Prediction of hot spots in protein interfaces using a random forest model with hybrid features.

Science.gov (United States)

Wang, Lin; Liu, Zhi-Ping; Zhang, Xiang-Sun; Chen, Luonan

2012-03-01

Prediction of hot spots in protein interfaces provides crucial information for the research on protein-protein interaction and drug design. Existing machine learning methods generally judge whether a given residue is likely to be a hot spot by extracting features only from the target residue. However, hot spots usually form a small cluster of residues which are tightly packed together at the center of protein interface. With this in mind, we present a novel method to extract hybrid features which incorporate a wide range of information of the target residue and its spatially neighboring residues, i.e. the nearest contact residue in the other face (mirror-contact residue) and the nearest contact residue in the same face (intra-contact residue). We provide a novel random forest (RF) model to effectively integrate these hybrid features for predicting hot spots in protein interfaces. Our method can achieve accuracy (ACC) of 82.4% and Matthew's correlation coefficient (MCC) of 0.482 in Alanine Scanning Energetics Database, and ACC of 77.6% and MCC of 0.429 in Binding Interface Database. In a comparison study, performance of our RF model exceeds other existing methods, such as Robetta, FOLDEF, KFC, KFC2, MINERVA and HotPoint. Of our hybrid features, three physicochemical features of target residues (mass, polarizability and isoelectric point), the relative side-chain accessible surface area and the average depth index of mirror-contact residues are found to be the main discriminative features in hot spots prediction. We also confirm that hot spots tend to form large contact surface areas between two interacting proteins. Source data and code are available at: http://www.aporc.org/doc/wiki/HotSpot.

Multiple proteins of White spot syndrome virus involved in ...

Indian Academy of Sciences (India)

2014-03-20

Mar 20, 2014 ... β-integrin with structure proteins of WSSV and motifs involved in WSSV infection was examined. The results showed ... Introduction. White spot ... denatured conditions and renatured by successive 12 h incu- bations with 6, 4, ...

Blind spot detection & passive lane change assist systems

NARCIS (Netherlands)

Surovtcev, I.

2015-01-01

The project goal was design and implementation of proof-of-concept for two systems that aim to tackle the blind spot problem of for the commercial vehicles: Blind Spot Detection and Passive Lane Change Assist functions. The system implementation was done using Rapid Control Prototype (RCP) hardware.

Dried Blood Spot Proteomics: Surface Extraction of Endogenous Proteins Coupled with Automated Sample Preparation and Mass Spectrometry Analysis

Science.gov (United States)

Martin, Nicholas J.; Bunch, Josephine; Cooper, Helen J.

2013-08-01

Dried blood spots offer many advantages as a sample format including ease and safety of transport and handling. To date, the majority of mass spectrometry analyses of dried blood spots have focused on small molecules or hemoglobin. However, dried blood spots are a potentially rich source of protein biomarkers, an area that has been overlooked. To address this issue, we have applied an untargeted bottom-up proteomics approach to the analysis of dried blood spots. We present an automated and integrated method for extraction of endogenous proteins from the surface of dried blood spots and sample preparation via trypsin digestion by use of the Advion Biosciences Triversa Nanomate robotic platform. Liquid chromatography tandem mass spectrometry of the resulting digests enabled identification of 120 proteins from a single dried blood spot. The proteins identified cross a concentration range of four orders of magnitude. The method is evaluated and the results discussed in terms of the proteins identified and their potential use as biomarkers in screening programs.

Morphological changes in ultrafast laser ablation plumes with varying spot size.

Science.gov (United States)

Harilal, S S; Diwakar, P K; Polek, M P; Phillips, M C

2015-06-15

We investigated the role of spot size on plume morphology during ultrafast laser ablation of metal targets. Our results show that the spatial features of fs LA plumes are strongly dependent on the focal spot size. Two-dimensional self-emission images showed that the shape of the ultrafast laser ablation plumes changes from spherical to cylindrical with an increasing spot size from 100 to 600 μm. The changes in plume morphology and internal structures are related to ion emission dynamics from the plasma, where broader angular ion distribution and faster ions are noticed for the smallest spot size used. The present results clearly show that the morphological changes in the plume with spot size are independent of laser pulse width.

Spot urine protein measurements in normotensive pregnancies, pregnancies with isolated proteinuria and preeclampsia.

Science.gov (United States)

Kattah, Andrea; Milic, Natasa; White, Wendy; Garovic, Vesna

2017-10-01

We performed a prospective, longitudinal study of pregnant women presenting to their first obstetrics visits to characterize the changes in spot urine protein-to-creatinine (UPCR) and albumin-to-creatinine ratios (UACR) in normotensive pregnancies, as well as identify clinical characteristics associated with isolated proteinuria and preeclampsia. We measured spot urinary albumin, protein, and creatinine at the first prenatal visit, end of the second trimester, and at delivery. In the normotensive pregnancies ( n = 142), we found that from the beginning of pregnancy to delivery, UACR increased by a median [interquartile range (IQR)] of 14.7 mg/g Cr (3.74-51.8) and UPCR by 60 mg/g Cr (30-130) ( P 300 mg/g Cr in the absence of hypertension) was identified in 19/142 (13.4%) normotensive pregnancies. Increases in systolic and diastolic blood pressure from early pregnancy to delivery and increases in UACR from early to midpregnancy were associated with isolated proteinuria at delivery. Twelve women developed preeclampsia. Nulliparity, early, and midpregnancy diastolic blood pressures were strongly associated with the development of preeclampsia, but early changes in UACR were not. In conclusion, women who develop isolated proteinuria at delivery have a larger increase in blood pressure than women without proteinuria and have a "microalbuminuric" phase earlier in gestation, unlike women who develop preeclampsia. These findings suggest a different mechanism of urine protein excretion in women with isolated proteinuria as compared with women with preeclampsia, where proteinuria has a more abrupt onset. Copyright © 2017 the American Physiological Society.

Estimation of daily protein intake based on spot urine urea nitrogen concentration in chronic kidney disease patients.

Science.gov (United States)

Kanno, Hiroko; Kanda, Eiichiro; Sato, Asako; Sakamoto, Kaori; Kanno, Yoshihiko

2016-04-01

Determination of daily protein intake in the management of chronic kidney disease (CKD) requires precision. Inaccuracies in recording dietary intake occur, and estimation from total urea excretion presents hurdles owing to the difficulty of collecting whole urine for 24 h. Spot urine has been used for measuring daily sodium intake and urinary protein excretion. In this cross-sectional study, we investigated whether urea nitrogen (UN) concentration in spot urine can be used to predict daily protein intake instead of the 24-h urine collection in 193 Japanese CKD patients (Stages G1-G5). After patient randomization into 2 datasets for the development and validation of models, bootstrapping was used to develop protein intake estimation models. The parameters for the candidate multivariate regression models were male gender, age, body mass index (BMI), diabetes mellitus, dyslipidemia, proteinuria, estimated glomerular filtration rate, serum albumin level, spot urinary UN and creatinine level, and spot urinary UN/creatinine levels. The final model contained BMI and spot urinary UN level. The final model was selected because of the higher correlation between the predicted and measured protein intakes r = 0.558 (95 % confidence interval 0.400, 0.683), and the smaller distribution of the difference between the measured and predicted protein intakes than those of the other models. The results suggest that UN concentration in spot urine may be used to estimate daily protein intake and that a prediction formula would be useful for nutritional control in CKD patients.

CORRELATION OF SPOT URINE ALBUMIN AND 12-HOUR URINE PROTEIN WITH 24-HOUR URINE PROTEIN IN PRE-ECLAMPSIA

Directory of Open Access Journals (Sweden)

S. Vinayachandran

2017-11-01

Full Text Available BACKGROUND Pre-eclampsia is defined as the development of new-onset hypertension in the second half of pregnancy often accompanied by new-onset proteinuria with other signs and symptoms. Proteinuria is defined by the excretion of 300 mg or more of protein in a 24-hour urine collection. To avoid time consumed in collection of 24-hour urine specimens, efforts have been made to develop faster methods to determine concentration of urine protein. Preliminary studies have suggested that 12-hour urine protein collection maybe adequate for evaluation of pre-eclampsia with advantage of early diagnosis and treatment of pre-eclampsia as well as potential for early hospital discharge and increased compliance with specimen collection. The aim of the study is to evaluate and correlate spot urine albumin and 12-hour urine protein with 24-hour urine protein in pre-eclampsia. MATERIALS AND METHODS A diagnostic evaluation study- a 24-hour urine protein, 12-hour urine protein and spot urine albumin results are analysed. Correlation of 12-hour urine protein and spot urine albumin with 24-hour urine protein is analysed using SPSS software. The strength of correlation was measured by Pearson’s correlation coefficient (r. Student’s t-test and Chi-square tests were used to compare patients with and without 24-hour urine protein ≥300 mg. Probability value of 165 mg with 24-hour urine protein ≥300 mg suggest that this test has role in the evaluation of women with suspected pre-eclampsia and could be substituted for 24-hour urine protein as a simple, faster and cheaper method.

DBAC: A simple prediction method for protein binding hot spots based on burial levels and deeply buried atomic contacts

Science.gov (United States)

2011-01-01

Background A protein binding hot spot is a cluster of residues in the interface that are energetically important for the binding of the protein with its interaction partner. Identifying protein binding hot spots can give useful information to protein engineering and drug design, and can also deepen our understanding of protein-protein interaction. These residues are usually buried inside the interface with very low solvent accessible surface area (SASA). Thus SASA is widely used as an outstanding feature in hot spot prediction by many computational methods. However, SASA is not capable of distinguishing slightly buried residues, of which most are non hot spots, and deeply buried ones that are usually inside a hot spot. Results We propose a new descriptor called “burial level” for characterizing residues, atoms and atomic contacts. Specifically, burial level captures the depth the residues are buried. We identify different kinds of deeply buried atomic contacts (DBAC) at different burial levels that are directly broken in alanine substitution. We use their numbers as input for SVM to classify between hot spot or non hot spot residues. We achieve F measure of 0.6237 under the leave-one-out cross-validation on a data set containing 258 mutations. This performance is better than other computational methods. Conclusions Our results show that hot spot residues tend to be deeply buried in the interface, not just having a low SASA value. This indicates that a high burial level is not only a necessary but also a more sufficient condition than a low SASA for a residue to be a hot spot residue. We find that those deeply buried atoms become increasingly more important when their burial levels rise up. This work also confirms the contribution of deeply buried interfacial atomic contacts to the energy of protein binding hot spot. PMID:21689480

Co-Occurring Atomic Contacts for the Characterization of Protein Binding Hot Spots

Science.gov (United States)

Liu, Qian; Ren, Jing; Song, Jiangning; Li, Jinyan

2015-01-01

A binding hot spot is a small area at a protein-protein interface that can make significant contribution to binding free energy. This work investigates the substantial contribution made by some special co-occurring atomic contacts at a binding hot spot. A co-occurring atomic contact is a pair of atomic contacts that are close to each other with no more than three covalent-bond steps. We found that two kinds of co-occurring atomic contacts can play an important part in the accurate prediction of binding hot spot residues. One is the co-occurrence of two nearby hydrogen bonds. For example, mutations of any residue in a hydrogen bond network consisting of multiple co-occurring hydrogen bonds could disrupt the interaction considerably. The other kind of co-occurring atomic contact is the co-occurrence of a hydrophobic carbon contact and a contact between a hydrophobic carbon atom and a π ring. In fact, this co-occurrence signifies the collective effect of hydrophobic contacts. We also found that the B-factor measurements of several specific groups of amino acids are useful for the prediction of hot spots. Taking the B-factor, individual atomic contacts and the co-occurring contacts as features, we developed a new prediction method and thoroughly assessed its performance via cross-validation and independent dataset test. The results show that our method achieves higher prediction performance than well-known methods such as Robetta, FoldX and Hotpoint. We conclude that these contact descriptors, in particular the novel co-occurring atomic contacts, can be used to facilitate accurate and interpretable characterization of protein binding hot spots. PMID:26675422

How many proteins can be identified in a 2DE gel spot within an analysis of a complex human cancer tissue proteome?

Science.gov (United States)

Zhan, Xianquan; Yang, Haiyan; Peng, Fang; Li, Jianglin; Mu, Yun; Long, Ying; Cheng, Tingting; Huang, Yuda; Li, Zhao; Lu, Miaolong; Li, Na; Li, Maoyu; Liu, Jianping; Jungblut, Peter R

2018-04-01

Two-dimensional gel electrophoresis (2DE) in proteomics is traditionally assumed to contain only one or two proteins in each 2DE spot. However, 2DE resolution is being complemented by the rapid development of high sensitivity mass spectrometers. Here we compared MALDI-MS, LC-Q-TOF MS and LC-Orbitrap Velos MS for the identification of proteins within one spot. With LC-Orbitrap Velos MS each Coomassie Blue-stained 2DE spot contained an average of at least 42 and 63 proteins/spot in an analysis of a human glioblastoma proteome and a human pituitary adenoma proteome, respectively, if a single gel spot was analyzed. If a pool of three matched gel spots was analyzed this number further increased up to an average of 230 and 118 proteins/spot for glioblastoma and pituitary adenoma proteome, respectively. Multiple proteins per spot confirm the necessity of isotopic labeling in large-scale quantification of different protein species in a proteome. Furthermore, a protein abundance analysis revealed that most of the identified proteins in each analyzed 2DE spot were low-abundance proteins. Many proteins were present in several of the analyzed spots showing the ability of 2DE-MS to separate at the protein species level. Therefore, 2DE coupled with high-sensitivity LC-MS has a clearly higher sensitivity as expected until now to detect, identify and quantify low abundance proteins in a complex human proteome with an estimated resolution of about 500 000 protein species. This clearly exceeds the resolution power of bottom-up LC-MS investigations. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Computational Prediction of Hot Spot Residues

Science.gov (United States)

Morrow, John Kenneth; Zhang, Shuxing

2013-01-01

Most biological processes involve multiple proteins interacting with each other. It has been recently discovered that certain residues in these protein-protein interactions, which are called hot spots, contribute more significantly to binding affinity than others. Hot spot residues have unique and diverse energetic properties that make them challenging yet important targets in the modulation of protein-protein complexes. Design of therapeutic agents that interact with hot spot residues has proven to be a valid methodology in disrupting unwanted protein-protein interactions. Using biological methods to determine which residues are hot spots can be costly and time consuming. Recent advances in computational approaches to predict hot spots have incorporated a myriad of features, and have shown increasing predictive successes. Here we review the state of knowledge around protein-protein interactions, hot spots, and give an overview of multiple in silico prediction techniques of hot spot residues. PMID:22316154

Multiple proteins of White spot syndrome virus involved in ...

Indian Academy of Sciences (India)

The recognition and attachment of virus to its host cell surface is a critical step for viral infection. Recent research revealed that -integrin was involved in White spot syndrome virus (WSSV) infection. In this study, the interaction of -integrin with structure proteins of WSSV and motifs involved in WSSV infection was ...

Water-Soluble Dried Blood Spot in Protein Analysis: A Proof-of-Concept Study.

Science.gov (United States)

Rosting, Cecilie; Gjelstad, Astrid; Halvorsen, Trine Grønhaug

2015-08-04

In the present work human chorionic gonadotropin (hCG) was used as a model protein in a proof-of-concept study combining water-soluble dried blood spot (DBS) material in liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based protein analysis. A water-soluble material consisting of commercially available carboxymethyl cellulose (CMC) was evaluated as sampling material for this purpose. The material dissolved readily at physiological pH. Different sample preparation methods were evaluated, and in the final method, 15 μL of whole blood was deposited and dried on CMC before the whole spot was dissolved prior to cleanup by immunoaffinity extraction, tryptic digest, and preconcentration by solid-phase extraction (SPE). The results indicated complete dissolution of hCG from the spots, acceptable limit of detection (LOD) (0.1 IU/mL), linearity (R(2) = 0.959), accuracy (16%), and precision (≤22%). Long-term stability (45 days) of hCG in dried spots at reduced temperatures (≤8 °C) was also demonstrated. The analyte recovery was comparable to the commercially available nonsolvable cellulose material (FTA DMPK-C card).

Hot-spot analysis to dissect the functional protein-protein interface of a tRNA-modifying enzyme.

Science.gov (United States)

Jakobi, Stephan; Nguyen, Tran Xuan Phong; Debaene, François; Metz, Alexander; Sanglier-Cianférani, Sarah; Reuter, Klaus; Klebe, Gerhard

2014-10-01

Interference with protein-protein interactions of interfaces larger than 1500 Ų by small drug-like molecules is notoriously difficult, particularly if targeting homodimers. The tRNA modifying enzyme Tgt is only functionally active as a homodimer. Thus, blocking Tgt dimerization is a promising strategy for drug therapy as this protein is key to the development of Shigellosis. Our goal was to identify hot-spot residues which, upon mutation, result in a predominantly monomeric state of Tgt. The detailed understanding of the spatial location and stability contribution of the individual interaction hot-spot residues and the plasticity of motifs involved in the interface formation is a crucial prerequisite for the rational identification of drug-like inhibitors addressing the respective dimerization interface. Using computational analyses, we identified hot-spot residues that contribute particularly to dimer stability: a cluster of hydrophobic and aromatic residues as well as several salt bridges. This in silico prediction led to the identification of a promising double mutant, which was validated experimentally. Native nano-ESI mass spectrometry showed that the dimerization of the suggested mutant is largely prevented resulting in a predominantly monomeric state. Crystal structure analysis and enzyme kinetics of the mutant variant further support the evidence for enhanced monomerization and provide first insights into the structural consequences of the dimer destabilization. © 2014 Wiley Periodicals, Inc.

Protein profile of Beta vulgaris leaf apoplastic fluid and changes induced by Fe deficiency and Fe resupply

Directory of Open Access Journals (Sweden)

Laura eCeballos-Laita

2015-03-01

Full Text Available The fluid collected by direct leaf centrifugation has been used to study the proteome of the sugar beet apoplastic fluid as well as the changes induced by Fe deficiency and Fe resupply to Fe-deficient plants in the protein profile. Plants were grown in Fe-sufficient and Fe-deficient conditions, and Fe resupply was carried out with 45 μM Fe(III-EDTA for 24 h. Protein extracts of leaf apoplastic fluid were analyzed by two-dimensional isoelectric focusing-SDS-PAGE electrophoresis. Gel image analysis revealed 203 consistent spots, and proteins in 81% of them (164 were identified by nLC-MS/MS using a custom made reference repository of beet protein sequences. When redundant UniProt entries were deleted, a non-redundant leaf apoplastic proteome consisting of 109 proteins was obtained. TargetP and SecretomeP algorithms predicted that 63% of them were secretory proteins. Functional classification of the non-redundant proteins indicated that stress and defense, protein metabolism, cell wall and C metabolism accounted for approximately 75% of the identified proteome. The effects of Fe-deficiency on the leaf apoplast proteome were limited, with only five spots (2.5% changing in relative abundance, thus suggesting that protein homeostasis in the leaf apoplast fluid is well maintained upon Fe shortage. The identification of three chitinase isoforms among proteins increasing in relative abundance with Fe-deficiency suggests that one of the few effects of Fe deficiency in the leaf apoplast proteome includes cell wall modifications. Iron resupply to Fe deficient plants changed the relative abundance of 16 spots when compared to either Fe-sufficient or Fe-deficient samples. Proteins identified in these spots can be broadly classified as those responding to Fe-resupply, which included defense and cell wall related proteins, and non-responsive, which are mainly protein metabolism related proteins and whose changes in relative abundance followed the same trend as

Liver spots

Science.gov (United States)

... skin changes - liver spots; Senile or solar lentigines; Skin spots - aging; Age spots ... Liver spots are changes in skin color that occur in older skin. The coloring may be due to aging, exposure to the sun ...

