330-8313; Lei, Qi

2015-07-07

We develop a fully stochastic theory for coagulation and fragmentation (CF) in a finite system with a maximum cluster size constraint. The process is modeled using a high-dimensional master equation for the probabilities of cluster configurations. For certain realizations of total mass and maximum cluster sizes, we find exact analytical results for the expected equilibrium cluster distributions. If coagulation is fast relative to fragmentation and if the total system mass is indivisible by the mass of the largest allowed cluster, we find a mean cluster-size distribution that is strikingly broader than that predicted by the corresponding mass-action equations. Combinations ofmore » total mass and maximum cluster size under which equilibration is accelerated, eluding late-stage coarsening, are also delineated. Finally, we compute the mean time it takes particles to first assemble into a maximum-sized cluster. Through careful state-space enumeration, the scaling of mean assembly times is derived for all combinations of total mass and maximum cluster size. We find that CF accelerates assembly relative to monomer kinetic only in special cases. All of our results hold in the infinite system limit and can be only derived from a high-dimensional discrete stochastic model, highlighting how classical mass-action models of self-assembly can fail.« less

A Holistic Multi Evidence Approach to Study the Fragmentation Behaviour of Crystalline Mannitol

PubMed Central

Koner, Jasdip S.; Rajabi-Siahboomi, Ali; Bowen, James; Perrie, Yvonne; Kirby, Daniel; Mohammed, Afzal R.

2015-01-01

Mannitol is an essential excipient employed in orally disintegrating tablets due to its high palatability. However its fundamental disadvantage is its fragmentation during direct compression, producing mechanically weak tablets. The primary aim of this study was to assess the fracture behaviour of crystalline mannitol in relation to the energy input during direct compression, utilising ball milling as the method of energy input, whilst assessing tablet characteristics of post-milled powders. Results indicated that crystalline mannitol fractured at the hydrophilic (011) plane, as observed through SEM, alongside a reduction in dispersive surface energy. Disintegration times of post-milled tablets were reduced due to the exposure of the hydrophilic plane, whilst more robust tablets were produced. This was shown through higher tablet hardness and increased plastic deformation profiles of the post-milled powders, as observed with a lower yield pressure through an out-of-die Heckel analysis. Evaluation of crystal state using x-ray diffraction/differential scanning calorimetry showed that mannitol predominantly retained the β-polymorph; however x-ray diffraction provided a novel method to calculate energy input into the powders during ball milling. It can be concluded that particle size reduction is a pragmatic strategy to overcome the current limitation of mannitol fragmentation and provide improvements in tablet properties. PMID:26553127

Bivalent monoclonal IgY antibody formats by conversion of recombinant antibody fragments.

PubMed

Greunke, Kerstin; Spillner, Edzard; Braren, Ingke; Seismann, Henning; Kainz, Sabine; Hahn, Ulrich; Grunwald, Thomas; Bredehorst, Reinhard

2006-07-13

Monoclonal IgY have the potential to become unique tools for diagnostic research and therapeutic purposes since avian antibodies provide several advantages due to their phylogenetic difference when compared to mammalian antibodies. The mechanism of avian immunoglobulin gene diversification renders chicken an excellent source for the generation of recombinant scFv as well as Fab antibody libraries of high diversity. One major limitation of these antibody fragments, however, is their monovalent format, impairing the functional affinity of the molecules and, thereby, their applicability in prevalent laboratory methods. In this study, we generated vectors for conversion of avian recombinant antibody fragments into different types of bivalent IgY antibody formats. To combine the properties of established mammalian monoclonal antibodies with those of IgY constant domains, we additionally generated bivalent murine/avian chimeric antibody constructs. When expressed in HEK-293 cells, all constructs yielded bivalent disulfide-linked antibodies, which exhibit a glycosylation pattern similar to that of native IgY as assessed by lectin blot analysis. After purification by one step procedures, the chimeric and the entire avian bivalent antibody formats were analyzed for antigen binding and interaction with secondary reagents. The data demonstrate that all antibody formats provide comparable antigen binding characteristics and the well established properties of avian constant domains.

A Holistic Multi Evidence Approach to Study the Fragmentation Behaviour of Crystalline Mannitol

NASA Astrophysics Data System (ADS)

Koner, Jasdip S.; Rajabi-Siahboomi, Ali; Bowen, James; Perrie, Yvonne; Kirby, Daniel; Mohammed, Afzal R.

2015-11-01

Mannitol is an essential excipient employed in orally disintegrating tablets due to its high palatability. However its fundamental disadvantage is its fragmentation during direct compression, producing mechanically weak tablets. The primary aim of this study was to assess the fracture behaviour of crystalline mannitol in relation to the energy input during direct compression, utilising ball milling as the method of energy input, whilst assessing tablet characteristics of post-milled powders. Results indicated that crystalline mannitol fractured at the hydrophilic (011) plane, as observed through SEM, alongside a reduction in dispersive surface energy. Disintegration times of post-milled tablets were reduced due to the exposure of the hydrophilic plane, whilst more robust tablets were produced. This was shown through higher tablet hardness and increased plastic deformation profiles of the post-milled powders, as observed with a lower yield pressure through an out-of-die Heckel analysis. Evaluation of crystal state using x-ray diffraction/differential scanning calorimetry showed that mannitol predominantly retained the β-polymorph however x-ray diffraction provided a novel method to calculate energy input into the powders during ball milling. It can be concluded that particle size reduction is a pragmatic strategy to overcome the current limitation of mannitol fragmentation and provide improvements in tablet properties.

Diverse Metals and Sulfides in Polymict Ureilites EET 83309 and EET 87720

NASA Technical Reports Server (NTRS)

Herrin, J. S.; Mittlefehldt, D. W.; Downes, H.; Humayun, M.

2007-01-01

Ureilites are a group of carbon-bearing ultramafic achondrites. The majority of samples are monomict with major and trace element compositions consistent with a restitic origin after extensive loss of basaltic melts and significant loss of their metallic component during anatexis. Monomict ureilites are thought to represent largely intact samples of the ureilite parent body (UPB) mantle. Polymict ureilites, by contrast, are fragmental breccias consisting of welded lithic clasts and isolated mineral fragments thought to be regolith that assembled after major disruption fragmented large portions of the UPB mantle. In most polymict ureilites, the majority of clasts consist of material similar to monomict ureilites gardened from the UPB mantle but other materials, both endogenic and xenogenic to the UPB are also found in polymict ureilites, including clasts texturally and compositionally similar to known chondrite types as well as feldspathic melt rocks and clasts of Ca-Al-Ti-rich assemblages. In this study, we demonstrate that polymict ureilites also contain a variety of metal and sulfide compositions of diverse origins. They offer insight into the final equilibrium conditions of disrupted portions of the UPB mantle and the diversity of materials locally available for regolith formation, and provide evidence for only limited post-regolith formation thermal metamorphism.

Multi-Homologous Recombination-Based Gene Manipulation in the Rice Pathogen Fusarium fujikuroi

PubMed Central

Hwang, In Sun; Ahn, Il-Pyung

2016-01-01

Gene disruption by homologous recombination is widely used to investigate and analyze the function of genes in Fusarium fujikuroi, a fungus that causes bakanae disease and root rot symptoms in rice. To generate gene deletion constructs, the use of conventional cloning methods, which rely on restriction enzymes and ligases, has had limited success due to a lack of unique restriction enzyme sites. Although strategies that avoid the use of restriction enzymes have been employed to overcome this issue, these methods require complicated PCR steps or are frequently inefficient. Here, we introduce a cloning system that utilizes multi-fragment assembly by In-Fusion to generate a gene disruption construct. This method utilizes DNA fragment fusion and requires only one PCR step and one reaction for construction. Using this strategy, a gene disruption construct for Fusarium cyclin C1 (FCC1 ), which is associated with fumonisin B1 biosynthesis, was successfully created and used for fungal transformation. In vivo and in vitro experiments using confirmed fcc1 mutants suggest that fumonisin production is closely related to disease symptoms exhibited by F. fujikuroi strain B14. Taken together, this multi-fragment assembly method represents a simpler and a more convenient process for targeted gene disruption in fungi. PMID:27298592

Bohemian circular structure, Czechoslovakia: Search for the impact evidence

NASA Technical Reports Server (NTRS)

Rajlich, Petr

1992-01-01

Test of the impact hypothesis for the origin of the circular, 260-km-diameter structure of the Bohemian Massif led to the discovery of glasses and breccias in the Upper Proterozoic sequence that can be compared to autogeneous breccias of larger craters. The black recrystallized glass contains small exsolution crystals of albite-oligoclase and biotite, regularly dispersed in the matrix recrystallized to quartz. The occurrence of these rocks is limited to a 1-sq-km area. It is directly underlain by the breccia of the pelitic and silty rocks cemented by the melted matrix, found on several tens of square kilometers. The melt has the same chemistry as rock fragments in major and in trace elements. It is slightly impoverished in water. The proportion of melted rocks to fragments varies from 1:5 to 10:1. The mineralogy of melt viens is the function of later, mostly contact metamorphism. On the contact of granitic plutons it abounds on sillimanite, cordierite, and small bullets of ilmenite. Immediately on the contact with syenodiorites it contains garnets. The metamorphism of the impact rock melt seems the most probable explanation of the mineralogy and the dry total fusion of rocks accompanied by the strong fragmentation. Other aspects of this investigation are discussed.

Simultaneous ESI-APCI+ ionization and fragmentation pathways for nine benzodiazepines and zolpidem using single quadrupole LC-MS.

PubMed

Galaon, Toma; Vacaresteanu, Catalina; Anghel, Dan-Florin; David, Victor

2014-05-01

Nine important 1,4-benzodiazepines and zolpidem were characterized by liquid chromatography-mass spectrometry using a multimode ionization source able to generate ions using both electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI), and a single quadrupole mass analyzer. An optimum chromatographic separation was applied for all target compounds in less than 8 minutes using a Zorbax Eclipse Plus column (100 × 4.6 mm, 3.5 µm) kept at 35°C and a 0.3% HCOOH/ACN/IPA (61:34:5) mobile phase pumped at 1 ml/min. Optimization of LC-MS method generated low limit of quantitation (LOQ) values situated in the range 0.3-20.5 ng/ml. Comparison between differences in method sensitivity, under specified chromatographic conditions, when using ESI-only, APCI-only, and simultaneous ESI-APCI ionization with such a multimode source was discussed. Mixed ESI-APCI(+) mode proved to be the most sensitive ionization generating an average 35% detector response increase compared to ESI-only ionization and 350% detector response increase with respect to APCI-only ionization. Characterization of the nine benzodiazepines and zolpidem concerning their MS fragmentation pathway following 'in-source' collision-induced dissociation is discussed in detail and some general trends regarding these fragmentations are set. Copyright © 2013 John Wiley & Sons, Ltd.