IL-1beta induced protein changes in diabetes prone BB rat islets of Langerhans identified by proteome analysis

DEFF Research Database (Denmark)

Sparre, T; Bjerre-Christensen, Ulla; Mose Larsen, P

2002-01-01

of 82 out of 1 815 protein spots detected by two dimensional gel electrophoresis in IL-1beta exposed diabetes prone Bio Breeding (BB-DP) rat islets of Langerhans in vitro. The aim of this study was to identify the proteins in these 82 spots by mass spectrometry and compare these changes with those seen......Type I (insulin-dependent) diabetes mellitus is characterized by selective destruction of the insulin producing beta cells. Interleukin-1beta (IL-1beta) modulates the beta-cell function, protein synthesis, energy production and causes apoptosis. We have previously shown changes in the expression...

Protein expression profiling by antibody array analysis with use of dried blood spot samples on filter paper.

Science.gov (United States)

Jiang, Weidong; Mao, Ying Qing; Huang, Ruochun; Duan, Chaohui; Xi, Yun; Yang, Kai; Huang, Ruo-Pan

2014-01-31

Dried blood spot samples (DBSS) on filter paper offer several advantages compared to conventional serum/plasma samples: they do not require any phlebotomy or separation of blood by centrifugation; they are less invasive; they allow sample stability and shipment at room temperature; and they pose a negligible risk of infection with blood-borne viruses, such as HIV, HBV and HCV, to those who handle them. Therefore dried blood spot samples (DBSS) on filter paper can be a quick, convenient and inexpensive means of obtaining blood samples for biomarker discovery, disease screening, diagnosis and treatment monitoring in non-hospitalized, public health settings. In this study, we investigated for the first time the potential application of dried blood spot samples (DBSS) in protein expression profiling using antibody array technology. First, optimal conditions for array assay performance using dried blood spot samples (DBSS) was established, including sample elution buffer, elution time, elution temperature and assay blocking buffer. Second, we analyzed dried blood spot samples (DBSS) using three distinct antibody array platforms, including sandwich-based antibody arrays, quantitative antibody arrays and biotin-label-based antibody arrays. In comparison with paired serum samples, detection of circulating proteins in dried blood spot samples (DBSS) correlated well for both low- and high-abundance proteins on all three antibody array platforms. In conclusion, our study strongly indicates the novel application of multiplex antibody array platforms to analyze dried blood spot samples (DBSS) on filter paper represents a viable, cost-effective method for protein profiling, biomarker discovery and disease screening in a large, population-based survey. Copyright © 2013 Elsevier B.V. All rights reserved.

Identification of a new genomic hot spot of evolutionary diversification of protein function.

Directory of Open Access Journals (Sweden)

Aline Winkelmann

Full Text Available Establishment of phylogenetic relationships remains a challenging task because it is based on computational analysis of genomic hot spots that display species-specific sequence variations. Here, we identify a species-specific thymine-to-guanine sequence variation in the Glrb gene which gives rise to species-specific splice donor sites in the Glrb genes of mouse and bushbaby. The resulting splice insert in the receptor for the inhibitory neurotransmitter glycine (GlyR conveys synaptic receptor clustering and specific association with a particular synaptic plasticity-related splice variant of the postsynaptic scaffold protein gephyrin. This study identifies a new genomic hot spot which contributes to phylogenetic diversification of protein function and advances our understanding of phylogenetic relationships.

Estimating mean change in population salt intake using spot urine samples.

Science.gov (United States)

Petersen, Kristina S; Wu, Jason H Y; Webster, Jacqui; Grimes, Carley; Woodward, Mark; Nowson, Caryl A; Neal, Bruce

2017-10-01

Spot urine samples are easier to collect than 24-h urine samples and have been used with estimating equations to derive the mean daily salt intake of a population. Whether equations using data from spot urine samples can also be used to estimate change in mean daily population salt intake over time is unknown. We compared estimates of change in mean daily population salt intake based upon 24-h urine collections with estimates derived using equations based on spot urine samples. Paired and unpaired 24-h urine samples and spot urine samples were collected from individuals in two Australian populations, in 2011 and 2014. Estimates of change in daily mean population salt intake between 2011 and 2014 were obtained directly from the 24-h urine samples and by applying established estimating equations (Kawasaki, Tanaka, Mage, Toft, INTERSALT) to the data from spot urine samples. Differences between 2011 and 2014 were calculated using mixed models. A total of 1000 participants provided a 24-h urine sample and a spot urine sample in 2011, and 1012 did so in 2014 (paired samples n = 870; unpaired samples n = 1142). The participants were community-dwelling individuals living in the State of Victoria or the town of Lithgow in the State of New South Wales, Australia, with a mean age of 55 years in 2011. The mean (95% confidence interval) difference in population salt intake between 2011 and 2014 determined from the 24-h urine samples was -0.48g/day (-0.74 to -0.21; P spot urine samples was -0.24 g/day (-0.42 to -0.06; P = 0.01) using the Tanaka equation, -0.42 g/day (-0.70 to -0.13; p = 0.004) using the Kawasaki equation, -0.51 g/day (-1.00 to -0.01; P = 0.046) using the Mage equation, -0.26 g/day (-0.42 to -0.10; P = 0.001) using the Toft equation, -0.20 g/day (-0.32 to -0.09; P = 0.001) using the INTERSALT equation and -0.27 g/day (-0.39 to -0.15; P  0.058). Separate analysis of the unpaired and paired data showed that detection of

Age-related differences in plasma proteins: how plasma proteins change from neonates to adults.

Directory of Open Access Journals (Sweden)

Vera Ignjatovic

Full Text Available The incidence of major diseases such as cardiovascular disease, thrombosis and cancer increases with age and is the major cause of mortality world-wide, with neonates and children somehow protected from such diseases of ageing. We hypothesized that there are major developmental differences in plasma proteins and that these contribute to age-related changes in the incidence of major diseases. We evaluated the human plasma proteome in healthy neonates, children and adults using the 2D-DIGE approach. We demonstrate significant changes in number and abundance of up to 100 protein spots that have marked differences in during the transition of the plasma proteome from neonate and child through to adult. These proteins are known to be involved in numerous physiological processes such as iron transport and homeostasis, immune response, haemostasis and apoptosis, amongst others. Importantly, we determined that the proteins that are differentially expressed with age are not the same proteins that are differentially expressed with gender and that the degree of phosphorylation of plasma proteins also changes with age. Given the multi-functionality of these proteins in human physiology, understanding the differences in the plasma proteome in neonates and children compared to adults will make a major contribution to our understanding of developmental biology in humans.

Cellular protein receptors of maculosin, a host specific phytotoxin of spotted knapweed (Centaurea maculosa L.).

Science.gov (United States)

Park, S H; Strobel, G A

1994-01-05

Maculosin (the diketopiperazine, cyclo (L-Pro-L-Tyr)) is a host specific phytotoxin produced by Alternaria alternata on spotted knapweed (Centaurea maculosa L.). Receptors for this phytotoxin have been isolated from spotted knapweed. Knapweed leaves possess most of the maculosin-binding activity in the cytosolic fraction. However, activity was also observed in the whole membrane fraction of the leaf. The binding component of the cytosolic fraction was identified as a protein(s) because of its heat-lability and sensitivity to proteases. A 16-fold purification of a toxin-binding protein was carried out by ammonium sulfate fractionation, and Sephadex G-200, and maculosin-affinity column chromatography. The affinity column was prepared with epoxy activated Sepharose 6B to which the phenolic group of maculosin was attached. The receptor was estimated to contain more than one binding protein by native and SDS-PAGE. At least one of the maculosin-binding proteins was identified as ribulose-1,5-biphosphate carboxylase (RuBPcase).

A Machine Learning Approach for Hot-Spot Detection at Protein-Protein Interfaces

Directory of Open Access Journals (Sweden)

Rita Melo

2016-07-01

Full Text Available Understanding protein-protein interactions is a key challenge in biochemistry. In this work, we describe a more accurate methodology to predict Hot-Spots (HS in protein-protein interfaces from their native complex structure compared to previous published Machine Learning (ML techniques. Our model is trained on a large number of complexes and on a significantly larger number of different structural- and evolutionary sequence-based features. In particular, we added interface size, type of interaction between residues at the interface of the complex, number of different types of residues at the interface and the Position-Specific Scoring Matrix (PSSM, for a total of 79 features. We used twenty-seven algorithms from a simple linear-based function to support-vector machine models with different cost functions. The best model was achieved by the use of the conditional inference random forest (c-forest algorithm with a dataset pre-processed by the normalization of features and with up-sampling of the minor class. The method has an overall accuracy of 0.80, an F1-score of 0.73, a sensitivity of 0.76 and a specificity of 0.82 for the independent test set.

The Effect of Thermal Lamination Processes on Colorimetric Change in Spot Colours

Directory of Open Access Journals (Sweden)

Eduard Galić

2015-03-01

Full Text Available Understanding the effect of laminating processes on spot colours is of great importance in the offset printing process, especially given the application versatility of spot colours. Laminating process, as a very common process and one of the first in a sequence of finishing processes in graphics production, can affect print’s visual impression to varying degrees. Spot colours, as mixtures of different ratios of inks, are subject to a change due to matt or gloss lamination process. The research examined the impact of thermal lamination processes on printed spot colours on different printing substrates. The degree of change on prints caused by laminating films in the thermal process was determined using spectrophotometric and densitometric methods. Particular emphasis is placed on the spot colour because of its specific characteristics. Research results are shown in charts and they are showing clearly the modality and the extent laminating processes effect the colorimetric difference in laminated and non-laminated prints. This scientific research provides objective conclusions that help in predicting the possible variations within the usage of laminating processes.

Estimating rice grain protein contents with SPOT/HRV data acquired at maturing stage

International Nuclear Information System (INIS)

Asaka, D.; Shiga, H.

2003-01-01

Rice grain protein contents that play an important role in the eating quality of rice can be estimated from leaf color in maturing stage. In order to investigate the distribution of paddy rice grain protein of a wide area, we employed SPOT/HRV data from August to September for successive 4 years, selecting the Naganuma town, Hokkaido as the study area. The relationship between each spectral bands and ground survey data were examined. The result showed that the grain protein contents could be estimated using the normalized difference vegetation index (NDVI) with the absolute root mean square error less than 0.4% under the condition that the time lag between the satellite observation date and the maturing stage was within 20 days. In this period, we would have enough chance to get clear observation data every year under the weather conditions in the study area using the SPOT/HRV sensors that has pointing ability. For major rice varieties cultivated in Hokkaido, the same relationship between NDVI and protein contents was observed. Thus, we conclude that the method proposed in this study is operational in rice production

Nucleocapsid protein VP15 is the basic DNA binding protein of white spot syndrome virus of shrimp

NARCIS (Netherlands)

Witteveldt, J.; Vermeesch, A.M.G.; Langenhof, M.; Lang, de A.; Vlak, J.M.; Hulten, van M.C.W.

2005-01-01

White spot syndrome virus (WSSV) is type species of the genus Whispovirus of the new family Nimaviridae. Despite the elucidation of its genomic sequence, very little is known about the virus as only 6% of its ORFs show homology to known genes. One of the structural virion proteins, VP15, is part of

Predicting hot spots in protein interfaces based on protrusion index, pseudo hydrophobicity and electron-ion interaction pseudopotential features

Science.gov (United States)

Xia, Junfeng; Yue, Zhenyu; Di, Yunqiang; Zhu, Xiaolei; Zheng, Chun-Hou

2016-01-01

The identification of hot spots, a small subset of protein interfaces that accounts for the majority of binding free energy, is becoming more important for the research of drug design and cancer development. Based on our previous methods (APIS and KFC2), here we proposed a novel hot spot prediction method. For each hot spot residue, we firstly constructed a wide variety of 108 sequence, structural, and neighborhood features to characterize potential hot spot residues, including conventional ones and new one (pseudo hydrophobicity) exploited in this study. We then selected 3 top-ranking features that contribute the most in the classification by a two-step feature selection process consisting of minimal-redundancy-maximal-relevance algorithm and an exhaustive search method. We used support vector machines to build our final prediction model. When testing our model on an independent test set, our method showed the highest F1-score of 0.70 and MCC of 0.46 comparing with the existing state-of-the-art hot spot prediction methods. Our results indicate that these features are more effective than the conventional features considered previously, and that the combination of our and traditional features may support the creation of a discriminative feature set for efficient prediction of hot spots in protein interfaces. PMID:26934646

A 3D model of the membrane protein complex formed by the white spot syndrome virus structural proteins.

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Yun-Shiang Chang

Full Text Available BACKGROUND: Outbreaks of white spot disease have had a large negative economic impact on cultured shrimp worldwide. However, the pathogenesis of the causative virus, WSSV (whit spot syndrome virus, is not yet well understood. WSSV is a large enveloped virus. The WSSV virion has three structural layers surrounding its core DNA: an outer envelope, a tegument and a nucleocapsid. In this study, we investigated the protein-protein interactions of the major WSSV structural proteins, including several envelope and tegument proteins that are known to be involved in the infection process. PRINCIPAL FINDINGS: In the present report, we used coimmunoprecipitation and yeast two-hybrid assays to elucidate and/or confirm all the interactions that occur among the WSSV structural (envelope and tegument proteins VP51A, VP19, VP24, VP26 and VP28. We found that VP51A interacted directly not only with VP26 but also with VP19 and VP24. VP51A, VP19 and VP24 were also shown to have an affinity for self-interaction. Chemical cross-linking assays showed that these three self-interacting proteins could occur as dimers. CONCLUSIONS: From our present results in conjunction with other previously established interactions we construct a 3D model in which VP24 acts as a core protein that directly associates with VP26, VP28, VP38A, VP51A and WSV010 to form a membrane-associated protein complex. VP19 and VP37 are attached to this complex via association with VP51A and VP28, respectively. Through the VP26-VP51C interaction this envelope complex is anchored to the nucleocapsid, which is made of layers of rings formed by VP664. A 3D model of the nucleocapsid and the surrounding outer membrane is presented.

Diagnostic accuracy of spot urine protein-to-creatinine ratio for proteinuria and its association with adverse pregnancy outcomes in Chinese pregnant patients with pre-eclampsia.

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Cheung, H C; Leung, K Y; Choi, C H

2016-06-01

International guidelines have endorsed spot urine protein-to-creatinine ratio of >30 mg protein/mmol creatinine as an alternative to a 24-hour urine sample to represent significant proteinuria. This study aimed to determine the accuracy of spot urine protein-to-creatinine ratio in predicting significant proteinuria and adverse pregnancy outcome. This case series was conducted in a regional obstetric unit in Hong Kong. A total of 120 Chinese pregnant patients with pre-eclampsia delivered at Queen Elizabeth Hospital from January 2011 to December 2013 were included. Relationship of spot urine protein-to-creatinine ratio and 24-hour proteinuria; accuracy of the ratio against 24-hour urine protein at different cut-offs; and relationship of such ratio and adverse pregnancy outcome were studied. Spot urine protein-to-creatinine ratio was correlated with 24-hour urine protein with Pearson correlation coefficient of 0.914 (Pcreatinine ratio for diagnosing proteinuria in Chinese pregnant patients (33 mg/mmol) was similar to that stated in the international literature (30 mg/mmol). A cut-off of 20 mg/mmol provided a 100% sensitivity, and 52 mg/mmol provided a 100% specificity. There was no significant difference in spot urine protein-to-creatinine ratio between cases with and without adverse pregnancy outcome. Spot urine protein-to-creatinine ratio had a positive and significant correlation with 24-hour urine results in Chinese pre-eclamptic women when the ratio was <200 mg/mmol. Nonetheless, this ratio was not predictive of adverse pregnancy outcome.

Production of polyclonal antiserum specific to the 27.5 kDa envelope protein of white spot syndrome virus

NARCIS (Netherlands)

You, Z.O.; Nadala, E.C.B.; Yang, J.S.; Hulten, van M.C.W.; Loh, P.C.

2002-01-01

A truncated version of the white spot syndrome virus (WSSV) 27.5 kDa envelope protein was expressed as a histidine tag fusion protein in Escherichia coli. The bacterial expression system allowed the production of up to 10 mg of purified recombinant protein per liter of bacterial culture. Antiserum

External Quality Control for Dried Blood Spot Based C-reactive Protein Assay: Experience from the Indonesia Family Life Survey and the Longitudinal Aging Study in India

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Hu, Peifeng; Herningtyas, Elizabeth H.; Kale, Varsha; Crimmins, Eileen M.; Risbud, Arun R.; McCreath, Heather; Lee, Jinkook; Strauss, John; O’Brien, Jennifer C.; Bloom, David E.; Seeman, Teresa E.

2015-01-01

Measurement of C-reactive protein, a marker of inflammation, in dried blood spots has been increasingly incorporated in community-based social surveys internationally. Although the dried blood spot based CRP assay protocol has been validated in the United States, it remains unclear whether laboratories in other less developed countries can generate C-reactive protein results of similar quality. We therefore conducted external quality monitoring for dried blood spot based C-reactive protein measurement for the Indonesia Family Life Survey and the Longitudinal Aging Study in India. Our results show that dried blood spot based C-reactive protein results in these two countries have excellent and consistent correlations with serum-based values and dried blood spot based results from the reference laboratory in the United States. Even though the results from duplicate samples may have fluctuations in absolute values over time, the relative order of C-reactive protein levels remains similar and the estimates are reasonably precise for population-based studies that investigate the association between socioeconomic factors and health. PMID:25879265

MAMP (microbe-associated molecular pattern)-induced changes in plasma membrane-associated proteins.

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Uhlíková, Hana; Solanský, Martin; Hrdinová, Vendula; Šedo, Ondrej; Kašparovský, Tomáš; Hejátko, Jan; Lochman, Jan

2017-03-01

Plant plasma membrane associated proteins play significant roles in Microbe-Associated Molecular Pattern (MAMP) mediated defence responses including signal transduction, membrane transport or energetic metabolism. To elucidate the dynamics of proteins associated with plasma membrane in response to cryptogein, a well-known MAMP of defence reaction secreted by the oomycete Phytophthora cryptogea, 2D-Blue Native/SDS gel electrophoresis of plasma membrane fractions was employed. This approach revealed 21 up- or down-regulated protein spots of which 15 were successfully identified as proteins related to transport through plasma membrane, vesicle trafficking, and metabolic enzymes including cytosolic NADP-malic enzyme and glutamine synthetase. Observed changes in proteins were also confirmed on transcriptional level by qRT-PCR analysis. In addition, a significantly decreased accumulation of transcripts observed after employment of a mutant variant of cryptogein Leu41Phe, exhibiting a conspicuous defect in induction of resistance, sustains the contribution of identified proteins in cryptogein-triggered cellular responses. Our data provide further evidence for dynamic MAMP-induced changes in plasma membrane associated proteins. Copyright © 2016 Elsevier GmbH. All rights reserved.

Protein-induced changes during the maturation process of human dendritic cells: A 2-D DIGE approach

DEFF Research Database (Denmark)

Ferreira, Gb; Overbergh, L; Hansen, Kasper Lage

2008-01-01

Dendritic cells (DCs) are unique antigen presenting cells, which upon maturation change from a specialized antigen-capturing cell towards a professional antigen presenting cells. In this study, a 2-D DIGE analysis of immature and mature DCs was performed, to identify proteins changing in expression...... upon maturation. The protein expression profile of immature and mature DCs, derived from CD14+ peripheral blood monocytes was investigated using two pH ranges (pH 4-7 and 6-9) (n = 4). Ninety one differentially expressed spots (p...

Analysis of Tomato spotted wilt virus NSs protein indicates the importance of the N-terminal for avirulence and RNA silencing suppression

NARCIS (Netherlands)

Ronde, de D.; Pasquier, A.; Ying, S.; Butterbach, P.B.E.; Lohuis, D.; Kormelink, R.J.M.

2014-01-01

Recently, Tomato spotted wilt virus (TSWV) nonstructural protein NSs has been identified unambiguously as an avirulence (Avr) determinant for Tomato spotted wilt (Tsw)-based resistance. The observation that NSs from two natural resistance-breaking isolates had lost RNA silencing suppressor (RSS)

Proteome approaches to characterize seed storage proteins related to ditelocentric chromosomes in common wheat (Triticum aestivum L.).

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Islam, Nazrul; Woo, Sun-Hee; Tsujimoto, Hisashi; Kawasaki, Hiroshi; Hirano, Hisashi

2002-09-01

Changes in protein composition of wheat endosperm proteome were investigated in 39 ditelocentric chromosome lines of common wheat (Triticum aestivum L.) cv. Chinese Spring. Two-dimensional gel electrophoresis followed by Coomassie Brilliant Blue staining has resolved a total of 105 protein spots in a gel. Quantitative image analysis of protein spots was performed by PDQuest. Variations in protein spots between the euploid and the 39 ditelocentric lines were evaluated by spot number, appearance, disappearance and intensity. A specific spot present in all gels was taken as an internal standard, and the intensity of all other spots was calculated as the ratio of the internal standard. Out of the 1755 major spots detected in 39 ditelocentric lines, 1372 (78%) spots were found variable in different spot parameters: 147 (11%) disappeared, 978 (71%) up-regulated and 247 (18%) down-regulated. Correlation studies in changes in protein intensities among 24 protein spots across the ditelocentric lines were performed. High correlations in changes of protein intensities were observed among the proteins encoded by genes located in the homoeologous arms. Locations of structural genes controlling 26 spots were identified in 10 chromosomal arms. Multiple regulators of the same protein located at various chromosomal arms were also noticed. Identification of structural genes for most of the proteins was found difficult due to multiple regulators encoding the same protein. Two novel subunits (1B(Z,) 1BDz), the structure of which are very similar to the high molecular weight glutenin subunit 12, were identified, and the chromosome arm locations of these subunits were assigned.

Mean protein evolutionary distance: a method for comparative protein evolution and its application.

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Michael J Wise

Full Text Available Proteins are under tight evolutionary constraints, so if a protein changes it can only do so in ways that do not compromise its function. In addition, the proteins in an organism evolve at different rates. Leveraging the history of patristic distance methods, a new method for analysing comparative protein evolution, called Mean Protein Evolutionary Distance (MeaPED, measures differential resistance to evolutionary pressure across viral proteomes and is thereby able to point to the proteins' roles. Different species' proteomes can also be compared because the results, consistent across virus subtypes, concisely reflect the very different lifestyles of the viruses. The MeaPED method is here applied to influenza A virus, hepatitis C virus, human immunodeficiency virus (HIV, dengue virus, rotavirus A, polyomavirus BK and measles, which span the positive and negative single-stranded, doubled-stranded and reverse transcribing RNA viruses, and double-stranded DNA viruses. From this analysis, host interaction proteins including hemagglutinin (influenza, and viroporins agnoprotein (polyomavirus, p7 (hepatitis C and VPU (HIV emerge as evolutionary hot-spots. By contrast, RNA-directed RNA polymerase proteins including L (measles, PB1/PB2 (influenza and VP1 (rotavirus, and internal serine proteases such as NS3 (dengue and hepatitis C virus emerge as evolutionary cold-spots. The hot spot influenza hemagglutinin protein is contrasted with the related cold spot H protein from measles. It is proposed that evolutionary cold-spot proteins can become significant targets for second-line anti-viral therapeutics, in cases where front-line vaccines are not available or have become ineffective due to mutations in the hot-spot, generally more antigenically exposed proteins. The MeaPED package is available from www.pam1.bcs.uwa.edu.au/~michaelw/ftp/src/meaped.tar.gz.

Mean protein evolutionary distance: a method for comparative protein evolution and its application.

Science.gov (United States)

Wise, Michael J

2013-01-01

Proteins are under tight evolutionary constraints, so if a protein changes it can only do so in ways that do not compromise its function. In addition, the proteins in an organism evolve at different rates. Leveraging the history of patristic distance methods, a new method for analysing comparative protein evolution, called Mean Protein Evolutionary Distance (MeaPED), measures differential resistance to evolutionary pressure across viral proteomes and is thereby able to point to the proteins' roles. Different species' proteomes can also be compared because the results, consistent across virus subtypes, concisely reflect the very different lifestyles of the viruses. The MeaPED method is here applied to influenza A virus, hepatitis C virus, human immunodeficiency virus (HIV), dengue virus, rotavirus A, polyomavirus BK and measles, which span the positive and negative single-stranded, doubled-stranded and reverse transcribing RNA viruses, and double-stranded DNA viruses. From this analysis, host interaction proteins including hemagglutinin (influenza), and viroporins agnoprotein (polyomavirus), p7 (hepatitis C) and VPU (HIV) emerge as evolutionary hot-spots. By contrast, RNA-directed RNA polymerase proteins including L (measles), PB1/PB2 (influenza) and VP1 (rotavirus), and internal serine proteases such as NS3 (dengue and hepatitis C virus) emerge as evolutionary cold-spots. The hot spot influenza hemagglutinin protein is contrasted with the related cold spot H protein from measles. It is proposed that evolutionary cold-spot proteins can become significant targets for second-line anti-viral therapeutics, in cases where front-line vaccines are not available or have become ineffective due to mutations in the hot-spot, generally more antigenically exposed proteins. The MeaPED package is available from www.pam1.bcs.uwa.edu.au/~michaelw/ftp/src/meaped.tar.gz.

Time-dependent changes in protein expression in rainbow trout muscle following hypoxia.

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Wulff, Tune; Jokumsen, Alfred; Højrup, Peter; Jessen, Flemming

2012-04-18

Adaptation to hypoxia is a complex process, and individual proteins will be up- or down-regulated in order to address the main challenges at any given time. To investigate the dynamics of the adaptation, rainbow trout (Oncorhynchus mykiss) was exposed to 30% of normal oxygen tension for 1, 2, 5 and 24 h respectively, after which muscle samples were taken. The successful investigation of numerous proteins in a single study was achieved by selectively separating the sarcoplasmic proteins using 2-DE. In total 46 protein spots were identified as changing in abundance in response to hypoxia using one-way ANOVA and multivariate data analysis. Proteins of interest were subsequently identified by MS/MS following tryptic digestion. The observed regulation following hypoxia in skeletal muscle was determined to be time specific, as only a limited number of proteins were regulated in response to more than one time point. The cellular response to hypoxia included regulation of proteins involved in maintaining iron homeostasis, energy levels and muscle structure. In conclusion, this proteome-based study presents a comprehensive investigation of the expression profiles of numerous proteins at four different time points. This increases our understanding of timed changes in protein expression in rainbow trout muscle following hypoxia. Copyright © 2012 Elsevier B.V. All rights reserved.

Screening of Potential Inhibitor against Coat Protein of Apple Chlorotic Leaf Spot Virus.

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Purohit, Rituraj; Kumar, Sachin; Hallan, Vipin

2018-06-01

In this study, we analyzed Coat protein (CP) of Apple chlorotic leaf spot virus (ACLSV), an important latent virus on Apple. Incidence of the virus is upto 60% in various apple cultivars, affecting yield losses of the order of 10-40% (depending upon the cultivar). CP plays an important role as the sole building block of the viral capsid. Homology approach was used to model 193 amino acid sequence of the coat protein. We used various servers such as ConSurf, TargetS, OSML, COACH, COFACTOR for the prediction of active site residues in coat protein. Virtual screening strategy was employed to search potential inhibitors for CP. Top twenty screened molecules considered for drugability, and toxicity analysis and one potential molecule was further analyzed by docking analysis. Here, we reported a potent molecule which could inhibit the formation of viron assembly by targeting the CP protein of virus.

Rigorous assessment and integration of the sequence and structure based features to predict hot spots

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Wang Yong

2011-07-01

Full Text Available Abstract Background Systematic mutagenesis studies have shown that only a few interface residues termed hot spots contribute significantly to the binding free energy of protein-protein interactions. Therefore, hot spots prediction becomes increasingly important for well understanding the essence of proteins interactions and helping narrow down the search space for drug design. Currently many computational methods have been developed by proposing different features. However comparative assessment of these features and furthermore effective and accurate methods are still in pressing need. Results In this study, we first comprehensively collect the features to discriminate hot spots and non-hot spots and analyze their distributions. We find that hot spots have lower relASA and larger relative change in ASA, suggesting hot spots tend to be protected from bulk solvent. In addition, hot spots have more contacts including hydrogen bonds, salt bridges, and atomic contacts, which favor complexes formation. Interestingly, we find that conservation score and sequence entropy are not significantly different between hot spots and non-hot spots in Ab+ dataset (all complexes. While in Ab- dataset (antigen-antibody complexes are excluded, there are significant differences in two features between hot pots and non-hot spots. Secondly, we explore the predictive ability for each feature and the combinations of features by support vector machines (SVMs. The results indicate that sequence-based feature outperforms other combinations of features with reasonable accuracy, with a precision of 0.69, a recall of 0.68, an F1 score of 0.68, and an AUC of 0.68 on independent test set. Compared with other machine learning methods and two energy-based approaches, our approach achieves the best performance. Moreover, we demonstrate the applicability of our method to predict hot spots of two protein complexes. Conclusion Experimental results show that support vector machine

Rigorous assessment and integration of the sequence and structure based features to predict hot spots

Science.gov (United States)

2011-01-01

Background Systematic mutagenesis studies have shown that only a few interface residues termed hot spots contribute significantly to the binding free energy of protein-protein interactions. Therefore, hot spots prediction becomes increasingly important for well understanding the essence of proteins interactions and helping narrow down the search space for drug design. Currently many computational methods have been developed by proposing different features. However comparative assessment of these features and furthermore effective and accurate methods are still in pressing need. Results In this study, we first comprehensively collect the features to discriminate hot spots and non-hot spots and analyze their distributions. We find that hot spots have lower relASA and larger relative change in ASA, suggesting hot spots tend to be protected from bulk solvent. In addition, hot spots have more contacts including hydrogen bonds, salt bridges, and atomic contacts, which favor complexes formation. Interestingly, we find that conservation score and sequence entropy are not significantly different between hot spots and non-hot spots in Ab+ dataset (all complexes). While in Ab- dataset (antigen-antibody complexes are excluded), there are significant differences in two features between hot pots and non-hot spots. Secondly, we explore the predictive ability for each feature and the combinations of features by support vector machines (SVMs). The results indicate that sequence-based feature outperforms other combinations of features with reasonable accuracy, with a precision of 0.69, a recall of 0.68, an F1 score of 0.68, and an AUC of 0.68 on independent test set. Compared with other machine learning methods and two energy-based approaches, our approach achieves the best performance. Moreover, we demonstrate the applicability of our method to predict hot spots of two protein complexes. Conclusion Experimental results show that support vector machine classifiers are quite

Serum depletion induces changes in protein expression in the trophoblast-derived cell line HTR-8/SVneo.

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Novoa-Herran, Susana; Umaña-Perez, Adriana; Canals, Francesc; Sanchez-Gomez, Myriam

2016-01-01

How nutrition and growth factor restriction due to serum depletion affect trophoblast function remains poorly understood. We performed a proteomic differential study of the effects of serum depletion on a first trimester human immortalized trophoblast cell line. The viability of HTR-8/SVneo trophoblast cells in culture with 0, 0.5 and 10 % fetal bovine serum (FBS) were assayed via MTT at 24, 48 and 64 h. A comparative proteomic analysis of the cells grown with those FBS levels for 24 h was performed using two-dimensional electrophoresis (2DE), followed by mass spectrometry for protein spot identification, and a database search and bioinformatics analysis of the expressed proteins. Differential spots were identified using the Kolmogorov-Smirnov test ( n  = 3, significance level 0.10, D > 0.642) and/or ANOVA ( n  = 3, p  depletion differentially affect cell growth and protein expression. Differential expression was seen in 25 % of the protein spots grown with 0.5 % FBS and in 84 % of those grown with 0 % FBS, using 10 % serum as the physiological control. In 0.5 % FBS, this difference was related with biological processes typically affected by the serum, such as cell cycle, regulation of apoptosis and proliferation. In addition to these changes, in the serum-depleted proteome we observed downregulation of keratin 8, and upregulation of vimentin, the glycolytic enzymes enolase and pyruvate kinase (PKM2) and tumor progression-related inosine-5'-monophosphate dehydrogenase 2 (IMPDH2) enzyme. The proteins regulated by total serum depletion, but not affected by growth in 0.5 % serum, are members of the glycolytic and nucleotide metabolic pathways and the epithelial-to-mesenchymal transition (EMT), suggesting an adaptive switch characteristic of malignant cells. This comparative proteomic analysis and the identified proteins are the first evidence of a protein expression response to serum depletion in a trophoblast cell model. Our results show that

SU-E-T-510: Interplay Between Spots Sizes, Spot / Line Spacing and Motion in Spot Scanning Proton Therapy

International Nuclear Information System (INIS)

Lee, TK

2015-01-01

Purpose In proton beam configuration for spot scanning proton therapy (SSPT), one can define the spacing between spots and lines of scanning as a ratio of given spot size. If the spacing increases, the number of spots decreases which can potentially decrease scan time, and so can whole treatment time, and vice versa. However, if the spacing is too large, the uniformity of scanned field decreases. Also, the field uniformity can be affected by motion during SSPT beam delivery. In the present study, the interplay between spot/ line spacing and motion is investigated. Methods We used four Gaussian-shape spot sizes with 0.5cm, 1.0cm, 1.5cm, and 2.0cm FWHM, three spot/line spacing that creates uniform field profile which are 1/3*FWHM, σ/3*FWHM and 2/3*FWHM, and three random motion amplitudes within, +/−0.3mm, +/−0.5mm, and +/−1.0mm. We planned with 2Gy uniform single layer of 10×10cm2 and 20×20cm2 fields. Then, mean dose within 80% area of given field size, contrubuting MU per each spot assuming 1cGy/MU calibration for all spot sizes, number of spots and uniformity were calculated. Results The plans with spot/line spacing equal to or smaller than 2/3*FWHM without motion create ∼100% uniformity. However, it was found that the uniformity decreases with increased spacing, and it is more pronounced with smaller spot sizes, but is not affected by scanned field sizes. Conclusion It was found that the motion during proton beam delivery can alter the dose uniformity and the amount of alteration changes with spot size which changes with energy and spot/line spacing. Currently, robust evaluation in TPS (e.g. Eclipse system) performs range uncertainty evaluation using isocenter shift and CT calibration error. Based on presented study, it is recommended to add interplay effect evaluation to robust evaluation process. For future study, the additional interplay between the energy layers and motion is expected to present volumetric effect

Protein A from orange-spotted nervous necrosis virus triggers type I interferon production in fish cell.

Science.gov (United States)

Huang, Runqing; Zhou, Qiong; Shi, Yan; Zhang, Jing; He, Jianguo; Xie, Junfeng

2018-05-04

Family Nodaviridae consists of two genera: Alphanodavirus and Betanodavirus, and the latter is classified into four genotypes, including red-spotted grouper nervous necrosis virus, tiger puffer nervous necrosis virus, striped jack nervous necrosis virus, and barfin flounder nervous necrosis virus. Type I interferons (IFNs) play a central role in the innate immune system and antiviral responses, and the interactions between IFN and NNV have been investigated in this study. We have found that the RNA-dependent RNA polymerase (RdRp) from orange-spotted nervous necrosis virus (OGNNV), named protein A, was capable of activating IFN promoter in fathead minnow (FHM) cells. Transient expression of protein A was found to induce IFN expression and secretion, endowing FHM cells with anti-tiger frog virus ability. Protein A from SJNNV can also induce IFN expression in FHM cells but that from Flock House virus (FHV), a well-studied representative species of genus Alphanodavirus, cannot. RdRp activity and mitochondrial localization were shown to be required for protein A to induce IFN expression by means of activating IRF3 but not NFκB. Furthermore, DsRNA synthesized in vitro transcription and poly I:C activated IFN promoter activity when transfected into FHM cells, and dsRNA were also detected in NNV-infected cells. We postulated that dsRNA, a PAMP, was produced by protein A, leading to activation of innate immune response. These results suggest that protein As from NNV are the agonists of innate immune response. This is the first work to demonstrate the interaction between NNV protein A and innate immune system, and may help to understand pathogenesis of NNV. Copyright © 2018. Published by Elsevier Ltd.

Clinical utility of spot urine protein-to-creatinine ratio modified by estimated daily creatinine excretion in children.

Science.gov (United States)

Yang, Eun Mi; Yoon, Bo Ae; Kim, Soo Wan; Kim, Chan Jong

2017-06-01

The spot urine protein-to-creatinine ratio (UPCR) is widely used to predict 24-h urine protein (24-h UP) excretion. In patients with low daily urine creatinine excretion (UCr), however, the UPCR may overestimate 24-h UP. The aim of this study was to predict 24-h UP using UPCR adjusted by estimated 24-h UCr in children. This study included 442 children whose 24-h UP and spot UPCR were measured concomitantly. Estimated 24-h UCr was calculated using three previously existing equations. We estimated the 24-h UP excretion from UPCR by multiplying the estimated UCr. The results were compared with the measured 24-h UP. There was a strong correlation between UPCR and 24-h UP (r = 0.801, P < 0.001), and the correlation improved after multiplying the UPCR by the measured UCr (r = 0.847, P < 0.001). Using the estimated UCr rather than the measured UCr, there was high accuracy and strong correlation between the estimated UPCR weighted by the Cockcroft-Gault equation and 24-h UP. Improvement was also observed in the subgroup (proteinuria vs. non-proteinuria) analysis, particularly in the proteinuria group. The spot UPCR multiplied by the estimated UCr improved the accuracy of prediction of the 24-h UP in children.

Mating changes the female dietary preference in the two-spotted cricket, Gryllus bimaculatus

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Yusuke eTsukamoto

2014-03-01

Full Text Available Most insect species exhibit characteristic behavioral changes after mating. Typical post-mating behaviors in female insects include noticeable increases in food intake, elevated oviposition rates, lowered receptivity to courting males, and enhanced immune response. Although it has been reported that mated females of several insect species including the fruit fly, Drosophila melanogaster increase the amount of food intake and change their dietary preferences, the limited number of comparative studies prevent the formulation of generalities regarding post-mating behaviors in other insects in particular amongst orthopteran species. Here, we investigated whether females of the two-spotted cricket, Gryllus bimaculatus, alter their feeding behavior after mating. Although significant differences in the amount of food intake after mating were not observed, all experimental data indicated a clear trend among crickets towards the ingestion of larger quantities of food. Geometric framework analyses revealed that the mated female crickets preferred food with higher protein content compared to virgin female crickets. This implies that this species required different nutritional demands after mating. These findings further expand our understanding of the behavioral and biological changes that are triggered in female insects post-mating, and highlight the potential for this species in investigating the molecular-based nutritional dependent activities that are linked to post-mating behaviors.

Comparison of Spot Urine Protein to Creatinine Ratio to 24-Hour Proteinuria to Identify Important Change Over Time in Proteinuria in Lupus.

Science.gov (United States)

Medina-Rosas, Jorge; Su, Jiandong; Cook, Richard J; Sabapathy, Arthy; Touma, Zahi

2017-09-01

The aim of this study was to determine whether spot urine protein-to-creatinine ratio (PCR) accurately measures the change in proteinuria compared with 24-hour proteinuria (24H-P). This was a retrospective analysis on patients' paired visits and paired urine samples for PCR and 24H-P. Patients with both abnormal 24H-P (>0.5 g/d) and PCR (>0.05 g/mmol) or both normal 24H-P (≤0.5 g/d) and PCR (≤0.05 g/mmol) at baseline visit were identified.The first follow-up visit with partial recovery (50% decrease in proteinuria) or complete recovery (≤0.5 g/d) was identified for those with abnormal baseline 24H-P, and new proteinuria (>0.5 g/d) was identified for those with normal 24H-P. Twenty-four-hour urine collection and PCR end-point frequencies were compared. Twenty-four-hour urine collection results were converted to 24H-PCR. Twenty-four-hour PCR and PCR were utilized to measure the magnitude of change (by standardized response mean [SRM]) in patients who achieved the end points. Of 230 patients, at baseline, 95 patients had abnormal and 109 had normal 24H-P and PCR. On follow-up, 57 achieved partial recovery, and 53 achieved complete recovery by 24H-P. Standardized response mean was -1.03 and -1.10 for 24H-PCR and PCR, respectively. By PCR, 53 patients had partial recovery, and 27 had complete recovery. Standardized response mean was -1.25 and -0.86 by 24H-PCR and PCR, respectively.For new proteinuria, 28 patients were identified by 24H-P and 21 by PCR. Twenty-four-hour PCR SRM was 0.80, and PCR SRM was 0.68. Protein-to-creatinine ratio does not have sufficient accuracy compared with 24H-P for improvement and worsening to be used in lieu of 24H-P.