Demonstration of Laser Induced Acoustic Desoprtion - Chemical Ionization Mass Spectrometry (LIAD-CIMS) for Fragment-Free Measurements of Organic Aerosol Molecular Composition

NASA Astrophysics Data System (ADS)

Browne, E. C.; Abdelhamid, A.; Berry, J.; Alton, M.

2017-12-01

Organic compounds account for a significant portion of fine atmospheric aerosol. Current analytical techniques have provided insights on organic aerosol (OA) sources, composition, and chemical modification pathways. Despite this knowledge, large uncertainties remain and hinder our understanding of aerosol impacts on climate, air quality, and health. Measuring OA composition is challenging due to the complex chemical composition and the wide variation in the properties (e.g., vapor pressure, solubility, reactivity) of organic compounds. In many current measurement techniques, the ability to chemically resolve and quantify OA components is complicated by molecular decomposition, matrix effects, and/or preferential ionization mechanisms. Here, we utilize a novel desorption technique, laser induced acoustic desorption (LIAD), that generates fragment-free, neutral gas-phase molecules. We couple LIAD with a high-resolution chemical ionization mass spectrometer (CIMS) to provide molecular composition OA measurements. Through a series of laboratory experiments, we demonstrate the ability of this technique to measure large, thermally labile species without fragmentation/thermal decomposition. We discuss quantification and detection limits of this technique. We compare LIAD-CIMS measurements with thermal desorption-CIMS measurements using off-line measurements of ambient aerosol collected in Boulder, CO. Lastly, we discuss future development for on-line measurements of OA using LIAD-CIMS.

Landscape matrix mediates occupancy dynamics of Neotropical avian insectivores

USGS Publications Warehouse

Kennedy, Christina M.; Campbell Grant, Evan H.; Neel, Maile C.; Fagan, William F.; Marpa, Peter P.

2011-01-01

In addition to patch-level attributes (i.e., area and isolation), the nature of land cover between habitat patches (the matrix) may drive colonization and extinction dynamics in fragmented landscapes. Despite a long-standing recognition of matrix effects in fragmented systems, an understanding of the relative impacts of different types of land cover on patterns and dynamics of species occurrence remains limited. We employed multi-season occupancy models to determine the relative influence of patch area, patch isolation, within-patch vegetation structure, and landscape matrix on occupancy dynamics of nine Neotropical nsectivorous birds in 99 forest patches embedded in four matrix types (agriculture, suburban evelopment, bauxite mining, and forest) in central Jamaica. We found that within-patch vegetation structure and the matrix type between patches were more important than patch area and patch isolation in determining local colonization and local extinction probabilities, and that the effects of patch area, isolation, and vegetation structure on occupancy dynamics tended to be matrix and species dependent. Across the avian community, the landscape matrix influenced local extinction more than local colonization, indicating that extinction processes, rather than movement, likely drive interspecific differences in occupancy dynamics. These findings lend crucial empirical support to the hypothesis that species occupancy dynamics in fragmented systems may depend greatly upon the landscape context.

iT2DMS: a Standard-Based Diabetic Disease Data Repository and its Pilot Experiment on Diabetic Retinopathy Phenotyping and Examination Results Integration.

PubMed

Wu, Huiqun; Wei, Yufang; Shang, Yujuan; Shi, Wei; Wang, Lei; Li, Jingjing; Sang, Aimin; Shi, Lili; Jiang, Kui; Dong, Jiancheng

2018-06-06

Type 2 diabetes mellitus (T2DM) is a common chronic disease, and the fragment data collected through separated vendors makes continuous management of DM patients difficult. The lack of standard of fragment data from those diabetic patients also makes the further potential phenotyping based on the diabetic data difficult. Traditional T2DM data repository only supports data collection from T2DM patients, lack of phenotyping ability and relied on standalone database design, limiting the secondary usage of these valuable data. To solve these issues, we proposed a novel T2DM data repository framework, which was based on standards. This repository can integrate data from various sources. It would be used as a standardized record for further data transfer as well as integration. Phenotyping was conducted based on clinical guidelines with KNIME workflow. To evaluate the phenotyping performance of the proposed system, data was collected from local community by healthcare providers and was then tested using algorithms. The results indicated that the proposed system could detect DR cases with an average accuracy of about 82.8%. Furthermore, these results had the promising potential of addressing fragmented data. The proposed system has integrating and phenotyping abilities, which could be used for diabetes research in future studies.