HotRegion: a database of predicted hot spot clusters.

Science.gov (United States)

Cukuroglu, Engin; Gursoy, Attila; Keskin, Ozlem

2012-01-01

Hot spots are energetically important residues at protein interfaces and they are not randomly distributed across the interface but rather clustered. These clustered hot spots form hot regions. Hot regions are important for the stability of protein complexes, as well as providing specificity to binding sites. We propose a database called HotRegion, which provides the hot region information of the interfaces by using predicted hot spot residues, and structural properties of these interface residues such as pair potentials of interface residues, accessible surface area (ASA) and relative ASA values of interface residues of both monomer and complex forms of proteins. Also, the 3D visualization of the interface and interactions among hot spot residues are provided. HotRegion is accessible at http://prism.ccbb.ku.edu.tr/hotregion.

Interleukin-1beta induced changes in the protein expression of rat islets: a computerized database

DEFF Research Database (Denmark)

Andersen, H U; Fey, S J; Larsen, Peter Mose

1997-01-01

as well as the intracellular mechanisms of action of interleukin 1-mediated beta-cell cytotoxicity are unknown. However, previous studies have found an association of beta-cell destruction with alterations in protein synthesis. Thus, two-dimensional (2-D) gel electrophoresis of pancreatic islet proteins...... may be an important tool facilitating studies of the molecular pathogenesis of insulin-dependent diabetes mellitus. 2-D gel electrophoresis of islet proteins may lead to (i) the determination of qualitative and quantitative changes in specific islet proteins induced by cytokines, (ii......) the determination of the effects of agents modulating cytokine action, and (iii) the identification of primary islet protein antigen(s) initiating the immune destruction of the beta-cells. Therefore, the aim of this study was to create databases (DB) of all reproducibly detectable protein spots on 10% and 15...

A new quantitative evaluation method for age-related changes of individual pigmented spots in facial skin.

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Kikuchi, K; Masuda, Y; Yamashita, T; Sato, K; Katagiri, C; Hirao, T; Mizokami, Y; Yaguchi, H

2016-08-01

Facial skin pigmentation is one of the most prominent visible features of skin aging and often affects perception of health and beauty. To date, facial pigmentation has been evaluated using various image analysis methods developed for the cosmetic and esthetic fields. However, existing methods cannot provide precise information on pigmented spots, such as variations in size, color shade, and distribution pattern. The purpose of this study is the development of image evaluation methods to analyze individual pigmented spots and acquire detailed information on their age-related changes. To characterize the individual pigmented spots within a cheek image, we established a simple object-counting algorithm. First, we captured cheek images using an original imaging system equipped with an illumination unit and a high-resolution digital camera. The acquired images were converted into melanin concentration images using compensation formulae. Next, the melanin images were converted into binary images. The binary images were then subjected to noise reduction. Finally, we calculated parameters such as the melanin concentration, quantity, and size of individual pigmented spots using a connected-components labeling algorithm, which assigns a unique label to each separate group of connected pixels. The cheek image analysis was evaluated on 643 female Japanese subjects. We confirmed that the proposed method was sufficiently sensitive to measure the melanin concentration, and the numbers and sizes of individual pigmented spots through manual evaluation of the cheek images. The image analysis results for the 643 Japanese women indicated clear relationships between age and the changes in the pigmented spots. We developed a new quantitative evaluation method for individual pigmented spots in facial skin. This method facilitates the analysis of the characteristics of various pigmented facial spots and is directly applicable to the fields of dermatology, pharmacology, and esthetic

Protein changes in the albedo of citrus fruits on postharvesting storage.

Science.gov (United States)

Lliso, Ignacio; Tadeo, Francisco R; Phinney, Brett S; Wilkerson, Curtis G; Talón, Manuel

2007-10-31

In this work, major protein changes in the albedo of the fruit peel of Murcott tangor (tangerine x sweet orange) during postharvest ageing were studied through 2D PAGE. Protein content in matured on-tree fruits and in fruits stored in nonstressing [99% relative humidity (RH) and 25 degrees C], cold (99% RH and 4 degrees C), and drought (60% RH and 25 degrees C) conditions was initially determined. Protein identification through MS/MS determinations revealed in all samples analyzed the occurrence of manganese superoxide dismutase (Mn SOD), actin, ATP synthase beta subunit (ATPase), citrus salt-stress associated protein (CitSap), ascorbate peroxidase (APX), translationally controlled tumor protein (TCTP), and a cysteine proteinase (CP) of the papain family. The latter protein was identified in two different gel spots, with different molecular mass, suggesting the simultaneous presence of the proteinase precursor and its active form. While Mn SOD, actin, ATPase, and CitSap were unchanged in the assayed conditions, TCTP and APX were downregulated during the postharvest ageing process. Ageing-induced APX repression was also reversed by drought. CP contents in albedo, which were similar in on- and off-tree fruits, were strongly dependent upon cold storage. The active/total CP protein ratio significantly increased after cold exposure. This proteomic survey indicates that major changes in protein content in the albedo of the peel of postharvest stored citrus fruits are apparently related to the activation of programmed cell death (PCD).

Characterization and interactome study of white spot syndrome virus envelope protein VP11.

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Wang-Jing Liu

Full Text Available White spot syndrome virus (WSSV is a large enveloped virus. The WSSV viral particle consists of three structural layers that surround its core DNA: an outer envelope, a tegument and a nucleocapsid. Here we characterize the WSSV structural protein VP11 (WSSV394, GenBank accession number AF440570, and use an interactome approach to analyze the possible associations between this protein and an array of other WSSV and host proteins. Temporal transcription analysis showed that vp11 is an early gene. Western blot hybridization of the intact viral particles and fractionation of the viral components, and immunoelectron microscopy showed that VP11 is an envelope protein. Membrane topology software predicted VP11 to be a type of transmembrane protein with a highly hydrophobic transmembrane domain at its N-terminal. Based on an immunofluorescence assay performed on VP11-transfected Sf9 cells and a trypsin digestion analysis of the virion, we conclude that, contrary to topology software prediction, the C-terminal of this protein is in fact inside the virion. Yeast two-hybrid screening combined with co-immunoprecipitation assays found that VP11 directly interacted with at least 12 other WSSV structural proteins as well as itself. An oligomerization assay further showed that VP11 could form dimers. VP11 is also the first reported WSSV structural protein to interact with the major nucleocapsid protein VP664.

Hot spot analysis for driving the development of hits into leads in fragment based drug discovery

Science.gov (United States)

Hall, David R.; Ngan, Chi Ho; Zerbe, Brandon S.; Kozakov, Dima; Vajda, Sandor

2011-01-01

Fragment based drug design (FBDD) starts with finding fragment-sized compounds that are highly ligand efficient and can serve as a core moiety for developing high affinity leads. Although the core-bound structure of a protein facilitates the construction of leads, effective design is far from straightforward. We show that protein mapping, a computational method developed to find binding hot spots and implemented as the FTMap server, provides information that complements the fragment screening results and can drive the evolution of core fragments into larger leads with a minimal loss or, in some cases, even a gain in ligand efficiency. The method places small molecular probes, the size of organic solvents, on a dense grid around the protein, and identifies the hot spots as consensus clusters formed by clusters of several probes. The hot spots are ranked based on the number of probe clusters, which predicts the binding propensity of the subsites and hence their importance for drug design. Accordingly, with a single exception the main hot spot identified by FTMap binds the core compound found by fragment screening. The most useful information is provided by the neighboring secondary hot spots, indicating the regions where the core can be extended to increase its affinity. To quantify this information, we calculate the density of probes from mapping, which describes the binding propensity at each point, and show that the change in the correlation between a ligand position and the probe density upon extending or repositioning the core moiety predicts the expected change in ligand efficiency. PMID:22145575

Fatty acid-binding protein genes of the ancient, air-breathing, ray-finned fish, spotted gar (Lepisosteus oculatus).

Science.gov (United States)

Venkatachalam, Ananda B; Fontenot, Quenton; Farrara, Allyse; Wright, Jonathan M

2018-03-01

With the advent of high-throughput DNA sequencing technology, the genomic sequence of many disparate species has led to the relatively new discipline of genomics, the study of genome structure, function and evolution. Much work has been focused on the role of whole genome duplications (WGD) in the architecture of extant vertebrate genomes, particularly those of teleost fishes which underwent a WGD early in the teleost radiation >230 million years ago (mya). Our past work has focused on the fate of duplicated copies of a multigene family coding for the intracellular lipid-binding protein (iLBP) genes in the teleost fishes. To define the evolutionary processes that determined the fate of duplicated genes and generated the structure of extant fish genomes, however, requires comparative genomic analysis with a fish lineage that diverged before the teleost WGD, such as the spotted gar (Lepisosteus oculatus), an ancient, air-breathing, ray-finned fish. Here, we describe the genomic organization, chromosomal location and tissue-specific expression of a subfamily of the iLBP genes that code for fatty acid-binding proteins (Fabps) in spotted gar. Based on this work, we have defined the minimum suite of fabp genes prior to their duplication in the teleost lineages ~230-400 mya. Spotted gar, therefore, serves as an appropriate outgroup, or ancestral/ancient fish, that did not undergo the teleost-specific WGD. As such, analyses of the spatio-temporal regulation of spotted gar genes provides a foundation to determine whether the duplicated fabp genes have been retained in teleost genomes owing to either sub- or neofunctionalization. Copyright © 2017 Elsevier Inc. All rights reserved.

Influence of Acute High Glucose on Protein Abundance Changes in Murine Glomerular Mesangial Cells

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Michelle T. Barati

2016-01-01

Full Text Available The effects of acute exposure to high glucose levels as experienced by glomerular mesangial cells in postprandial conditions and states such as in prediabetes were investigated using proteomic methods. Two-dimensional gel electrophoresis and matrix assisted laser desorption ionization time of flight mass spectrometry methods were used to identify protein expression patterns in immortalized rat mesangial cells altered by 2 h high glucose (HG growth conditions as compared to isoosmotic/normal glucose control (NG⁎ conditions. Unique protein expression changes at 2 h HG treatment were measured for 51 protein spots. These proteins could be broadly grouped into two categories: (1 proteins involved in cell survival/cell signaling and (2 proteins involved in stress response. Immunoblot experiments for a protein belonging to both categories, prohibitin (PHB, supported a trend for increased total expression as well as significant increases in an acidic PHB isoform. Additional studies confirmed the regulation of proteasomal subunit alpha-type 2 and the endoplasmic reticulum chaperone and oxidoreductase PDI (protein disulfide isomerase, suggesting altered ER protein folding capacity and proteasomal function in response to acute HG. We conclude that short term high glucose induces subtle changes in protein abundances suggesting posttranslational modifications and regulation of pathways involved in proteostasis.

Yeast ribosomal proteins

International Nuclear Information System (INIS)

Otaka, E.; Kobata, K.

1978-01-01

The cytoplasmic 80s ribosomal proteins from the cells of yeast Saccharomyces cerevisiae were analyzed by SDS two-dimensional polyacrylamide gel electrophoresis. Seventyfour proteins were identified and consecutively numbered from 1 to 74. Upon oxidation of the 80s proteins with performic acid, ten proteins (no. 15, 20, 35, 40, 44, 46, 49, 51, 54 and 55) were dislocated on the gel without change of the total number of protein spots. Five proteins (no. 8, 14, 16, 36 and 74) were phosphorylated in vivo as seen in 32 P-labelling experiments. The large and small subunits separated in low magnesium medium were analyzed by the above gel electrophoresis. At least forty-five and twenty-eight proteins were assumed to be in the large and small subunits, respectively. All proteins found in the 80s ribosomes, except for no. 3, were detected in either subunit without appearance of new spots. The acidic protein no. 3 seems to be lost during subunit dissociation. (orig.) [de

Expression, purification and crystallization of two major envelope proteins from white spot syndrome virus

International Nuclear Information System (INIS)

Tang, Xuhua; Hew, Choy Leong

2007-01-01

The crystallization of the N-terminal transmembrane region-truncated VP26 and VP28 of white spot syndrome virus is described. White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 Å resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 Å. SeMet-labelled VP28 crystallizes in space group P2 1 2 1 2 1 , with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 Å, and diffracts to 2.0 Å resolution

Expression, purification and crystallization of two major envelope proteins from white spot syndrome virus

Energy Technology Data Exchange (ETDEWEB)

Tang, Xuhua; Hew, Choy Leong, E-mail: dbshewcl@nus.edu.sg [Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543 (Singapore)

2007-07-01

The crystallization of the N-terminal transmembrane region-truncated VP26 and VP28 of white spot syndrome virus is described. White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 Å resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 Å. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 Å, and diffracts to 2.0 Å resolution.

White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp

NARCIS (Netherlands)

Hulten, van M.C.W.; Witteveldt, J.; Snippe, M.; Vlak, J.M.

2001-01-01

White spot syndrome virus (WSSV) is a large DNA virus infecting shrimp and other crustaceans. The virus particles contain at least five major virion proteins, of which three (VP26, VP24, and VP15) are present in the rod-shaped nucleocapsid and two (VP28 and VP19) reside in the envelope. The mode of

Monitoring the effects of toxic chemicals on protein expression

International Nuclear Information System (INIS)

Giometti, C.S.; Taylor, J.

1987-01-01

Two-dimensional gel electrophoresis coupled with computer-assisted image and data analysis was used to monitor protein populations for both qualitative and quantitative changes induced by exposure to chemicals. For mutagenesis studies designed to screen for heritable mutations, a computer-assisted search of the optical density data from 2DE patterns was used to look for (a) new protein spots, (b) missing protein spots and/or (c) altered expression of normal protein spots. Using this approach, 320 mice were screened for mutations induced by treatment of sires with 150 mg/kg body weight of ethylnitrosourea (ENU) and four different mutations were identified. Protein patterns from 105 offspring from untreated male mice (controls) and 369 offspring from irradiated male mice (3 Gy gamma) were also screened. No heritable mutations were found in those data sets, however. In addition, protein changes were observed in livers of animals exposed to the hepatocellular peroxisomal proliferation agents (and carcinogens) Wy-14,643 and DEHP. The de novo synthesis of a new protein by these agents was demonstrated and quantitated

Dependency on de novo protein synthesis and proteomic changes during metamorphosis of the marine bryozoan Bugula neritina

KAUST Repository

Wong, Yue Him

2010-05-24

Background: Metamorphosis in the bryozoan Bugula neritina (Linne) includes an initial phase of rapid morphological rearrangement followed by a gradual phase of morphogenesis. We hypothesized that the first phase may be independent of de novo synthesis of proteins and, instead, involves post-translational modifications of existing proteins, providing a simple mechanism to quickly initiate metamorphosis. To test our hypothesis, we challenged B. neritina larvae with transcription and translation inhibitors. Furthermore, we employed 2D gel electrophoresis to characterize changes in the phosphoproteome and proteome during early metamorphosis. Differentially expressed proteins were identified by liquid chromatography tandem mass spectrometry and their gene expression patterns were profiled using semi-quantitative real time PCR.Results: When larvae were incubated with transcription and translation inhibitors, metamorphosis initiated through the first phase but did not complete. We found a significant down-regulation of 60 protein spots and the percentage of phosphoprotein spots decreased from 15% in the larval stage to12% during early metamorphosis. Two proteins--the mitochondrial processing peptidase beta subunit and severin--were abundantly expressed and phosphorylated in the larval stage, but down-regulated during metamorphosis. MPPbeta and severin were also down-regulated on the gene expression level.Conclusions: The initial morphogenetic changes that led to attachment of B. neritina did not depend on de novo protein synthesis, but the subsequent gradual morphogenesis did. This is the first time that the mitochondrial processing peptidase beta subunit or severin have been shown to be down-regulated on both gene and protein expression levels during the metamorphosis of B. neritina. Future studies employing immunohistochemistry to reveal the expression locality of these two proteins during metamorphosis should provide further evidence of the involvement of these two

Antiviral RNA silencing suppression activity of Tomato spotted wilt virus NSs protein.

Science.gov (United States)

Ocampo Ocampo, T; Gabriel Peralta, S M; Bacheller, N; Uiterwaal, S; Knapp, A; Hennen, A; Ochoa-Martinez, D L; Garcia-Ruiz, H

2016-06-17

In addition to regulating gene expression, RNA silencing is an essential antiviral defense system in plants. Triggered by double-stranded RNA, silencing results in degradation or translational repression of target transcripts. Viruses are inducers and targets of RNA silencing. To condition susceptibility, most plant viruses encode silencing suppressors that interfere with this process, such as the Tomato spotted wilt virus (TSWV) NSs protein. The mechanism by which NSs suppresses RNA silencing and its role in viral infection and movement remain to be determined. We cloned NSs from the Hawaii isolate of TSWV and using two independent assays show for the first time that this protein restored pathogenicity and supported the formation of local infection foci by suppressor-deficient Turnip mosaic virus and Turnip crinkle virus. Demonstrating the suppression of RNA silencing directed against heterologous viruses establishes the foundation to determine the means used by NSs to block this antiviral process.

Managing emerging threats to spotted owls

Science.gov (United States)

Ho Yi Wan; Joseph L. Ganey; Christina D. Vojta; Samuel A. Cushman

2018-01-01

The 3 spotted owl (Strix occidentalis) subspecies in North America (i.e., northern spotted owl [S. o. caurina], California spotted owl [S. o. occidentalis], Mexican spotted owl [S. o. lucida]) have all experienced population declines over the past century due to habitat loss and fragmentation from logging. Now, the emerging influences of climate change, high-severity...

Envelope Proteins of White Spot Syndrome Virus (WSSV Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT.

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Shijun Xie

Full Text Available White spot syndrome virus (WSSV is a major pathogen in shrimp cultures. The interactions between viral proteins and their receptors on the surface of cells in a frontier target tissue are crucial for triggering an infection. In this study, a yeast two-hybrid (Y2H library was constructed using cDNA obtained from the stomach and gut of Litopenaeus vannamei, to ascertain the role of envelope proteins in WSSV infection. For this purpose, VP37 was used as the bait in the Y2H library screening. Forty positive clones were detected after screening. The positive clones were analyzed and discriminated, and two clones belonging to the peritrophin family were subsequently confirmed as genuine positive clones. Sequence analysis revealed that both clones could be considered as the same gene, LV-peritrophin (LvPT. Co-immunoprecipitation confirmed the interaction between LvPT and VP37. Further studies in the Y2H system revealed that LvPT could also interact with other WSSV envelope proteins such as VP32, VP38A, VP39B, and VP41A. The distribution of LvPT in tissues revealed that LvPT was mainly expressed in the stomach than in other tissues. In addition, LvPT was found to be a secretory protein, and its chitin-binding ability was also confirmed.

Plasma immune protein analysis in the orange-spotted grouper Epinephelus coioides: Evidence for altered expressions of immune factors associated with a choline-supplemented diet.