Bohemian circular structure, Czechoslovakia: Search for the impact evidence

NASA Astrophysics Data System (ADS)

Rajlich, Petr

Test of the impact hypothesis for the origin of the circular, 260-km-diameter structure of the Bohemian Massif led to the discovery of glasses and breccias in the Upper Proterozoic sequence that can be compared to autogeneous breccias of larger craters. The black recrystallized glass contains small exsolution crystals of albite-oligoclase and biotite, regularly dispersed in the matrix recrystallized to quartz. The occurrence of these rocks is limited to a 1-sq-km area. It is directly underlain by the breccia of the pelitic and silty rocks cemented by the melted matrix, found on several tens of square kilometers. The melt has the same chemistry as rock fragments in major and in trace elements. It is slightly impoverished in water. The proportion of melted rocks to fragments varies from 1:5 to 10:1. The mineralogy of melt viens is the function of later, mostly contact metamorphism. On the contact of granitic plutons it abounds on sillimanite, cordierite, and small bullets of ilmenite. Immediately on the contact with syenodiorites it contains garnets. The metamorphism of the impact rock melt seems the most probable explanation of the mineralogy and the dry total fusion of rocks accompanied by the strong fragmentation. Other aspects of this investigation are discussed.

PROBABILISTIC PROGRAMMING FOR ADVANCED MACHINE LEARNING (PPAML) DISCRIMINATIVE LEARNING FOR GENERATIVE TASKS (DILIGENT)

DTIC Science & Technology

2017-11-29

Structural connections of the frames (fragments) in the knowledge. We call the fundamental elements of the knowledge a limited number of elements...the result of contracted fundamental research deemed exempt from public affairs security and policy review in accordance with SAF/AQR memorandum dated...AVAILABILITY STATEMENT Approved for Public Release; Distribution Unlimited. This report is the result of contracted fundamental research deemed exempt from

[Progress of research on genetic engineering antibody and its application in prevention and control of parasitic diseases].

PubMed

Yao, Yuan; Yu, Chuan-xin

2013-08-01

Antibody has extensive application prospects in the biomedical field. The inherent disadvantages of traditional polyclonal antibody and monoclonal antibody limit their application values. The humanized and fragmented antibody remodeling has given a rise to a series of genetic engineered antibody variant. This paper reviews the progress of research on genetic engineering antibody and its application in prevention and control of parasitic diseases.

An overview of some concepts, potentials, issues, and realities of assisted migration for climate change adaptation in forests

Treesearch

Louis R. Iverson; Matthew P. Peters; Stephen Matthews; Anantha Prasad

2013-01-01

The climate has always been changing, but the rapid rate of climate change, as projected by the IPCC (2007) will likely place unique stresses on plant communities. In addition, anthropogenic barriers (e.g., fragmented land use) present a significant modern constraint that will limit the ability of species migration in responses to a changing climate. As such, managers...

Diversity in Genetic In Vivo Methods for Protein-Protein Interaction Studies: from the Yeast Two-Hybrid System to the Mammalian Split-Luciferase System

PubMed Central

Stynen, Bram; Tournu, Hélène; Tavernier, Jan

2012-01-01

Summary: The yeast two-hybrid system pioneered the field of in vivo protein-protein interaction methods and undisputedly gave rise to a palette of ingenious techniques that are constantly pushing further the limits of the original method. Sensitivity and selectivity have improved because of various technical tricks and experimental designs. Here we present an exhaustive overview of the genetic approaches available to study in vivo binary protein interactions, based on two-hybrid and protein fragment complementation assays. These methods have been engineered and employed successfully in microorganisms such as Saccharomyces cerevisiae and Escherichia coli, but also in higher eukaryotes. From single binary pairwise interactions to whole-genome interactome mapping, the self-reassembly concept has been employed widely. Innovative studies report the use of proteins such as ubiquitin, dihydrofolate reductase, and adenylate cyclase as reconstituted reporters. Protein fragment complementation assays have extended the possibilities in protein-protein interaction studies, with technologies that enable spatial and temporal analyses of protein complexes. In addition, one-hybrid and three-hybrid systems have broadened the types of interactions that can be studied and the findings that can be obtained. Applications of these technologies are discussed, together with the advantages and limitations of the available assays. PMID:22688816

A label-free and high-efficient GO-based aptasensor for cancer cells based on cyclic enzymatic signal amplification.

PubMed

Xiao, Kunyi; Liu, Juan; Chen, Hui; Zhang, Song; Kong, Jilie

2017-05-15

A label-free and high-efficient graphene oxide (GO)-based aptasensor was developed for the detection of low quantity cancer cells based on cell-triggered cyclic enzymatic signal amplification (CTCESA). In the absence of target cells, hairpin aptamer probes (HAPs) and dye-labeled linker DNAs stably coexisted in solution, and the fluorescence was quenched by the GO-based FÖrster resonance energy transfer (FRET) process. In the presence of target cells, the specific binding of HAPs with the target cells triggered a conformational alternation, which resulted in linker DNA complementary pairing and cleavage by nicking endonuclease-strand scission cycles. Consequently, more cleaved fragments of linker DNAs with more the terminal labeled dyes could show the enhanced fluorescence because these cleaved DNA fragments hardly combine with GOs and prevent the FRET process. Fluorescence analysis demonstrated that this GO-based aptasensor exhibited selective and sensitive response to the presence of target CCRF-CEM cells in the concentration range from 50 to 10 5 cells. The detection limit of this method was 25 cells, which was approximately 20 times lower than the detection limit of normal fluorescence aptasensors without amplification. With high sensitivity and specificity, it provided a simple and cost-effective approach for early cancer diagnosis. Copyright © 2016 Elsevier B.V. All rights reserved.

Potential Renoprotective Effects of Rosemary and Thyme Against Gentamicin Toxicity in Rats.