Science.gov (United States)

Shiu, Ya-Li; Chiu, Kuo-Hsun; Huynh, Truong-Giang; Liu, Ping-Chung; Liu, Chun-Hung

2017-06-01

This study aimed to unravel the regulatory roles of choline in activating immune responses and disease resistance of the orange-spotted grouper Epinephelus coioides. Fish were fed a choline-supplemented diet at 1 g kg -1 of feed for 30 days. Fish fed a fish meal basal diet without choline-supplement served as controls. At the end of the feeding trial, fish were challenged with Vibrio alginolyticus. Meanwhile, plasma proteomics of fish in each group were also evaluated by two-dimensional gel electrophoresis (2-DE), and differentially expressed proteins were identified by tandem mass spectrophotometry (MS/MS), then a Western blot analysis or real-time polymerase chain reaction was used to confirm differential expressions of immune-enhancing proteins. Results showed that choline significantly increased survival of E. coioides 48 days after being injected with V. alginolyticus. From maps of plasma proteins, a comparative analysis between the control and choline groups revealed that 111 spots matched, with 26 altered expression spots in the choline group. Of these 26 spots, 16 were upregulated and 10 downregulated. After protein identification by reverse-phase nano-high-performance liquid chromatography-electrospray ionization MS/MS analysis, eight of 26 proteins were found to be immune-related proteins, all of which were upregulated, including complement 3 (C3), alpha-2-macroglobulin-P-like isoform (A2M), fibrinogen beta chain precursor (FBG), and immunoglobulin heavy constant mu (Ighm) proteins. Expression of the A2M protein and A2M enzyme activity in plasma of fish fed choline significantly increased compared to the control group. Additionally, A2M messenger (m)RNA transcripts were also upregulated in the liver and kidneys. Significantly higher C3 expressions at both the mRNA and protein levels were detected in the liver of fish in the choline group. Moreover, FBG gene expressions in the liver and kidneys significantly increased, while Ighm increased in the

Detection of ligand binding hot spots on protein surfaces via fragment-based methods: application to DJ-1 and glucocerebrosidase

Energy Technology Data Exchange (ETDEWEB)

Landon, Melissa R.; Lieberman, Raquel L.; Hoang, Quyen Q.; Ju, Shulin; Caaveiro, Jose M.M.; Orwig, Susan D.; Kozakov, Dima; Brenke, Ryan; Chuang, Gwo-Yu; Beglov, Dmitry; Vajda, Sandor; Petsko, Gregory A.; Ringe, Dagmar; (BU-M); (Brandeis); (GIT)

2010-08-04

The identification of hot spots, i.e., binding regions that contribute substantially to the free energy of ligand binding, is a critical step for structure-based drug design. Here we present the application of two fragment-based methods to the detection of hot spots for DJ-1 and glucocerebrosidase (GCase), targets for the development of therapeutics for Parkinson's and Gaucher's diseases, respectively. While the structures of these two proteins are known, binding information is lacking. In this study we employ the experimental multiple solvent crystal structures (MSCS) method and computational fragment mapping (FTMap) to identify regions suitable for the development of pharmacological chaperones for DJ-1 and GCase. Comparison of data derived via MSCS and FTMap also shows that FTMap, a computational method for the identification of fragment binding hot spots, is an accurate and robust alternative to the performance of expensive and difficult crystallographic experiments.

Microbiological and biochemical changes in pearl spot (Etroplus suratensis Bloch) stored under modified atmospheres.

Science.gov (United States)

Lalitha, K V; Sonaji, E R; Manju, S; Jose, L; Gopal, T K S; Ravisankar, C N

2005-01-01

This study aimed to determine the effect of packaging [air, modified atmosphere (MA)] on microbial growth, sensory and chemical parameters and also on shelf life of fresh pearl spot (Etroplus suratensis Bloch) and on the selection of microbial association. Fresh pearl spot (whole, gutted) were packaged under both 100% air and MAs (40%CO(2)/60% O(2), 50%CO(2)/50%O(2), 60% CO(2)/40%O(2), 70% CO(2)/30% O(2) and 40% CO(2)/30% O(2)/30% N(2)) and stored at 0 degrees C. Microbial growth (counts of total aerobic bacteria, H(2)S-producing bacteria, Lactic acid bacteria, Brochothrix thermosphacta, yeast and mould), chemical spoilage indicators (pH, total volatile basic nitrogen) and sensory characteristics were monitored. Microbial changes in Pearl spot packed under 100% air and 40% CO(2)/30%O(2)/30% N(2) were similar. The total volatile basic nitrogen values increased, but the values never exceeded the acceptability limit of 25 mg 100 g(-1). MA 60% CO(2) : 40%O(2) was found to be better with a shelf life of 21 days whereas air stored samples had a shelf-life of 12-14 days only. Storage of pearl spot under MAs 60% CO(2) : 40%O(2) is a promising method to extend shelf-life. Longer shelf life expands the market potential of pearl spot and reduces waste during distribution and retail display.

Comparative proteomic analysis of oil palm leaves infected with Ganoderma boninense revealed changes in proteins involved in photosynthesis, carbohydrate metabolism, and immunity and defense.

Science.gov (United States)

Jeffery Daim, Leona Daniela; Ooi, Tony Eng Keong; Ithnin, Nalisha; Mohd Yusof, Hirzun; Kulaveerasingam, Harikrishna; Abdul Majid, Nazia; Karsani, Saiful Anuar

2015-08-01

The basidiomycete fungal pathogen Ganoderma boninense is the causative agent for the incurable basal stem rot (BSR) disease in oil palm. This disease causes significant annual crop losses in the oil palm industry. Currently, there is no effective method for disease control and elimination, nor is any molecular marker for early detection of the disease available. An understanding of how BSR affects protein expression in plants may help identify and/or assist in the development of an early detection protocol. Although the mode of infection of BSR disease is primarily via the root system, defense-related genes have been shown to be expressed in both the root and leafs. Thus, to provide an insight into the changes in the global protein expression profile in infected plants, comparative 2DE was performed on leaf tissues sampled from palms with and without artificial inoculation of the Ganoderma fungus. Comparative 2DE revealed that 54 protein spots changed in abundance. A total of 51 protein spots were successfully identified by LC-QTOF MS/MS. The majority of these proteins were those involved in photosynthesis, carbohydrate metabolism as well as immunity and defense. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Advances in spot curing technology

International Nuclear Information System (INIS)

Burga, R.

1999-01-01

A brief review of spot curing technology was presented. The process which a spot of energy of a specific wavelength bandwidth and irradiance is used to cause a coating, encapsulant or adhesive to change from a liquid to a solid state

Height Resolution of Antibody Spots Measured by Spinning-Disk Interferometry on the BioCD

Directory of Open Access Journals (Sweden)

Kevin O’Brien

2016-02-01

Full Text Available Spinning-disc interferometry (SDI is a high-speed laser scanning approach to surface metrology that uses common-path interferometry to measure protein spots on a BioCD disk. The measurement sensitivity depends on the scanning pitch and on the time-base. Based on high-resolution laser scanning images of printed antibody spots, we quantify the protein sensitivity as a function of the scan parameters. For smoothly printed antibody spots scanned with a transverse spatial resolution of 1 μm, the surface height precision for a single 100 μm diameter protein spot is approximately 1 pm. This detection sensitivity sets the fundamental limit of detection for label-free BioCD biosensors performing immunoassays.

Investigation of Hardness Change for Spot Welded Tailored Blank in Hot Stamping Using CCT and Deformation-CCT Diagrams

Science.gov (United States)

Yogo, Yasuhiro; Kurato, Nozomi; Iwata, Noritoshi

2018-04-01

When an outer panel of a B-pillar is manufactured with the hot stamping process, reinforcements are spot welded on its inner side. Before reinforcements are added, the microstructure of the outer panel is martensite. However, reheating during spot welding changes the martensite to ferrite, which has a lower hardness in the heat-affected zone than in other areas. If spot welding is conducted before hot stamping for making a spot welded tailored blank, the microstructure in the spot welded tailored blank after hot stamping is martensite. This sequence of processes avoids hardness reduction due to spot welding. In this study, the hardness and microstructure around spot welded parts of the tailored blank were investigated. The results clearly showed that areas close to the spot welded parts are severely stretched during hot stamping. In addition, stretching suppresses the martensitic phase transformation and reduces the hardness. To characterize this phenomenon, a simulation was conducted that considered the effects of pre-strain on the phase transformation. A continuous cooling transformation (CCT) diagram and a deformation continuous cooling transformation (DCCT) diagram were made in order to quantify the effect of the cooling rate and pre-strain on the phase transformation and hardness. The hardness was then calculated using the experimentally measured CCT and DCCT diagrams and the finite element analysis results. The calculated hardness was compared with the experimental hardness. Good agreement was found between the calculated and experimental results.

Investigation of Hardness Change for Spot Welded Tailored Blank in Hot Stamping Using CCT and Deformation-CCT Diagrams

Science.gov (United States)

Yogo, Yasuhiro; Kurato, Nozomi; Iwata, Noritoshi

2018-06-01

When an outer panel of a B-pillar is manufactured with the hot stamping process, reinforcements are spot welded on its inner side. Before reinforcements are added, the microstructure of the outer panel is martensite. However, reheating during spot welding changes the martensite to ferrite, which has a lower hardness in the heat-affected zone than in other areas. If spot welding is conducted before hot stamping for making a spot welded tailored blank, the microstructure in the spot welded tailored blank after hot stamping is martensite. This sequence of processes avoids hardness reduction due to spot welding. In this study, the hardness and microstructure around spot welded parts of the tailored blank were investigated. The results clearly showed that areas close to the spot welded parts are severely stretched during hot stamping. In addition, stretching suppresses the martensitic phase transformation and reduces the hardness. To characterize this phenomenon, a simulation was conducted that considered the effects of pre-strain on the phase transformation. A continuous cooling transformation (CCT) diagram and a deformation continuous cooling transformation (DCCT) diagram were made in order to quantify the effect of the cooling rate and pre-strain on the phase transformation and hardness. The hardness was then calculated using the experimentally measured CCT and DCCT diagrams and the finite element analysis results. The calculated hardness was compared with the experimental hardness. Good agreement was found between the calculated and experimental results.

Changes of serum contents of LPO, SOD after treatment with vita. E-C complex in patients with liver spot

International Nuclear Information System (INIS)

Li Qing; Feng Zheng

2005-01-01

Objective: To investigate the changes of serum contents of LPO, SOD and therapeutic efficacy after treatment with Vita. E-C complex in patients with liver spot. Methods: Serum LPO and SOD contents were measured both before and after treatment with Vita E-C complex (Vita. E l00mg, Vita. C 200mg x 3/d for 3 months) in 30 patients with liver spot as well as in 10 controls. Results: Before treatment, the serum LPO contents in the patients were significantly higher than those in controls. After treatment, the LPO contents dropped markedly, being significantly lower than the values before treatment. However, the SOD contents were about the same as those in controls and changes little after treatment. Conclusion: Vita. E-C complex was of definite therapeutic value for the treatment of liver spot. (authors)

A single point mutation in Tomato spotted wilt virus NSs protein is sufficient to overcome Tsw-gene-mediated resistance in pepper.

Science.gov (United States)

Almási, Asztéria; Nemes, Katalin; Csömör, Zsófia; Tóbiás, István; Palkovics, László; Salánki, Katalin

2017-06-01

The nonstructural protein (NSs) of Tomato spotted wilt virus (TSWV) was previously identified as an avirulence determinant for Tsw-based resistance on pepper. The NSs of wild-type (WT) and resistance-breaking (RB) TSWV strains isolated in Hungary had only two amino acid substitutions (104, 461). We have analysed the ability of the NSs and their point mutant variants to trigger Tsw-mediated hypersensitive responses and RNA silencing suppressor (RSS) activity in patch assays. We identified a single amino acid change at position 104 (T-A) that was responsible for the necrosis induction or loss, while a significant difference was not detected in the RSS activity of the two parental strains. We have successfully complemented the infection of the WT strain on resistant pepper cultivar with the infectious S RNA transcript of the RB strain and the WT-T104A point mutant. Our work provides direct evidence that a single amino acid change can induce an RB phenotype.

Analysis of Tomato spotted wilt virus NSs protein indicates the importance of the N-terminal domain for avirulence and RNA silencing suppression.

Science.gov (United States)

de Ronde, Dryas; Pasquier, Adrien; Ying, Su; Butterbach, Patrick; Lohuis, Dick; Kormelink, Richard

2014-02-01

Recently, Tomato spotted wilt virus (TSWV) nonstructural protein NSs has been identified unambiguously as an avirulence (Avr) determinant for Tomato spotted wilt (Tsw)-based resistance. The observation that NSs from two natural resistance-breaking isolates had lost RNA silencing suppressor (RSS) activity and Avr suggested a link between the two functions. To test this, a large set of NSs mutants was generated by alanine substitutions in NSs from resistance-inducing wild-type strains (NSs(RI) ), amino acid reversions in NSs from resistance-breaking strains (NSs(RB)), domain deletions and swapping. Testing these mutants for their ability to suppress green fluorescent protein (GFP) silencing and to trigger a Tsw-mediated hypersensitive response (HR) revealed that the two functions can be separated. Changes in the N-terminal domain were found to be detrimental for both activities and indicated the importance of this domain, additionally supported by domain swapping between NSs(RI) and NSs(RB). Swapping domains between the closely related Tospovirus Groundnut ringspot virus (GRSV) NSs and TSWV NSs(RI) showed that Avr functionality could not simply be transferred between species. Although deletion of the C-terminal domain rendered NSs completely dysfunctional, only a few single-amino-acid mutations in the C-terminus affected both functions. Mutation of a GW/WG motif (position 17/18) rendered NSs completely dysfunctional for RSS and Avr activity, and indicated a putative interaction between NSs and Argonaute 1 (AGO1), and its importance in TSWV virulence and viral counter defence against RNA interference. © 2013 BSPP AND JOHN WILEY & SONS LTD.

Dynamic changes in proteins during apple (Malus x domestica) fruit ripening and storage

OpenAIRE

Shi, Yun; Jiang, Li; Zhang, Li; Kang, Ruoyi; Yu, Zhifang

2014-01-01

A proteomic study, using two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight, was conducted in apple fruit (cv. ‘Golden Delicious’) starting at 10 days prior to harvest through 50 days in storage. Total protein was extracted using a phenol/sodium dodecyl sulfate protocol. More than 400 protein spots were detected in each gel and 55 differentially expressed proteins (p

Estimasi Sintesis Protein Mikrobia Rumen Menggunakan Ekskresi Derivat Purin dalam Urin dengan Teknik Spot Sampling pada Kambing Bligon dan Kambing Kejobong

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Dianestu Putra

2016-11-01

Full Text Available This study were aimed to determine the correlation between concentration of purine derivatives (PD in spot sample with PD total excretion in Bligon and Kejobong goats and determine the appropriate sampling time, in order to predicting microbial protein synthesis in both breeds. Six male Bligon goats and six male Kejobong goats with age range from 8 to 14 months and body weight from 16 to 21 kg were placed in metabolism cages. Peanut straw and water were given to both groups of goats through ad libitum feeding and drinking. The study was done in 14 days for adaptation, 3 days for collection. Sample of feeds, feed residues, and feces were collected and then analyzed to determine dry matter and organic matter content. Spot urine and the total daily urine samples were also collected. Samples collection of spot sampling technique was run by taking the urine periodically with 3 hours intervals at 24 hours. Urine samples were analyzed for the content of creatinine and PD which includes allantoin, uric acid, xanthine, and hypoxanthine. Data were tested for the correlation between concentration of PD spot urine sample with total PD daily excretion. It is known that the concentration of PD and creatinine (µmol/L for Bligon were 1,418.40 and 202.85 respectively, while for Kejobong were 1,547.40 and 219.68 respectively. Total excretion of PD, allantoin, uric acid, xanthyne and hypoxanthine and creatinine (µmol/W0,75/day for Bligon were 114.14, 95.86, 17.31, 0.97, and 16.40 respectively, with microbial protein synthesis efficiency was 4.61 g N/kg degraded of organic matter in rumen (DOMR. Total excretion of PD allantoin, uric acid, xanthyne and hypoxanthine and creatinine (µmol/W0,75/day for Kejobong were 180.18, 158.17, 20.60, 1.40, and 24.87 respectively, with microbial protein synthesis efficiency was 6.90 g N/kg DOMR. Based on this study also known that the best time for spot sampling to determine the total excretion of PD in Bligon was in the range

Suitability of two-dimensional electrophoretic protein separations for quantitative detection of mutations

International Nuclear Information System (INIS)

Taylor, J.; Anderson, N.L.; Anderson, N.G.; Gemmell, A.; Giometti, C.S.; Nance, S.L.; Tollaksen, S.L.

1986-01-01

Separation of proteins by two-dimensional electrophoresis (2DE) provides a powerful method for mutagenesis studies, since hundreds of proteins can be monitored simultaneously. In previous mutation studies in which 2DE has been used, only qualitative protein differences were monitored; quantitative protein variations were not evaluated. Although significant differences in protein abundance can be detected by eye, the large number of protein spots present in 2DE patterns together with the large number of individual patterns required for a mutagenesis study would necessitate the use of a computerized analysis system to detect the rare quantitative protein changes indicative of gene deletions or inactivation of genes by point mutations in regulatory genes. A pilot study to search for heritable mutations induced by treatment of mice with either ethylnitrosourea or gamma radiation is underway. Samples are being monitored for quantitative changes that reduce the amount of protein by about 50%. The results of this study indicate that the key methods to improve the application of 2DE to mutation screening are to increase the number of measurable spots (i.e., improve stain sensitivity) and to decrease the spread of values for the volume measurements. Even small improvements in these areas could greatly increase the number of monitorable spots. 9 refs., 4 figs

Spot Urine Protein-to-Creatinine Ratio to Predict the Magnitude of 24-Hour Total Proteinuria in Preeclampsia of Varying Severity.

Science.gov (United States)

Kucukgoz Gulec, Umran; Sucu, Mete; Ozgunen, Fatma Tuncay; Buyukkurt, Selim; Guzel, Ahmet Baris; Paydas, Saime

2017-10-01

The predictive value of spot urine protein-to-creatinine ratio (PCR) for estimating total 24-hour proteinuria in severe preeclampsia is unclear. This study aimed to assess the diagnostic accuracy of spot urine PCR for ascertaining the magnitude of proteinuria in women with preeclampsia of varying severity. A total of 205 patients with prediagnosed preeclampsia were included in this prospective cohort study. Patients were allocated into one of the three groups categorized by severity of disease, as follows: gestational hypertension, group 1 (n = 41); preeclampsia, group 2 (n = 88); and severe preeclampsia, group 3 (n = 76). We assessed the spot urine PCRs to determine significant proteinuria and the magnitude of proteinuria in these groups. The spot urine PCR was 0.53, with 81% sensitivity and 93% specificity to detect significant proteinuria. A significant correlation was found between PCR and 24-hour total proteinuria in group 1 (r = 0.473, P = 0.002). There were also significant correlations in group 2 (r = 0.814, P spot urine PCR to estimate 24-hour total proteinuria in severe preeclampsia was Y = 832.02X + 378.74 mg (r 2  = 0.8304). Although 24-hour urine collection remains a merely reliable test to determine the degree of total proteinuria, our findings suggest that it is likely to assess the magnitude of proteinuria by the spot urine PCR, especially in severe preeclampsia. www.clinicaltrials.govNCT01623791. Copyright © 2017 The Society of Obstetricians and Gynaecologists of Canada/La Société des obstétriciens et gynécologues du Canada. Published by Elsevier Inc. All rights reserved.

Proteomic Analysis of Copper-Binding Proteins in Excess Copper-Stressed Roots of Two Rice (Oryza sativa L. Varieties with Different Cu Tolerances.

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Chen Chen

Full Text Available To better understand the mechanisms involved in the heavy metal stress response and tolerance in plants, a proteomic approach was used to investigate the differences in Cu-binding protein expression in Cu-tolerant and Cu-sensitive rice varieties. Cu-binding proteins from Cu-treated rice roots were separated using a new IMAC method in which an IDA-sepharose column was applied prior to the Cu-IMAC column to remove metal ions from protein samples. More than 300 protein spots were reproducibly detected in the 2D gel. Thirty-five protein spots exhibited changes greater than 1.5-fold in intensity compared to the control. Twenty-four proteins contained one or more of nine putative metal-binding motifs reported by Smith et al., and 19 proteins (spots contained one to three of the top six motifs reported by Kung et al. The intensities of seven protein spots were increased in the Cu-tolerant variety B1139 compared to the Cu-sensitive variety B1195 (p<0.05 and six protein spots were markedly up-regulated in B1139, but not detectable in B1195. Four protein spots were significantly up-regulated in B1139, but unchanged in B1195 under Cu stress. In contrast, two protein spots were significantly down-regulated in B1195, but unchanged in B1139. These Cu-responsive proteins included those involved in antioxidant defense and detoxification (spots 5, 16, 21, 22, 28, 29 and 33, pathogenesis (spots 5, 16, 21, 22, 28, 29 and 33, regulation of gene transcription (spots 8 and 34, amino acid synthesis (spots 8 and 34, protein synthesis, modification, transport and degradation (spots 1, 2, 4, 10, 15, 19, 30, 31, 32 and 35, cell wall synthesis (spot 14, molecular signaling (spot 3, and salt stress (spots 7, 9 and 27; together with other proteins, such as a putative glyoxylate induced protein, proteins containing dimeric alpha-beta barrel domains, and adenosine kinase-like proteins. Our results suggest that these proteins, together with related physiological processes, play

Hand-Held Photometer for Instant On-Spot Quantification of Nucleic Acids, Proteins, and Cells.