PubMed

Abdel-Azeem, Amal S; Hegazy, Amany M; Zeidan, Hala M; Ibrahim, Khadiga S; El-Sayed, Eman M

2017-07-04

Gentamicin (GM) is a drug used commonly against gram-negative bacteria. Its therapeutic use is mainly limited by its nephrotoxicity. This investigation was aimed at evaluating the nephroprotective effects of rosemary (RM; Rosmarinus officinalis) and thyme (TV; Thymus vulgaris) against GM toxicity. Rats were divided into four groups. Normal control group was treated intraperitoneally (i.p.) with saline; positive control group was administered GM 60 mg/kg/day i.p.; RM group was co-administered 8% RM aqueous extract, 10 mL/kg/day, orally with GM; and TV group was co-administered 8% TV aqueous extract orally, 10 mL/kg/day with GM. Both RM and TV groups extracted significantly ameliorated plasma kidney function biomarkers, and reduced malondialdhyde and glucose levels. Meanwhile, RM extract significantly modulated the electrolyte concentration and increased plasma insulin levels as compared with the GM group. However, the aqueous extract of RM showed more pronounced effects than TV extract. Our data were confirmed by histopathological examination and deoxyribonucleic acid (DNA) fragmentation analysis. Deleterious histopathological changes and increased DNA fragmentation induced by GM treatment were markedly controlled by the co-administration of RM and TV. Such renoprotective influence of RM and TV suggests their concurrent supplementation with GM therapy to limit GM toxicity.

Digital droplet multiple displacement amplification (ddMDA) for whole genome sequencing of limited DNA samples

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rhee, Minsoung; Light, Yooli K.; Meagher, Robert J.

Here, multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently,more » the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per droplet), ddMDA achieves a 65-fold increase in coverage in de novo assembly, and more than 20-fold increase in specificity (percentage of reads mapping to E. coli) compared to the conventional tube MDA. ddMDA offers a powerful method useful for many applications including medical diagnostics, forensics, and environmental microbiology.« less

Scenarios and methods that induce protruding or released CNTs after degradation of nanocomposite materials.

PubMed

Hirth, Sabine; Cena, Lorenzo; Cox, Gerhard; Tomović, Zeljko; Peters, Thomas; Wohlleben, Wendel

2013-04-01

Nanocomposite materials may be considered as a low-risk application of nanotechnology, if the nanofillers remain embedded throughout the life-cycle of the products in which they are embedded. We hypothesize that release of free CNTs occurs by a combination of mechanical stress and chemical degradation of the polymer matrix. We experimentally address limiting cases: Mechanically released fragments may show tubular protrusions on their surface. Here we identify these protrusions unambiguously as naked CNTs by chemically resolved microscopy and a suitable preparation protocol. By size-selective quantification of fragments we establish as a lower limit that at least 95 % of the CNTs remain embedded. Contrary to classical fiber composite approaches, we link this phenomenon to matrix materials with only a few percent elongation at break, predicting which materials should still cover their CNT nanofillers after machining. Protruding networks of CNTs remain after photochemical degradation of the matrix, and we show that it takes the worst case combinations of weathering plus high-shear wear to release free CNTs in the order of mg/m 2 /year. Synergy of chemical degradation and mechanical energy input is identified as the priority scenario of CNT release, but its lab simulation by combined methods is still far from real-world validation.

Digital droplet multiple displacement amplification (ddMDA) for whole genome sequencing of limited DNA samples

DOE PAGES

Rhee, Minsoung; Light, Yooli K.; Meagher, Robert J.; ...

2016-05-04

Here, multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently,more » the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per droplet), ddMDA achieves a 65-fold increase in coverage in de novo assembly, and more than 20-fold increase in specificity (percentage of reads mapping to E. coli) compared to the conventional tube MDA. ddMDA offers a powerful method useful for many applications including medical diagnostics, forensics, and environmental microbiology.« less

High genetic differentiation among French populations of the Orsini's viper (Vipera ursinii ursinii) based on mitochondrial and microsatellite data: implications for conservation management.

PubMed

Ferchaud, Anne-Laure; Lyet, Arnaud; Cheylan, Marc; Arnal, Véronique; Baron, Jean-Pierre; Montgelard, Claudine; Ursenbacher, Sylvain

2011-01-01

The Orsini's viper (Vipera ursinii) is one of the most threatened snakes in Europe due to its highly fragmented distribution and specific open environment (steppic habitat) requirement. French populations are isolated on top of mountain massifs of the southern Prealps/Alps. Mitochondrial sequences (cytochrome b) and 6 microsatellite loci have been used to estimate the levels of genetic diversity and isolation within and among 11 French fragmented populations (a total of 157 individuals). Eleven cytochrome b haplotypes with a limited divergence were observed (mean divergence between haplotypes: 0.31%). However, we detected considerable genetic differentiation among populations (global F(ST) = 0.76 and 0.26 for mitochondrial and nuclear DNA, respectively). Results indicate that 3 populations possibly went through a bottleneck and 1 population showed low genetic diversity compared with the others. Although a significant isolation by distance was detected for both markers, strong differentiation was also observed between geographically close populations, probably due to the ragged landscape that constitutes a serious barrier to gene flow owing to the limited dispersal capability of the viper. Despite some discrepancies between the 2 markers, 8 Management Units have been identified and should be considered for future management projects.