Science.gov (United States)

Li, Shi-Hao; Jain, Abhinav; Tscharntke, Timo; Arnold, Tobias; Trau, Dieter W

2018-02-20

This paper presents a novel hand-held photometer, termed "Photopette", for on-spot absorbance measurements of biochemical analytes. The Photopette is a multicomponent, highly portable device with an overall weight of 160 g, which fits within 202 mm × 47 mm × 42 mm. Designed in the form factor of a micropipette, Photopette integrates a photodiode detector with light emitting diodes (LEDs) to form a highly customizable photometer which supports a wide variety of applications within the wavelengths between 260 and 1050 nm. A dual-purpose disposable reflective tip was designed to act as a sample holder and a light-reflecting system, which is in stark contrast to the operation of mainstream spectrophotometers and photometers. Small volume analytes may be measured with low sample loss using this proprietary CuveTip. A user-friendly software application running on smart devices was developed to control and read the values from Photopette via a low-energy Bluetooth link. This one-step strategy allows measurements on-spot without sample transfer, minimizing cross-contamination and human error. The results reported in this paper demonstrate Photopette's great potential to quantify DNA, direct protein, and cell density directly within the laminar flow hood. Results are compared with a Nanodrop 2000c spectrophotometer, a mainstream spectrophotometer for small-volume measurements.

Lessons from hot spot analysis for fragment-based drug discovery

Science.gov (United States)

Hall, David R.; Vajda, Sandor

2015-01-01

Analysis of binding energy hot spots at protein surfaces can provide crucial insights into the prospects for successful application of fragment-based drug discovery (FBDD), and whether a fragment hit can be advanced into a high affinity, druglike ligand. The key factor is the strength of the top ranking hot spot, and how well a given fragment complements it. We show that published data are sufficient to provide a sophisticated and quantitative understanding of how hot spots derive from protein three-dimensional structure, and how their strength, number and spatial arrangement govern the potential for a surface site to bind to fragment-sized and larger ligands. This improved understanding provides important guidance for the effective application of FBDD in drug discovery. PMID:26538314

Cellular imaging by targeted assembly of hot-spot SERS and photoacoustic nanoprobes using split-fluorescent protein scaffolds.

Science.gov (United States)

Köker, Tuğba; Tang, Nathalie; Tian, Chao; Zhang, Wei; Wang, Xueding; Martel, Richard; Pinaud, Fabien

2018-02-09

The in cellulo assembly of plasmonic nanomaterials into photo-responsive probes is of great interest for many bioimaging and nanophotonic applications but remains challenging with traditional nucleic acid scaffolds-based bottom-up methods. Here, we address this quandary using split-fluorescent protein (FP) fragments as molecular glue and switchable Raman reporters to assemble gold or silver plasmonic nanoparticles (NPs) into photonic clusters directly in live cells. When targeted to diffusing surface biomarkers in cancer cells, the NPs self-assemble into surface-enhanced Raman-scattering (SERS) nanoclusters having hot spots homogenously seeded by the reconstruction of full-length FPs. Within plasmonic hot spots, autocatalytic activation of the FP chromophore and near-field amplification of its Raman fingerprints enable selective and sensitive SERS imaging of targeted cells. This FP-driven assembly of metal colloids also yields enhanced photoacoustic signals, allowing the hybrid FP/NP nanoclusters to serve as contrast agents for multimodal SERS and photoacoustic microscopy with single-cell sensitivity.

Quantitative assessment of local perfusion change in acute intracerebral hemorrhage areas with and without "dynamic spot sign" using CT perfusion imaging.

Science.gov (United States)

Fu, Fan; Sui, Binbin; Liu, Liping; Su, Yaping; Sun, Shengjun; Li, Yingying

2018-01-01

Background Positive "dynamic spot sign" has been proven to be a potential risk factor for acute intracerebral hemorrhage (ICH) expansion, but local perfusion change has not been quantitatively investigated. Purpose To quantitatively evaluate perfusion changes at the ICH area using computed tomography perfusion (CTP) imaging. Material and Methods Fifty-three patients with spontaneous ICH were recruited. Unenhanced computed tomography (NCCT), CTP within 6 h, and follow-up NCCT were performed for 21 patients in the "spot sign"-positive group and 32 patients in the control group. Cerebral perfusion change was quantitatively measured on regional cerebral blood flow/regional cerebral blood volume (rCBF/rCBV) maps. Regions of interest (ROIs) were set at the "spot-sign" region and the whole hematoma area for "spot-sign"-positive cases, and at one of the highest values of three interested areas and the whole hematoma area for the control group. Hematoma expansion was determined by follow-up NCCT. Results For the "spot-sign"-positive group, the average rCBF (rCBV) values at the "spot-sign" region and the whole hematoma area were 21.34 ± 15.24 mL/min/100 g (21.64 ± 21.48 mL/100g) and 5.78 ± 6.32 mL/min/100 g (6.07 ± 5.45 mL/100g); for the control group, the average rCBF (rCBV) values at the interested area and whole hematoma area were 2.50 ± 1.83 mL/min/100 g (3.13 ± 1.96 mL/100g) and 3.02 ± 1.80 mL/min/100 g (3.40 ± 1.44 mL/100g), respectively. Average rCBF and rCBV values of the "spot-sign" region were significantly different from other regions ( P spot-sign"-positive and control groups were 25.24 ± 19.38 mL and -0.41 ± 1.34 mL, respectively. Conclusion The higher perfusion change at ICH on CTP images may reflect the contrast extravasation and be associated with the hematoma expansion.

ENVIRONMENTAL CHANGES ANALYSIS IN BUCHAREST CITY USING CORONA, SPOT HRV AND IKONOS IMAGES

Directory of Open Access Journals (Sweden)

I. Noaje

2012-08-01

Full Text Available Bucharest, capital of Romania, deals with serious difficulties as a result of urban politics: influx of people due to industrialization and development of dormitory areas, lack of a modern infrastructure, absence of coherent and long term urban development politics, continuous depletion of environment. This paper presents a multisensor study relying on multiple data sets, both analogical and digital: satellite images (Corona – 1964 panchromatic, SPOT HRV – 1994 multispctral and panchromatic, IKONOS – 2007 multispectral, aerial photographs – 1994, complementary products (topographic and thematic maps. Georeferenced basis needs to be generated to highlight changes detection. The digital elevation model is generated from aerial photography 1:5,000 scaled, acquired in 1994. First a height correction is required followed by an affine transformation to the ground control points identified both in aerial photographs and IKONOS image. SPOT-HRV pansharpened satellite image has been rectified on georeferenced IKONOS image, by an affine transformation method. The Corona panoramic negative film was scanned and rubber sheeting method is used for rectification. The first 25 years of the study period (1964–1989 are characterized by growth of industrial areas, high density apartment buildings residential areas and leisure green areas by demolition of cultural heritage areas (hundred years old churches and architectural monuments. Changes between the imagery were determined partially through visual interpretation, using elements such as location, size, shape, shadow, tone, texture, and pattern (Corona image, partially using unsupervised classification (SPOT HRV and IKONOS. The second period of 18 years (1989–2007 highlighted considerable growth of residential areas in the city neighborhood, simultaneously with the diminish of green areas and massive deforestation in confiscated areas before and returned to the original owners.

Dramatic Change in Jupiter's Great Red Spot from Spacecraft Observations

Science.gov (United States)

Simon, Amy A.; Wong, Michael H.; Rogers, John H.; Orton, Glenn S.; de Pater, Imke; Asay-Davis, Xylar; Carlson, Robert W.; Marcus, Philip S.

2015-01-01

Jupiter's Great Red Spot (GRS) is one of its most distinct and enduring features. Since the advent of modern telescopes, keen observers have noted its appearance and documented a change in shape from very oblong to oval, confirmed in measurements from spacecraft data. It currently spans the smallest latitude and longitude size ever recorded. Here we show that this change has been accompanied by an increase in cloud/haze reflectance as sensed in methane gas absorption bands, increased absorption at wavelengths shorter than 500 nanometers, and increased spectral slope between 500 and 630 nanometers. These changes occurred between 2012 and 2014, without a significant change in internal tangential wind speeds; the decreased size results in a 3.2 day horizontal cloud circulation period, shorter than previously observed. As the GRS has narrowed in latitude, it interacts less with the jets flanking its north and south edges, perhaps allowing for less cloud mixing and longer UV irradiation of cloud and aerosol particles. Given its long life and observational record, we expect that future modeling of the GRS's changes, in concert with laboratory flow experiments, will drive our understanding of vortex evolution and stability in a confined flow field crucial for comparison with other planetary atmospheres.

Proteomic analysis of barley response during early spot blotch infection

International Nuclear Information System (INIS)

Al-Daoude, A.; Jawhar, M.; Shoaib, A.; Arabi, M.I.E.

2015-01-01

Spot blotch (SB), caused by the fungus Cochliobolus sativus, is a common foliar disease of barley worldwide, but little is known about the host response to infection at the protein level. In this study, a systematic shotgun proteomics approach was chosen to document the early barley response to C. sativus infection. Overall, 28 protein spots were consistently observed as differential in the proteome profiles of the challenged and unchallenged plants. After tryptic digestion, MALDI-TOF/MS analysis and MASCOT database searching identified proteins associated with the defense response including resistance proteins, putative hydrolase, proteinase, kinase and general metabolism and transport proteins. These afford important functions in host resistance and pathogen's inhibition in plants. One of the identified products is a putative NBS-LRR protein which is considered one of the major plant disease resistance proteins identified to date. This work indicates that, in combination with functional genomics, response of barley to challenge by C. sativus involved the recruitment of proteins from various defense pathways.(author)

Spot Urine-guided Salt Reduction in Chronic Kidney Disease Patients.

Science.gov (United States)

Uchiyama, Kiyotaka; Yanai, Akane; Ishibashi, Yoshitaka

2017-09-01

Dietary salt restriction is important in patients with chronic kidney disease (CKD) to reduce hypertension, cardiovascular events, progression of CKD, and mortality. However, recommending salt reduction for patients is difficult without knowing their actual sodium intake. This study evaluated the effectiveness of spot urine-guided salt reduction in CKD outpatients. A prospective cohort study was used. This study included a total of 127 adult outpatients (aged 60 ± 18 years, 80 males) with CKD. Their baseline estimated glomerular filtration rate was 51.4 ± 25.1 (mL/minute/1.73 m 2 ), and 64 (50%) of them were with CKD stage 3a or 3b (both 32 [25%]). We informed the patients of their individual spot urine-estimated salt intake every time they visited the outpatient clinic. Based on the data, the nephrologist encouraged the patients to achieve their salt restriction goal. The primary outcome was the estimated salt excretion, and the secondary outcome was the urinary protein-to-Cr ratio (UPCR). Multiple regression analyses were performed to clarify the contributing factors of changes in both outcomes. Over a follow-up of 12 months, the median number of patients' visits was 7 (5-8). The estimated salt intake was significantly reduced from 7.98 ± 2.49 g/day to 6.77 ± 1.77 g/day (P intake, with borderline significance (P = .08). Providing spot urine-estimated salt intake feedback effectively motivated CKD patients to reduce their salt intake. Spot urine-guided salt reduction may slow CKD progression through decreased urinary protein excretion. Copyright © 2017 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

Time Resolved X-Ray Spot Size Diagnostic

CERN Document Server

Richardson, Roger; Falabella, Steven; Guethlein, Gary; Raymond, Brett; Weir, John

2005-01-01

A diagnostic was developed for the determination of temporal history of an X-ray spot. A pair of thin (0.5 mm) slits image the x-ray spot to a fast scintillator which is coupled to a fast detector, thus sampling a slice of the X-Ray spot. Two other scintillator/detectors are used to determine the position of the spot and total forward dose. The slit signal is normalized to the dose and the resulting signal is analyzed to get the spot size. The position information is used to compensate for small changes due to spot motion and misalignment. The time resolution of the diagnostic is about 1 ns and measures spots from 0.5 mm to over 3 mm. The theory and equations used to calculate spot size and position are presented, as well as data. The calculations assume a symmetric, Gaussian spot. The spot data is generated by the ETA II accelerator, a 2kA, 5.5 MeV, 60ns electron beam focused on a Tantalum target. The spot generated is typically about 1 mm FWHM. Comparisons are made to an X-ray pinhole camera which images th...

The silencing suppressor (NSs) protein of the plant virus Tomato spotted wilt virus enhances heterologous protein expression and baculovirus pathogenicity in cells and lepidopteran insects.

Science.gov (United States)

de Oliveira, Virgínia Carla; da Silva Morgado, Fabricio; Ardisson-Araújo, Daniel Mendes Pereira; Resende, Renato Oliveira; Ribeiro, Bergmann Morais

2015-11-01

In this work, we showed that cell death induced by a recombinant (vAcNSs) Autographa californica multiple nucleopolyhedrovirus (AcMNPV) expressing the silencing suppressor (NSs) protein of Tomato spotted wilt virus (TSWV) was enhanced on permissive and semipermissive cell lines. The expression of a heterologous gene (firefly luciferase) during co-infection of insect cells with vAcNSs and a second recombinant baculovirus (vAgppolhfluc) was shown to increase when compared to single vAgppolhfluc infections. Furthermore, the vAcNSs mean time-to-death values were significantly lower than those for wild-type AcMNPV on larvae of Spodoptera frugiperda and Anticarsia gemmatalis. These results showed that the TSWV-NSs protein could efficiently increase heterologous protein expression in insect cells as well as baculovirus pathogenicity and virulence, probably by suppressing the gene-silencing machinery in insects.

Dramatic Change in Jupiter's Great Red Spot

Science.gov (United States)

Simon, A. A.; Wong, M. H.; Rogers, J. H.; Orton, G. S.; de Pater, I.; Asay-Davis, X.; Carlson, R. W.; Marcus, P. S.

2015-01-01

Jupiter's Great Red Spot (GRS) is one of its most distinct and enduring features, having been continuously observed since the 1800's. It currently spans the smallest latitude and longitude size ever recorded. Here we show analyses of 2014 Hubble spectral imaging data to study the color, structure and internal dynamics of this long-live storm.

Lessons from Hot Spot Analysis for Fragment-Based Drug Discovery.

Science.gov (United States)

Hall, David R; Kozakov, Dima; Whitty, Adrian; Vajda, Sandor

2015-11-01

Analysis of binding energy hot spots at protein surfaces can provide crucial insights into the prospects for successful application of fragment-based drug discovery (FBDD), and whether a fragment hit can be advanced into a high-affinity, drug-like ligand. The key factor is the strength of the top ranking hot spot, and how well a given fragment complements it. We show that published data are sufficient to provide a sophisticated and quantitative understanding of how hot spots derive from a protein 3D structure, and how their strength, number, and spatial arrangement govern the potential for a surface site to bind to fragment-sized and larger ligands. This improved understanding provides important guidance for the effective application of FBDD in drug discovery. Copyright © 2015 Elsevier Ltd. All rights reserved.

Changes in protein composition and protein phosphorylation during ...

African Journals Online (AJOL)

Changes in protein profiles and protein phosphorylation were studied in various stages of germinating somatic and zygotic embryos. Many proteins, which were expressed in cotyledonary stage somatic embryos, were also present in the zygotic embryos obtained from mature dry seed. The intensity of 22 kDa protein was ...

Hot spot manifestation in eclipsing dwarf nova HT Cassiopeiae

OpenAIRE

Bakowska, K.; Olech, A.

2014-01-01

We report the detection of the hot spot in light curves of the eclipsing dwarf nova HT Cassiopeiae during its superoutburst in 2010 November. Analysis of eight reconstructed light curves of the hot spot eclipses showed directly that the brightness of the hot spot was changing significantly during the superoutburst. Thereby, detected hot spot manifestation in HT Cas is the newest observational evidence for the EMT model for dwarf novae.

Accurate prediction of hot spot residues through physicochemical characteristics of amino acid sequences

KAUST Repository

Chen, Peng; Li, Jinyan; Limsoon, Wong; Kuwahara, Hiroyuki; Huang, Jianhua Z.; Gao, Xin

2013-01-01

Hot spot residues of proteins are fundamental interface residues that help proteins perform their functions. Detecting hot spots by experimental methods is costly and time-consuming. Sequential and structural information has been widely used in the computational prediction of hot spots. However, structural information is not always available. In this article, we investigated the problem of identifying hot spots using only physicochemical characteristics extracted from amino acid sequences. We first extracted 132 relatively independent physicochemical features from a set of the 544 properties in AAindex1, an amino acid index database. Each feature was utilized to train a classification model with a novel encoding schema for hot spot prediction by the IBk algorithm, an extension of the K-nearest neighbor algorithm. The combinations of the individual classifiers were explored and the classifiers that appeared frequently in the top performing combinations were selected. The hot spot predictor was built based on an ensemble of these classifiers and to work in a voting manner. Experimental results demonstrated that our method effectively exploited the feature space and allowed flexible weights of features for different queries. On the commonly used hot spot benchmark sets, our method significantly outperformed other machine learning algorithms and state-of-the-art hot spot predictors. The program is available at http://sfb.kaust.edu.sa/pages/software.aspx. © 2013 Wiley Periodicals, Inc.

Accurate prediction of hot spot residues through physicochemical characteristics of amino acid sequences

KAUST Repository

Chen, Peng

2013-07-23

Hot spot residues of proteins are fundamental interface residues that help proteins perform their functions. Detecting hot spots by experimental methods is costly and time-consuming. Sequential and structural information has been widely used in the computational prediction of hot spots. However, structural information is not always available. In this article, we investigated the problem of identifying hot spots using only physicochemical characteristics extracted from amino acid sequences. We first extracted 132 relatively independent physicochemical features from a set of the 544 properties in AAindex1, an amino acid index database. Each feature was utilized to train a classification model with a novel encoding schema for hot spot prediction by the IBk algorithm, an extension of the K-nearest neighbor algorithm. The combinations of the individual classifiers were explored and the classifiers that appeared frequently in the top performing combinations were selected. The hot spot predictor was built based on an ensemble of these classifiers and to work in a voting manner. Experimental results demonstrated that our method effectively exploited the feature space and allowed flexible weights of features for different queries. On the commonly used hot spot benchmark sets, our method significantly outperformed other machine learning algorithms and state-of-the-art hot spot predictors. The program is available at http://sfb.kaust.edu.sa/pages/software.aspx. © 2013 Wiley Periodicals, Inc.

Accurate prediction of hot spot residues through physicochemical characteristics of amino acid sequences.

Science.gov (United States)

Chen, Peng; Li, Jinyan; Wong, Limsoon; Kuwahara, Hiroyuki; Huang, Jianhua Z; Gao, Xin

2013-08-01

Hot spot residues of proteins are fundamental interface residues that help proteins perform their functions. Detecting hot spots by experimental methods is costly and time-consuming. Sequential and structural information has been widely used in the computational prediction of hot spots. However, structural information is not always available. In this article, we investigated the problem of identifying hot spots using only physicochemical characteristics extracted from amino acid sequences. We first extracted 132 relatively independent physicochemical features from a set of the 544 properties in AAindex1, an amino acid index database. Each feature was utilized to train a classification model with a novel encoding schema for hot spot prediction by the IBk algorithm, an extension of the K-nearest neighbor algorithm. The combinations of the individual classifiers were explored and the classifiers that appeared frequently in the top performing combinations were selected. The hot spot predictor was built based on an ensemble of these classifiers and to work in a voting manner. Experimental results demonstrated that our method effectively exploited the feature space and allowed flexible weights of features for different queries. On the commonly used hot spot benchmark sets, our method significantly outperformed other machine learning algorithms and state-of-the-art hot spot predictors. The program is available at http://sfb.kaust.edu.sa/pages/software.aspx. Copyright © 2013 Wiley Periodicals, Inc.