Typing single polymorphic nucleotides in mitochondrial DNA as a way to access Middle Pleistocene DNA

PubMed Central

Valdiosera, Cristina; García, Nuria; Dalén, Love; Smith, Colin; Kahlke, Ralf-Dietrich; Lidén, Kerstin; Angerbjörn, Anders; Arsuaga, Juan Luis; Götherström, Anders

2006-01-01

In this study, we have used a technique designed to target short fragments containing informative mitochondrial substitutions to extend the temporal limits of DNA recovery and study the molecular phylogeny of Ursus deningeri. We present a cladistic analysis using DNA recovered from 400 kyr old U. deningeri remains, which demonstrates U. deningeri's relation to Ursus spelaeus. This study extends the limits of recovery from skeletal remains by almost 300 kyr. Plant material from permafrost environments has yielded DNA of this age in earlier studies, and our data suggest that DNA in teeth from cave environments may be equally well preserved. PMID:17148299

Limited genetic diversity of Brucella spp.

PubMed

Gándara, B; Merino, A L; Rogel, M A; Martínez-Romero, E

2001-01-01

Multilocus enzyme electrophoresis (MLEE) of 99 Brucella isolates, including the type strains from all recognized species, revealed a very limited genetic diversity and supports the proposal of a monospecific genus. In MLEE-derived dendrograms, Brucella abortus and a marine Brucella sp. grouped into a single electrophoretic type related to Brucella neotomae and Brucella ovis. Brucella suis and Brucella canis formed another cluster linked to Brucella melitensis and related to Rhizobium tropici. The Brucella strains tested that were representatives of the six electrophoretic types had the same rRNA gene restriction fragment length polymorphism patterns and identical ribotypes. All 99 isolates had similar chromosome profiles as revealed by the Eckhardt procedure.

Limited Genetic Diversity of Brucella spp.

PubMed Central

Gándara, Benjamín; Merino, Ahidé López; Rogel, Marco Antonio; Martínez-Romero, Esperanza

2001-01-01

Multilocus enzyme electrophoresis (MLEE) of 99 Brucella isolates, including the type strains from all recognized species, revealed a very limited genetic diversity and supports the proposal of a monospecific genus. In MLEE-derived dendrograms, Brucella abortus and a marine Brucella sp. grouped into a single electrophoretic type related to Brucella neotomae and Brucella ovis. Brucella suis and Brucella canis formed another cluster linked to Brucella melitensis and related to Rhizobium tropici. The Brucella strains tested that were representatives of the six electrophoretic types had the same rRNA gene restriction fragment length polymorphism patterns and identical ribotypes. All 99 isolates had similar chromosome profiles as revealed by the Eckhardt procedure. PMID:11136777

Post-Newtonian Jeans Analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nazari, Elham; Kazemi, Ali; Roshan, Mahmood

The Jeans analysis is studied in the first post-Newtonian limit. In other words, the relativistic effects on local gravitational instability are considered for systems whose characteristic velocities and corresponding gravitational fields are higher than those permitted in the Newtonian limit. The dispersion relation for the propagation of small perturbations is found in the post-Newtonian approximation using two different techniques. A new Jeans mass is derived and compared to the standard Jeans mass. In this limit, the relativistic effects make the new Jeans mass smaller than the Newtonian Jeans mass. Furthermore, the fractional difference between these two masses increases when themore » temperature/pressure of the system increases. Interestingly, in this limit, pressure can enhance gravitational instability instead of preventing it. Finally, the results are applied to high-temperature astrophysical systems, and the possibility of local fragmentation in some relativistic systems is investigated.« less

Exploring the limits of ultrafast polymerase chain reaction using liquid for thermal heat exchange: A proof of principle

NASA Astrophysics Data System (ADS)

Maltezos, George; Johnston, Matthew; Taganov, Konstantin; Srichantaratsamee, Chutatip; Gorman, John; Baltimore, David; Chantratita, Wasun; Scherer, Axel

2010-12-01

Thermal ramp rate is a major limiting factor in using real-time polymerase chain reaction (PCR) for routine diagnostics. We explored the limits of speed by using liquid for thermal exchange rather than metal as in traditional devices, and by testing different polymerases. In a clinical setting, our system equaled or surpassed state-of-the-art devices for accuracy in amplifying DNA/RNA of avian influenza, cytomegalovirus, and human immunodeficiency virus. Using Thermococcus kodakaraensis polymerase and optimizing both electrical and chemical systems, we obtained an accurate, 35 cycle amplification of an 85-base pair fragment of E. coli O157:H7 Shiga toxin gene in as little as 94.1 s, a significant improvement over a typical 1 h PCR amplification.

Replicated landscape genetic and network analyses reveal wide variation in functional connectivity for American pikas.