Semi-Supervised Transductive Hot Spot Predictor Working on Multiple Assumptions

KAUST Repository

Wang, Jim Jing-Yan

2014-05-23

Protein-protein interactions are critically dependent on just a few residues (“hot spots”) at the interfaces. Hot spots make a dominant contribution to the binding free energy and if mutated they can disrupt the interaction. As mutagenesis studies require significant experimental efforts, there exists a need for accurate and reliable computational hot spot prediction methods. Compared to the supervised hot spot prediction algorithms, the semi-supervised prediction methods can take into consideration both the labeled and unlabeled residues in the dataset during the prediction procedure. The transductive support vector machine has been utilized for this task and demonstrated a better prediction performance. To the best of our knowledge, however, none of the transductive semi-supervised algorithms takes all the three semisupervised assumptions, i.e., smoothness, cluster and manifold assumptions, together into account during learning. In this paper, we propose a novel semi-supervised method for hot spot residue prediction, by considering all the three semisupervised assumptions using nonlinear models. Our algorithm, IterPropMCS, works in an iterative manner. In each iteration, the algorithm first propagates the labels of the labeled residues to the unlabeled ones, along the shortest path between them on a graph, assuming that they lie on a nonlinear manifold. Then it selects the most confident residues as the labeled ones for the next iteration, according to the cluster and smoothness criteria, which is implemented by a nonlinear density estimator. Experiments on a benchmark dataset, using protein structure-based features, demonstrate that our approach is effective in predicting hot spots and compares favorably to other available methods. The results also show that our method outperforms the state-of-the-art transductive learning methods.

Neutralization of White Spot Syndrome Virus by Monoclonal Antibodies against Viral Envelope Proteins

Directory of Open Access Journals (Sweden)

Hsiu-Hui Shih

2004-09-01

Full Text Available Two monoclonal antibodies (MAbs recognizing envelope proteins of the white spot syndrome virus (WSSV, 6E1 against VP28 and 3E8 against VP19, were applied to demonstrate their neutralizing ability to this virus by using both in vitro and in vivo assays. Mixtures of MAb 6E1 with virus filtrate were inoculated into the primary explant monolayer culture derived from the lymphoid Oka organs of Penaeus monodon. Mab was likely to neutralize the infectivity of virus to monolayer since cytopathic effects were apparently blocked in experiment group. WSSV was titrated using Blue-Cell ELISA and the neutralizing index was calculated to be 6.90 for 6EI and 5.83 for 3E8. Neutralized virus fluids injected intramuscularly into post larvae of P. monodon. The shrimp in the positive control, which were injected with WSSV only showed an increasing mortality and a 100% mortality was reached at day 34, whereas no shrimp died in the negative control. The mortality for 6E1 was 6.7% and for 3E8 was 13.3%. These results suggest that Mabs recognizing the WSSV envelope proteins could neutralize viral infectivity to both cultured cells and shrimp.

SpotADAPT

DEFF Research Database (Denmark)

Kaulakiene, Dalia; Thomsen, Christian; Pedersen, Torben Bach

2015-01-01

by Amazon Web Services (AWS). The users aiming for the spot market are presented with many instance types placed in multiple datacenters in the world, and thus it is difficult to choose the optimal deployment. In this paper, we propose the framework SpotADAPT (Spot-Aware (re-)Deployment of Analytical...... of typical analytical workloads and real spot price traces. SpotADAPT's suggested deployments are comparable to the theoretically optimal ones, and in particular, it shows good cost benefits for the budget optimization -- on average SpotADAPT is at most 0.3% more expensive than the theoretically optimal...

A Sightseeing Spot Recommendation System That Takes into Account the Change in Circumstances of Users

Directory of Open Access Journals (Sweden)

Yuri Mizutani

2017-10-01

Full Text Available The present study aimed to design, develop, operate and evaluate a sightseeing spot recommendation system for urban sightseeing spots in order to support individual, as well as group sightseeing activities while taking into consideration the user’s needs, which can change according to the circumstances (each user’s important conditions and sightseeing unit. The system was developed by integrating Web-GIS (Geographic Information Systems, the pairing system, the evaluation system, as well as the recommendation system into a single system, and it was also connected with external SNS (Social Networking Services: Twitter and Facebook. Additionally, the system was operated for four weeks in the central part of Yokohama City in Kanagawa Prefecture, Japan, and the total number of users was 52. Based on the results of the web questionnaire survey, the usefulness of the system when sightseeing was high, and the recommendation function of sightseeing spots, which is an original function, has received mainly good ratings. From the results of the access analysis of users’ log data, it is evident that the system has been used by different types of devices, just as it was designed for, and that the system has been used according to the purpose of the present study, which is to support the sightseeing activities of users.

The novel white spot syndrome virus-induced gene, PmERP15, encodes an ER stress-responsive protein in black tiger shrimp, Penaeus monodon.

Science.gov (United States)

Leu, Jiann-Horng; Liu, Kuan-Fu; Chen, Kuan-Yu; Chen, Shu-Hwa; Wang, Yu-Bin; Lin, Chung-Yen; Lo, Chu-Fang

2015-04-01

By microarray screening, we identified a white spot syndrome virus (WSSV)-strongly induced novel gene in gills of Penaeus monodon. The gene, PmERP15, encodes a putative transmembrane protein of 15 kDa, which only showed some degree of similarity (54-59%) to several unknown insect proteins, but had no hits to shrimp proteins. RT-PCR showed that PmERP15 was highly expressed in the hemocytes, heart and lymphoid organs, and that WSSV-induced strong expression of PmERP15 was evident in all tissues examined. Western blot analysis likewise showed that WSSV strongly up-regulated PmERP15 protein levels. In WSSV-infected hemocytes, immunofluorescence staining showed that PmERP15 protein was colocalized with an ER enzyme, protein disulfide isomerase, and in Sf9 insect cells, PmERP15-EGFP fusion protein colocalized with ER -Tracker™ Red dye as well. GRP78, an ER stress marker, was found to be up-regulated in WSSV-infected P. monodon, and both PmERP15 and GRP78 were up-regulated in shrimp injected with ER stress inducers tunicamycin and dithiothreitol. Silencing experiments showed that although PmERP15 dsRNA-injected shrimp succumbed to WSSV infection more rapidly, the WSSV copy number had no significant changes. These results suggest that PmERP15 is an ER stress-induced, ER resident protein, and its induction in WSSV-infected shrimp is caused by the ER stress triggered by WSSV infection. Furthermore, although PmERP15 has no role in WSSV multiplication, its presence is essential for the survival of WSSV-infected shrimp. Copyright © 2014 Elsevier Ltd. All rights reserved.

Comparison of 24-hour urinary protein and protein-to-creatinine ratio in the assessment of proteinuria

International Nuclear Information System (INIS)

Wahbeh, Ayman M; Ewais, Mohammad H; Elsharif, Mahamed E

2009-01-01

To determine the correlation between protein-to-creatinine ratio (PCR) and 24-hour urinary protein (UP), we measured proteinuria in 68 patients attending the nephrology clinic at Jordan University Hospital by 24-hour urine protein excretion and protein-to-creatinine ratio. The cutoff values for spot urine protein-to-creatinine ratio in predicting 24-hour protein 'threshold' excretion of 0.5, 1.0 and 3.5 g/day were determined using receiver operating characteristic curves. A very good correlation (r= 0.832, P< 0.0001) was found between spot urine protein-to-creatinine ratio and 24-hour urine protein excretion. Bland-Altman plot showed the two tests had reasonable limits of agreement at low level of protein excretion but the limits became wider as the protein excretion increased. For protein excretion < 2.0 g/day, the limits of agreement of spot urine (PCR) and (UP) were +1.48 and -1.2 g/day. The spot urine protein-to-creatinine ratios of 0.72 (sensitivity 0.97; specificity 1.0), 1.2 (0.97; 0.89) and 3.23 (1.0; 0.86) mg/mg reliably predicted 24-hour urine total protein equivalent 'thresholds' of 0.5, 1.0 and 3.5 g/day, respectively. We conclude that the protein-to-creatinine ratio in spot urine specimens is an accurate, convenient, and reliable method to estimate the protein excretion in urine. However, the protein-to-creatinine ratio will likely be within clinically acceptable limits only when proteinuria is at reasonably low levels. (author)

Alterations of proteins in MDCK cells during acute potassium deficiency.

Science.gov (United States)

Peerapen, Paleerath; Ausakunpipat, Nardtaya; Chanchaem, Prangwalai; Thongboonkerd, Visith

2016-06-01

Chronic K(+) deficiency can cause hypokalemic nephropathy associated with metabolic alkalosis, polyuria, tubular dilatation, and tubulointerstitial injury. However, effects of acute K(+) deficiency on the kidney remained unclear. This study aimed to explore such effects by evaluating changes in levels of proteins in renal tubular cells during acute K(+) deficiency. MDCK cells were cultivated in normal K(+) (NK) (K(+)=5.3 mM), low K(+) (LK) (K(+)=2.5 mM), or K(+) depleted (KD) (K(+)=0 mM) medium for 24 h and then harvested. Cellular proteins were resolved by two-dimensional gel electrophoresis (2-DE) and visualized by SYPRO Ruby staining (5 gels per group). Spot matching and quantitative intensity analysis revealed a total 48 protein spots that had significantly differential levels among the three groups. Among these, 46 and 30 protein spots had differential levels in KD group compared to NK and LK groups, respectively. Comparison between LK and NK groups revealed only 10 protein spots that were differentially expressed. All of these differentially expressed proteins were successfully identified by Q-TOF MS and/or MS/MS analyses. The altered levels of heat shock protein 90 (HSP90), ezrin, lamin A/C, tubulin, chaperonin-containing TCP1 (CCT1), and calpain 1 were confirmed by Western blot analysis. Global protein network analysis showed three main functional networks, including 1) cell growth and proliferation, 2) cell morphology, cellular assembly and organization, and 3) protein folding in which the altered proteins were involved. Further investigations on these networks may lead to better understanding of pathogenic mechanisms of low K(+)-induced renal injury. Copyright © 2016 Elsevier B.V. All rights reserved.

White Spot Syndrome Virus infection in Penaeus monodon is ...

Indian Academy of Sciences (India)

White Spot Syndrome Virus (WSSV) is a major pathogen in shrimp aquaculture, and its rampant spread has resulted in great economic loss. Identification of host cellular proteins interacting with WSSV will help in unravelling the repertoire of host proteins involved in WSSV infection. In this study, we have employed ...

Evaluation of the use of purine derivatives:creatinine ratio in spot urine samples as an index of microbial protein supply in Yerli Kara crossbred cattle

International Nuclear Information System (INIS)

Cetinkaya, N.; Ozdemir, H.; Gucus, A.I.; Ozcan, H.; Sogut, A.; Yaman, S.

2004-01-01

In Experiment I, response of daily purine derivatives (PD) excretion to feed intake in Yerli Kara crossbred (YK-C) cattle on state farms was measured. Animals were fed a mixed diet containing 30% wheat straw and 70% compounded feed. Crude protein and organic matter contents of the diet were 12.4% and 95%, respectively. In Experiment II, spot urine sampling technique was applied at state farm. Four Yerli Kara crossbred bulls with a mean live weight of 211.0 ± 41.3 kg were used. The experimental design, feeding and diet were the same as in Experiment I. The treatments were allocated according to a 4 x 4 Latin Square design. In Experiment III, spot urine sampling technique was applied at smallholder farms. Two to three kg of compound feed (crude protein 12%) containing 65% barley, 25% bran, 6% sunflower seed meal, 3% marmer dust and 1% mineral and vitamin mixture was offered in two parts, one in the morning (0730 h) and the other in the afternoon (1700 h). The ingredients in the compound feed were similar for all animals, but animals in Groups I, II and III received 1 to 2 kg/d of straw (crude protein 3%), grass hay (crude protein 7%), or both straw and grass hay respectively. In Experiment I, a significant correlation (R 2 =0.99) between PD excretion (Y, mmol/d) and digestible organic matter intake, DOMI (X, kg/d) for YK-C cattle was observed (Y = 12.5 + 19.7 X). Moreover, daily PD excretion (mmol/d) was correlated with the PDC index, which was defined as [PD molar concentration] / [Creatinine molar concentration] x kgW 0.75 . In Experiment II, the PDC index increased with level of intake. The coefficient of variation due to time of sampling for uric acid, allantoin, PD, creatinine, total-N, the PDC Index in spot urine samples were less than 5%. In Experiment III, the PDC index were 49.95 ± 13.5, 45.6 ± 13.0, 48.95 ± 15.3 for the three groups respectively. These values were similar to those for 60% intake level in Experiment I. Using the equation DOMI = 344 + 48

Proteomic identification of rhythmic proteins in rice seedlings.

Science.gov (United States)

Hwang, Heeyoun; Cho, Man-Ho; Hahn, Bum-Soo; Lim, Hyemin; Kwon, Yong-Kook; Hahn, Tae-Ryong; Bhoo, Seong Hee

2011-04-01

Many aspects of plant metabolism that are involved in plant growth and development are influenced by light-regulated diurnal rhythms as well as endogenous clock-regulated circadian rhythms. To identify the rhythmic proteins in rice, periodically grown (12h light/12h dark cycle) seedlings were harvested for three days at six-hour intervals. Continuous dark-adapted plants were also harvested for two days. Among approximately 3000 reproducible protein spots on each gel, proteomic analysis ascertained 354 spots (~12%) as light-regulated rhythmic proteins, in which 53 spots showed prolonged rhythm under continuous dark conditions. Of these 354 ascertained rhythmic protein spots, 74 diurnal spots and 10 prolonged rhythmic spots under continuous dark were identified by MALDI-TOF MS analysis. The rhythmic proteins were functionally classified into photosynthesis, central metabolism, protein synthesis, nitrogen metabolism, stress resistance, signal transduction and unknown. Comparative analysis of our proteomic data with the public microarray database (the Plant DIURNAL Project) and RT-PCR analysis of rhythmic proteins showed differences in rhythmic expression phases between mRNA and protein, suggesting that the clock-regulated proteins in rice are modulated by not only transcriptional but also post-transcriptional, translational, and/or post-translational processes. 2011 Elsevier B.V. All rights reserved.

Glacier Changes in the Cordillera Blanca, Peru, Derived From SPOT5 Imagery, GIS and Field- Based Measurements

Science.gov (United States)

Racoviteanu, A.; Arnaud, Y.; Williams, M. W.; Singh Khalsa, S.

2007-12-01

There is urgency in deriving an extensive dataset for deriving glacier changes within the Cordillera Blanca, Peru, in a cost-effective and timely manner. Rapid glacial retreat during the last decades in this area poses a threat for water resources, hydroelectric power and local traditions. While there is some information on decadal changes in glacier extents, there still remains a paucity of mass balance measurements and glacier parameters such as hypsometry, size distribution and termini elevations. Here we investigate decadal changes in glacier parameters for Cordillera Blanca of Peru using data from Système Probatoire d'Observation de la Terre (SPOT) sensor, an old glacier inventory from 1970 aerial photography, field-based mass balance measurements and meteorological observations. Here we focus on: constructing a geospatial glacier inventory from 2003 SPOT scenes; mass balance estimations using remote sensing and field data; frequency distribution of glacier area; changes in termini elevations; hypsometry changes over time; glacier topography (slope, aspect, length/width ratio); AAR vs. mass balance for Artesonraju and Yanamarey benchmark glaciers; precipitation and temperature trends in the region. Over the last 25 years, mean temperatures increases of 0.09 deg.C/yr were greater at lower elevation than the 0.01 deg.C/yr at higher elevations, with little change in precipitation. Comparison of the new SPOT-based glacier inventory with the 1970 inventory shows that glaciers in Cordillera Blanca retreated at a rate of 0.6% per year over the last three decades, with no significant differences in the rate of area loss between E and W side. At lower elevations there is an upward shift of glacier termini along with a decrease in glacier area. Small glaciers are losing more area than large glaciers. Based on the relationship between specific mass balance (bn) and accumulation area ratio (AAR) for the two benchmark glaciers, we predicted a steady-state equilibrium line

Acute Heat Stress Changes Protein Expression in the Testes of a Broiler-Type Strain of Taiwan Country Chickens.

Science.gov (United States)

Wang, Shih-Han; Cheng, Chuen-Yu; Chen, Chao-Jung; Chan, Hong-Lin; Chen, Hsin-Hsin; Tang, Pin-Chi; Chen, Chih-Feng; Lee, Yen-Pai; Huang, San-Yuan

2018-03-19

Heat stress leads to decreased fertility in roosters. This study investigated the global protein expression in response to acute heat stress in the testes of a broiler-type strain of Taiwan country chickens (TCCs). Twelve 45-week-old roosters were randomly allocated to the control group maintained at 25°C, and three groups subjected to acute heat stress at 38°C for 4 h, with 0, 2, and 6 h of recovery, respectively. Testis samples were collected for hematoxylin and eosin staining, apoptosis assay, and protein analysis. The results revealed 101 protein spots that differed significantly from the control following exposure to acute heat stress. The proteins that were differentially expressed participated mainly in protein metabolism and other metabolic processes, responses to stimuli, apoptosis, cellular organization, and spermatogenesis. Proteins that negatively regulate apoptosis were downregulated and proteins involved in autophagy and major heat shock proteins (HSP90α, HSPA5, and HSPA8) were upregulated in the testes of heat-stressed chickens. In conclusion, acute heat stress causes a change in protein expression in the testes of broiler-type B strain TCCs and may thus impair cell morphology, spermatogenesis, and apoptosis. The expression of heat shock proteins increased to attenuate the testicular injury induced by acute heat stress.

Spot protein-creatinine ratio and spot albumin-creatinine ratio in the assessment of pre-eclampsia: a diagnostic accuracy study with decision-analytic model-based economic evaluation and acceptability analysis.

Science.gov (United States)

Waugh, Jason; Hooper, Richard; Lamb, Edmund; Robson, Stephen; Shennan, Andrew; Milne, Fiona; Price, Christopher; Thangaratinam, Shakila; Berdunov, Vladislav; Bingham, Jenn

2017-10-01

The National Institute for Health and Care Excellence (NICE) guidelines highlighted the need for 'large, high-quality prospective studies comparing the various methods of measuring proteinuria in women with new-onset hypertensive disorders during pregnancy'. The primary objective was to evaluate quantitative assessments of spot protein-creatinine ratio (SPCR) and spot albumin-creatinine ratio (SACR) in predicting severe pre-eclampsia (PE) compared with 24-hour urine protein measurement. The secondary objectives were to investigate interlaboratory assay variation, to evaluate SPCR and SACR thresholds in predicting adverse maternal and fetal outcomes and to assess the cost-effectiveness of these models. This was a prospective diagnostic accuracy cohort study, with decision-analytic modelling and a cost-effectiveness analysis. The setting was 36 obstetric units in England, UK. Pregnant women (aged ≥ 16 years), who were at > 20 weeks' gestation with confirmed gestational hypertension and trace or more proteinuria on an automated dipstick urinalysis. Women provided a spot urine sample for protein analysis (the recruitment sample) and were asked to collect a 24-hour urine sample, which was stored for secondary analysis. A further spot sample of urine was taken immediately before delivery. Outcome data were collected from hospital records. There were four index tests on a spot sample of urine: (1) SPCR test (conducted at the local laboratory); (2) SPCR test [conducted at the central laboratory using the benzethonium chloride (BZC) assay]; (3) SPCR test [conducted at the central laboratory using the pyrogallol red (PGR) assay]; and (4) SACR test (conducted at the central laboratory using an automated chemistry analyser). The comparator tests on 24-hour urine collection were a central test using the BZC assay and a central test using the PGR assay. The primary reference standard was the NICE definition of severe PE. Secondary reference standards were a clinician

Spatial cognitive deficits in an animal model of Wernicke-Korsakoff syndrome are related to changes in thalamic VDAC protein concentrations.