PubMed

Castillo, Jessica A; Epps, Clinton W; Jeffress, Mackenzie R; Ray, Chris; Rodhouse, Thomas J; Schwalm, Donelle

2016-09-01

Landscape connectivity is essential for maintaining viable populations, particularly for species restricted to fragmented habitats or naturally arrayed in metapopulations and facing rapid climate change. The importance of assessing both structural connectivity (physical distribution of favorable habitat patches) and functional connectivity (how species move among habitat patches) for managing such species is well understood. However, the degree to which functional connectivity for a species varies among landscapes, and the resulting implications for conservation, have rarely been assessed. We used a landscape genetics approach to evaluate resistance to gene flow and, thus, to determine how landscape and climate-related variables influence gene flow for American pikas (Ochotona princeps) in eight federally managed sites in the western United States. We used empirically derived, individual-based landscape resistance models in conjunction with predictive occupancy models to generate patch-based network models describing functional landscape connectivity. Metareplication across landscapes enabled identification of limiting factors for dispersal that would not otherwise have been apparent. Despite the cool microclimates characteristic of pika habitat, south-facing aspects consistently represented higher resistance to movement, supporting the previous hypothesis that exposure to relatively high temperatures may limit dispersal in American pikas. We found that other barriers to dispersal included areas with a high degree of topographic relief, such as cliffs and ravines, as well as streams and distances greater than 1-4 km depending on the site. Using the empirically derived network models of habitat patch connectivity, we identified habitat patches that were likely disproportionately important for maintaining functional connectivity, areas in which habitat appeared fragmented, and locations that could be targeted for management actions to improve functional connectivity. We concluded that climate change, besides influencing patch occupancy as predicted by other studies, may alter landscape resistance for pikas, thereby influencing functional connectivity through multiple pathways simultaneously. Spatial autocorrelation among genotypes varied across study sites and was largest where habitat was most dispersed, suggesting that dispersal distances increased with habitat fragmentation, up to a point. This study demonstrates how landscape features linked to climate can affect functional connectivity for species with naturally fragmented distributions, and reinforces the importance of replicating studies across landscapes. © 2016 by the Ecological Society of America.

Observations and analysis of organic aerosol evolution in some prescribed fire smoke plumes

NASA Astrophysics Data System (ADS)

May, A. A.; Lee, T.; McMeeking, G. R.; Akagi, S.; Sullivan, A. P.; Urbanski, S.; Yokelson, R. J.; Kreidenweis, S. M.

2015-06-01

Open biomass burning is a significant source of primary air pollutants such as particulate matter (PM) and non-methane organic gases (NMOG). However, the physical and chemical atmospheric processing of these emissions during transport is poorly understood. Atmospheric transformations of biomass burning emissions have been investigated in environmental chambers, but there have been limited opportunities to investigate these transformations in the atmosphere. In this study, we deployed a suite of real-time instrumentation on a Twin Otter aircraft to sample smoke from prescribed fires in South Carolina, conducting measurements at both the source and downwind to characterize smoke evolution with atmospheric aging. Organic aerosol (OA) within the smoke plumes was quantified using an aerosol mass spectrometer (AMS); refractory black carbon (rBC) was quantified using a single-particle soot photometer, and carbon monoxide (CO) and carbon dioxide (CO2) were measured using a cavity ring-down spectrometer. During the two fires for which we were able to obtain aerosol aging data, normalized excess mixing ratios and "export factors" of conserved species (rBC, CO, CO2) suggested that changes in emissions at the source did not account for most of the differences observed in samples of increasing age. An investigation of AMS mass fragments indicated that the in-plume fractional contribution (fm/z) to OA of the primary fragment (m/z 60) decreased downwind, while the fractional contribution of the secondary fragment (m/z 44) increased. Increases in f44 are typically interpreted as indicating chemical aging of OA. Likewise, we observed an increase in the O : C elemental ratio downwind, which is usually associated with aerosol aging. However, the rapid mixing of these plumes into the background air suggests that these chemical transformations may be attributable to the different volatilities of the compounds that fragment to these m/z in the AMS. The gas-particle partitioning behavior of the bulk OA observed during the study was consistent with the predictions from a parameterization developed for open biomass burning emissions in the laboratory. Furthermore, we observed no statistically significant increase in total organic mass with atmospheric transport. Hence, our results suggest that dilution-driven evaporation likely dominated over the chemical production of secondary organic aerosol (SOA) within our smoke plumes, presumably due to the fast dilution and limited aging times (< ~ 5 h) that we could sample.

Estimating the Grain Size Distribution of Mars based on Fragmentation Theory and Observations

NASA Astrophysics Data System (ADS)

Charalambous, C.; Pike, W. T.; Golombek, M.

2017-12-01

We present here a fundamental extension to the fragmentation theory [1] which yields estimates of the distribution of particle sizes of a planetary surface. The model is valid within the size regimes of surfaces whose genesis is best reflected by the evolution of fragmentation phenomena governed by either the process of meteoritic impacts, or by a mixture with aeolian transportation at the smaller sizes. The key parameter of the model, the regolith maturity index, can be estimated as an average of that observed at a local site using cratering size-frequency measurements, orbital and surface image-detected rock counts and observations of sub-mm particles at landing sites. Through validation of ground truth from previous landed missions, the basis of this approach has been used at the InSight landing ellipse on Mars to extrapolate rock size distributions in HiRISE images down to 5 cm rock size, both to determine the landing safety risk and the subsequent probability of obstruction by a rock of the deployed heat flow mole down to 3-5 m depth [2]. Here we focus on a continuous extrapolation down to 600 µm coarse sand particles, the upper size limit that may be present through aeolian processes [3]. The parameters of the model are first derived for the fragmentation process that has produced the observable rocks via meteorite impacts over time, and therefore extrapolation into a size regime that is affected by aeolian processes has limited justification without further refinement. Incorporating thermal inertia estimates, size distributions observed by the Spirit and Opportunity Microscopic Imager [4] and Atomic Force and Optical Microscopy from the Phoenix Lander [5], the model's parameters in combination with synthesis methods are quantitatively refined further to allow transition within the aeolian transportation size regime. In addition, due to the nature of the model emerging in fractional mass abundance, the percentage of material by volume or mass that resides within the transported fraction on Mars can be estimated. The parameters of the model thus allow for a better understanding of the regolith's history which has implications to the origin of sand on Mars. [1] Charalambous, PhD thesis, ICL, 2015 [2] Golombek et al., Space Science Reviews, 2016 [3] Kok et al., ROPP, 2012 [4] McGlynn et al., JGR, 2011 [5] Pike et al., GRL, 2011

Sudbury project (University of Muenster-Ontario Geological Survey): Sr-Nd in heterolithic breccias and gabbroic dikes

NASA Technical Reports Server (NTRS)

Buhl, D.; Deutsch, A.; Lakomy, R.; Brockmeyer, P.; Dressler, B.

1992-01-01

One major objective of our Sudbury project was to define origin and age of the huge breccia units below and above the Sudbury Igneous Complex (SIC). The heterolithic Footwall Breccia (FB) represents a part of the uplifted crater floor. It contains subrounded fragments up to several meters in size and lithic fragments with shock features (greater than 10 GPa) embedded into a fine- to medium-grained matrix. Epsilon(sub Nd)-epsilon(sub Sr) relationships point to almost exclusively parautochthonous precursor lithologies. The different textures of the matrix reflect the metamorphic history of the breccia layer; thermal annealing by the overlying hot impact melt sheet (SIC) at temperatures greater than 1000 C resulted in melting of the fine crushed material, followed by an episode of metasomatic K-feldspar growth and, finally, formation of low-grade minerals such as actinolite and chlorite. Isotope relationships in the Onaping breccias (Gray and Green Member) are much more complex. All attempts to date the breccia formation failed: Zircons are entirely derived from country rocks and lack the pronounced Pb loss caused by the heat of the slowly cooling impact melt sheet (SIC). Rb-Sr techniques using either lithic fragments of different shock stages or the thin slab method, set time limits for the apparently pervasive alkali mobility in these suevitic breccias. The data array and the intercept in the plots point to a major Rb-Sr fractionation around 1.54 Ga ago. This model age is in the same range as the age obtained for the metasomatic matrix of the FB. Rb-Sr dating of a shock event in impact-related breccias seems to be possible only if their matrix had suffered total melting by the hot melt sheet (FB) or if they contain a high fraction of impact melt (suevitic Onaping breccias), whereas the degree of shock metamorphism in rock or lithic fragments plays a minor role. In the Sudbury case, however, the impact melt in the seuvitic breccias is devitrified and recrystallized, which changed Rb/Sr ratios quite drastically. Therefore, the Onaping breccias give only age limits for alteration and low-grade metamorphism. The Sm-Nd system was not reset during the Sudbury event; clasts as well as the matrix in the FB and in the Onaping breccias show preimpact 'Archean' Nd isotope signatures.

Gene doctoring: a method for recombineering in laboratory and pathogenic Escherichia coli strains.

PubMed

Lee, David J; Bingle, Lewis E H; Heurlier, Karin; Pallen, Mark J; Penn, Charles W; Busby, Stephen J W; Hobman, Jon L

2009-12-09

Homologous recombination mediated by the lambda-Red genes is a common method for making chromosomal modifications in Escherichia coli. Several protocols have been developed that differ in the mechanisms by which DNA, carrying regions homologous to the chromosome, are delivered into the cell. A common technique is to electroporate linear DNA fragments into cells. Alternatively, DNA fragments are generated in vivo by digestion of a donor plasmid with a nuclease that does not cleave the host genome. In both cases the lambda-Red gene products recombine homologous regions carried on the linear DNA fragments with the chromosome. We have successfully used both techniques to generate chromosomal mutations in E. coli K-12 strains. However, we have had limited success with these lambda-Red based recombination techniques in pathogenic E. coli strains, which has led us to develop an enhanced protocol for recombineering in such strains. Our goal was to develop a high-throughput recombineering system, primarily for the coupling of genes to epitope tags, which could also be used for deletion of genes in both pathogenic and K-12 E. coli strains. To that end we have designed a series of donor plasmids for use with the lambda-Red recombination system, which when cleaved in vivo by the I-SceI meganuclease generate a discrete linear DNA fragment, allowing for C-terminal tagging of chromosomal genes with a 6xHis, 3xFLAG, 4xProteinA or GFP tag or for the deletion of chromosomal regions. We have enhanced existing protocols and technologies by inclusion of a cassette conferring kanamycin resistance and, crucially, by including the sacB gene on the donor plasmid, so that all but true recombinants are counter-selected on kanamycin and sucrose containing media, thus eliminating the need for extensive screening. This method has the added advantage of limiting the exposure of cells to the potential damaging effects of the lambda-Red system, which can lead to unwanted secondary alterations to the chromosome. We have developed a counter-selective recombineering technique for epitope tagging or for deleting genes in E. coli. We have demonstrated the versatility of the technique by modifying the chromosome of the enterohaemorrhagic O157:H7 (EHEC), uropathogenic CFT073 (UPEC), enteroaggregative O42 (EAEC) and enterotoxigenic H10407 (ETEC) E. coli strains as well as in K-12 laboratory strains.