Science.gov (United States)

Bueno, K O; de Souza Resende, L; Ribeiro, A F; Dos Santos, D M; Gonçalves, E C; Vigil, F A B; de Oliveira Silva, I F; Ferreira, L F; de Castro Pimenta, A M; Ribeiro, A M

2015-05-21

Proteomic profiles of the thalamus and the correlation between the rats' performance on a spatial learning task and differential protein expression were assessed in the thiamine deficiency (TD) rat model of Wernicke-Korsakoff syndrome. Two-dimensional gel-electrophoresis detected 320 spots and a significant increase or decrease in seven proteins. Four proteins were correlated to rat behavioral performance in the Morris Water Maze. One of the four proteins was identified by mass spectrometry as Voltage-Dependent Anion Channels (VDACs). The association of VDAC is evident in trials in which the rats' performance was worst, in which the VDAC protein was reduced, as confirmed by Western blot. No difference was observed on the mRNA of Vdac genes, indicating that the decreased VDAC expression may be related to a post-transcriptional process. The results show that TD neurodegeneration involves changes in thalamic proteins and suggest that VDAC protein activity might play an important role in an initial stage of the spatial learning process. Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.

Variability of Jupiter's Five-Micron Hot Spot Inventory

Science.gov (United States)

Yanamandra-Fisher, Padma A.; Orton, G. S.; Wakefield, L.; Rogers, J. H.; Simon-Miller, A. A.; Boydstun, K.

2012-01-01

Global upheavals on Jupiter involve changes in the albedo of entire axisymmetric regions, lasting several years, with the last two occurring in 1989 and 2006. Against this backdrop of planetary-scale changes, discrete features such as the Great Red Spot (GRS), and other vortices exhibit changes on shorter spatial- and time-scales. We track the variability of the discrete equatorial 5-micron hot spots, semi-evenly spaced in longitude and confined to a narrow latitude band centered at 6.5degN (southern edge of the North Equatorial Belt, NEB), abundant in Voyager images. Tantalizingly similar patterns were observed in the visible (bright plumes and blue-gray regions), where reflectivity in the red is anti-correlated with 5-microns thermal radiance. Ortiz et al. (1998, GRL, 103) characterized the latitude and drift rates of the hot spots, including the descent of the Galileo probe at the southern edge of a 5-micron hot spot, as the superposition of equatorial Rossby waves, with phase speeds between 99 - 103m/s, relative to System III. We note that the high 5-micron radiances correlate well but not perfectly with high 8.57-micron radiances. Because the latter are modulated primarily by changes in the upper ammonia (NH3) ice cloud opacity, this correlation implies that changes in the ammonia ice cloud field may be responsible for the variability seen in the 5-m maps. During the NEB fade (2011 - early 2012), however, these otherwise ubiquitous features were absent, an atmospheric state not seen in decades. The ongoing NEB revival indicates nascent 5-m hot spots as early as April 2012, with corresponding visible dark spots. Their continuing growth through July 2012 indicates the possit.le re-establishment of Rossby waves. The South Equatorial Belt (SEB) and NEB revivals began similarly with an instability that developed into a major outbreak, and many similarities in the observed propagation of clear regions.

Preliminary study on plasma proteins in pregnant and non-pregnant female dogs.

Science.gov (United States)

Szczubiał, Marek; Wawrzykowski, Jacek; Dąbrowski, Roman; Krawczyk, Magdalena; Kankofer, Marta

2017-07-15

In this study, we used a combined approach based on 2-dimensional electrophoresis (2-DE), difference in gel electrophoresis (DIGE), and mass spectrometry (MS) to identify the plasma protein composition in pregnant female dogs and compared it with non-pregnant female dogs. We used the plasma samples obtained from four female dogs during I, II, and III thirds of pregnancy, three days after parturition, as well as from four non-pregnant female dogs in diestrus phase. Analysis of 2-DE gel image exhibited of 249 protein spots. The intensity of staining of 35 spots differed significantly (P dogs. We used matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI TOF MS) to identify 47 spots corresponding to 52 different proteins. Five identified protein spots, including zinc finger BED domain-containing protein 5, hemoglobin subunit beta-2, integrator complex subunit 7, apolipoprotein A-I, and glutamyl aminopeptidase were differentially presented in the plasma of pregnant and non-pregnant female dogs. To the best of our knowledge, this is the first report on the plasma protein profile of pregnant and non-pregnant female dogs. In this study, we identified proteins that have not been previously identified in dogs. Our findings showed that numerous protein spots were differentially presented in the plasma of female dogs during normal pregnancy. Although we identified only a limited number of differentially presented proteins, our study demonstrated that the plasma protein profile changed during pregnancy in female dogs, which suggests its importance in maintaining pregnancy. Further studies are necessary to define complete plasma protein profile of pregnant female dogs and to identify all proteins that are differentially presented in the pregnant animals compared with the non-pregnant ones. In addition, studies are warranted to explain the role of those proteins in maintaining the pregnancy and their usefulness in detection of early pregnancy

Integrating sustainable hunting in biodiversity protection in Central Africa: hot spots, weak spots, and strong spots.

Directory of Open Access Journals (Sweden)

John E Fa

Full Text Available Wild animals are a primary source of protein (bushmeat for people living in or near tropical forests. Ideally, the effect of bushmeat harvests should be monitored closely by making regular estimates of offtake rate and size of stock available for exploitation. However, in practice, this is possible in very few situations because it requires both of these aspects to be readily measurable, and even in the best case, entails very considerable time and effort. As alternative, in this study, we use high-resolution, environmental favorability models for terrestrial mammals (N = 165 in Central Africa to map areas of high species richness (hot spots and hunting susceptibility. Favorability models distinguish localities with environmental conditions that favor the species' existence from those with detrimental characteristics for its presence. We develop an index for assessing Potential Hunting Sustainability (PHS of each species based on their ecological characteristics (population density, habitat breadth, rarity and vulnerability, weighted according to restrictive and permissive assumptions of how species' characteristics are combined. Species are classified into five main hunting sustainability classes using fuzzy logic. Using the accumulated favorability values of all species, and their PHS values, we finally identify weak spots, defined as high diversity regions of especial hunting vulnerability for wildlife, as well as strong spots, defined as high diversity areas of high hunting sustainability potential. Our study uses relatively simple models that employ easily obtainable data of a species' ecological characteristics to assess the impacts of hunting in tropical regions. It provides information for management by charting the geography of where species are more or less likely to be at risk of extinction from hunting.

DRAMATIC CHANGE IN JUPITER'S GREAT RED SPOT FROM SPACECRAFT OBSERVATIONS

International Nuclear Information System (INIS)

Simon, Amy A.; Wong, Michael H.; De Pater, Imke; Rogers, John H.; Orton, Glenn S.; Carlson, Robert W.; Asay-Davis, Xylar; Marcus, Philip S.

2014-01-01

Jupiter's Great Red Spot (GRS) is one of its most distinct and enduring features. Since the advent of modern telescopes, keen observers have noted its appearance and documented a change in shape from very oblong to oval, confirmed in measurements from spacecraft data. It currently spans the smallest latitude and longitude size ever recorded. Here we show that this change has been accompanied by an increase in cloud/haze reflectance as sensed in methane gas absorption bands, increased absorption at wavelengths shorter than 500 nm, and increased spectral slope between 500 and 630 nm. These changes occurred between 2012 and 2014, without a significant change in internal tangential wind speeds; the decreased size results in a 3.2 day horizontal cloud circulation period, shorter than previously observed. As the GRS has narrowed in latitude, it interacts less with the jets flanking its north and south edges, perhaps allowing for less cloud mixing and longer UV irradiation of cloud and aerosol particles. Given its long life and observational record, we expect that future modeling of the GRS's changes, in concert with laboratory flow experiments, will drive our understanding of vortex evolution and stability in a confined flow field crucial for comparison with other planetary atmospheres

DRAMATIC CHANGE IN JUPITER'S GREAT RED SPOT FROM SPACECRAFT OBSERVATIONS

Energy Technology Data Exchange (ETDEWEB)

Simon, Amy A. [NASA Goddard Space Flight Center, 8800 Greenbelt Road, Code 690, Greenbelt, MD 20771 (United States); Wong, Michael H.; De Pater, Imke [Astronomy Department, University of California Berkeley, Berkeley, CA 94720 (United States); Rogers, John H. [British Astronomical Association, Burlington House, Piccadilly, London W1J 0DU (United Kingdom); Orton, Glenn S.; Carlson, Robert W. [Jet Propulsion Laboratory, California Institute of Technology, 4800 Oak Grove Drive, Pasadena, CA 91109 (United States); Asay-Davis, Xylar [Potsdam Institute for Climate Impact Research, Telegraphenberg A 31, 14473 Potsdam (Germany); Marcus, Philip S. [Department of Mechanical Engineering, University of California Berkeley, 6121 Etcheverry Hall, Mailstop 1740, Berkeley, CA 94720 (United States)

2014-12-20

Jupiter's Great Red Spot (GRS) is one of its most distinct and enduring features. Since the advent of modern telescopes, keen observers have noted its appearance and documented a change in shape from very oblong to oval, confirmed in measurements from spacecraft data. It currently spans the smallest latitude and longitude size ever recorded. Here we show that this change has been accompanied by an increase in cloud/haze reflectance as sensed in methane gas absorption bands, increased absorption at wavelengths shorter than 500 nm, and increased spectral slope between 500 and 630 nm. These changes occurred between 2012 and 2014, without a significant change in internal tangential wind speeds; the decreased size results in a 3.2 day horizontal cloud circulation period, shorter than previously observed. As the GRS has narrowed in latitude, it interacts less with the jets flanking its north and south edges, perhaps allowing for less cloud mixing and longer UV irradiation of cloud and aerosol particles. Given its long life and observational record, we expect that future modeling of the GRS's changes, in concert with laboratory flow experiments, will drive our understanding of vortex evolution and stability in a confined flow field crucial for comparison with other planetary atmospheres.

Bier spots

Directory of Open Access Journals (Sweden)

Ahu Yorulmaz,

2015-10-01

Full Text Available Also called as physiologic anemic macules, Bier spots are small, hypopigmented irregularly shaped macules against a background of diffuse erythema, which creates an appearance of speckled vascular mottling of the skin. Bier spots most commonly appear on distal portions of the limbs though there are case reports describing diffuse involvement, which also affect trunk and mucous membranes of the patient. Although the exact pathophysiological mechanisms underlying Bier spots still need to be elucidated, Bier spots have been suggested to be a vascular anomaly caused by vasoconstriction of small vessels. In addition, several diseases have been proposed to be associated with Bier spots, including scleroderma renal crisis, cryoglobulinemia, Peutz-Jeghers syndrome, alopecia areata and hypoplasia of the aorta, although it has not been shown whether these associations are casual or coincidental. The clinical presentation of Bier spots is quite typical. These tiny whitish macules easily become prominent when the affected limb is placed in a dependent position and fade away when the limb is raised. Here we report a case of Bier spots in a 32-year-old male patient with characteristical clinical manifestations.

Humoral immune response of the small-spotted catshark, Scyliorhinus canicula.

Science.gov (United States)

Crouch, Kathryn; Smith, Lauren E; Williams, Rebecca; Cao, Wei; Lee, Mike; Jensen, Allan; Dooley, Helen

2013-05-01

Cartilaginous fishes are the oldest group in which an adaptive immune system based on immunoglobulin-superfamily members is found. This manuscript compares humoral immune function in small-spotted catshark (Scyliorhinus canicula) with that described for spiny dogfish (Squalus acanthias), another member of the Squalomorphi superorder, and nurse shark, the model for humoral immunity in elasmobranchs and a member of the Galeomorphi superorder. Although small-spotted catshark and nurse shark are separated by over 200 million years we found that immunoglobulin isoforms are well conserved between the two species. However, the plasma protein profile of small-spotted catshark was most similar to that of spiny dogfish, with low levels of pentameric IgM, and IgNAR present as a multimer in plasma rather than a monomer. We show that an antigen-specific monomeric IgM response, with a profile similar to that described previously for nurse sharks, can be raised in small-spotted catshark. Lacking polyclonal or monoclonal antibody reagents for detecting catshark IgNAR we investigated phage-display and recombinant Fc-fusion protein expression as alternative methods to look for an antigen-specific response for this isotype. However, we could find no evidence of an antigen-specific IgNAR in the animals tested using either of these techniques. Thus, unlike nurse sharks where antigen-specific monomeric IgM and IgNAR appear together, it seems there may be a temporal or complete 'uncoupling' of these isotypes during a humoral response in the small-spotted catshark. Copyright © 2013 Elsevier Ltd. All rights reserved.

Robust model-based analysis of single-particle tracking experiments with Spot-On

Science.gov (United States)

Grimm, Jonathan B; Lavis, Luke D

2018-01-01

Single-particle tracking (SPT) has become an important method to bridge biochemistry and cell biology since it allows direct observation of protein binding and diffusion dynamics in live cells. However, accurately inferring information from SPT studies is challenging due to biases in both data analysis and experimental design. To address analysis bias, we introduce ‘Spot-On’, an intuitive web-interface. Spot-On implements a kinetic modeling framework that accounts for known biases, including molecules moving out-of-focus, and robustly infers diffusion constants and subpopulations from pooled single-molecule trajectories. To minimize inherent experimental biases, we implement and validate stroboscopic photo-activation SPT (spaSPT), which minimizes motion-blur bias and tracking errors. We validate Spot-On using experimentally realistic simulations and show that Spot-On outperforms other methods. We then apply Spot-On to spaSPT data from live mammalian cells spanning a wide range of nuclear dynamics and demonstrate that Spot-On consistently and robustly infers subpopulation fractions and diffusion constants. PMID:29300163

Changing lanes in a simulator: effects of aging on the control of the vehicle and visual inspection of mirrors and blind spot.

Science.gov (United States)

Lavalliere, Martin; Laurendeau, Denis; Simoneau, Martin; Teasdale, Normand

2011-04-01

The aim of this study was to examine lane change strategies in active younger and older drivers. Visual inspection of mirrors and the blind spot and the control of the vehicle were documented in a simulator environment. Younger (n = 10, 21-31 years) and older (n = 11, 65-75 years) active drivers drove through a continuous simulated environment including urban and rural sections. The scenario included events where, to negotiate a secure lane change, the driver needed to look at 3 regions of interest (ROI): (1) the rearview mirror, (2) the left side mirror, and (3) the left blind spot. The lane change maneuvers were necessary to avoid a vehicle parked halfway in the rightmost lane that was partially or completely blocking the lane or for overtaking a slower moving vehicle. Compared with younger drivers, older drivers showed a reduced frequency of visual inspection toward the rearview mirror and the blind spot. Also, though the older drivers showed a constant frequency of visual inspection across the 2 types of driving maneuvers, the younger drivers increased their frequency of inspection when overtaking a slower vehicle. Control of the car was mostly similar for both groups. A better knowledge of the drivers' visual search strategies when changing lanes could help in identifying suboptimal strategies at-risk of causing crashes and also serves to develop retraining programs.

Demography of Northern Spotted Owls in southwestern Oregon

Science.gov (United States)

Zabel, Cynthia J.; Salmons, Susan E.; Forsman, Eric D.; DeStefano, Stephen; Raphael, Martin G.; Gutierrez, R.J.

1996-01-01

Northern Spotted Owls (Strix occidentalis caurina) are associated with lower elevation, commercially valuable, late-successional coniferous forests in the Pacific Northwest. Meta-analyses of demographic parameters indicate that Northern Spotted Owl populations are declining throughout their range (Anderson and Burnham 1992, Burnham et al. this volume). Recent research has attempted to determine whether management activities have affected the viability of Spotted Owl populations, and results have led to development of conservation plans for the species (Dawson et al. 1987, Thomas et al. 1990, Murphy and Noon 1992, USDI 1992, Thomas et al. 1993b).In the Recovery Plan for the Northern Spotted Owl (USDI 1992b) threats to the species were identified as small population sizes, declining populations, limited amounts of habitat, continued loss and fragmentation of habitat, geographically isolated populations, and predation and competition from other avian species. Weather and fire are natural processes that also may affect reproductive success of Spotted Owls. Weather may be a factor in the high annual variability in fecundity of Spotted Owls, as has been suggested for other predatory bird species (Newton, 1979, 1986). However, these factors have not been addressed in previous studies of Spotted Owls.Our objectives were to estimate survival, fecundity, and annual rates of population change (l) for resident, territorial female Spotted Owls at two study areas in the coastal mountains of southwestern Oregon. We tested if the amount of rainfall was correlated with reproduction of Spotted Owls. While surveying for Spotted Owls, we documented the increased presence of Barred Owls (Strix varia), a potential competitor of Spotted Owls.

Formation of truncated proteins and high-molecular-mass aggregates upon soft illumination of photosynthetic proteins

DEFF Research Database (Denmark)

Rinalducci, Sara; Campostrini, Natascia; Antonioli, Paolo

2005-01-01

Different spot profiles were observed in 2D gel electrophoresis of thylakoid membranes performed either under complete darkness or by leaving the sample for a short time to low visible light. In the latter case, a large number of new spots with lower molecular masses, ranging between 15,000 and 25......,000 Da, were observed, and high-molecular-mass aggregates, seen as a smearing in the upper part of the gel, appeared in the region around 250 kDa. Identification of protein(s) contained in these new spots by MS/MS revealed that most of them are simply truncated proteins deriving from native ones...

The rise and fall of a human recombination hot spot.

Science.gov (United States)

Jeffreys, Alec J; Neumann, Rita

2009-05-01

Human meiotic crossovers mainly cluster into narrow hot spots that profoundly influence patterns of haplotype diversity and that may also affect genome instability and sequence evolution. Hot spots also seem to be ephemeral, but processes of hot-spot activation and their subsequent evolutionary dynamics remain unknown. We now analyze the life cycle of a recombination hot spot. Sperm typing revealed a polymorphic hot spot that was activated in cis by a single base change, providing evidence for a primary sequence determinant necessary, though not sufficient, to activate recombination. This activating mutation occurred roughly 70,000 y ago and has persisted to the present, most likely fortuitously through genetic drift despite its systematic elimination by biased gene conversion. Nonetheless, this self-destructive conversion will eventually lead to hot-spot extinction. These findings define a subclass of highly transient hot spots and highlight the importance of understanding hot-spot turnover and how it influences haplotype diversity.

Proteomic analysis of protein phosphatase Z1 from Candida albicans.

Directory of Open Access Journals (Sweden)

Bernadett Márkus

Full Text Available Protein phosphatase Z is a "novel type" fungus specific serine/threonine protein phosphatase. Previously our research group identified the CaPPZ1 gene in the opportunistic pathogen Candida albicans and reported that the gene deletion had several important physiological consequences. In order to reveal the protein targets and the associated mechanisms behind the functions of the phosphatase a proteomic method was adopted for the comparison of the cappz1 deletion mutant and the genetically matching QMY23 control strain. Proteins extracted from the control and deletion mutant strains were separated by two-dimensional gel electrophoresis and the protein spots were stained with RuBPS and Pro-Q Diamond in order to visualize the total proteome and the phosphoproteome, respectively. The alterations in spot intensities were determined by densitometry and were analysed with the Delta2D (Decodon software. Spots showing significantly different intensities between the mutant and control strains were excised from the gels and were digested with trypsin. The resulting peptides were identified by LC-MS/MS mass spectrometry. As many as 15 protein spots were found that exhibited significant changes in their intensity upon the deletion of the phosphatase and 20 phosphoproteins were identified in which the level of phosphorylation was modified significantly in the mutant. In agreement with previous findings we found that the affected proteins function in protein synthesis, oxidative stress response, regulation of morphology and metabolism. Among these proteins we identified two potential CaPpz1 substrates (Eft2 and Rpp0 that may regulate the elongation step of translation. RT-qPCR experiments revealed that the expression of the genes coding for the affected proteins was not altered significantly. Thus, the absence of CaPpz1 exerted its effects via protein synthesis/degradation and phosphorylation/dephosphorylation. In addition, our proteomics data strongly

Proteomic analysis of protein phosphatase Z1 from Candida albicans

Science.gov (United States)

Pfliegler, Walter P.; Petrényi, Katalin; Boros, Enikő; Pócsi, István; Tőzsér, József; Dombrádi, Viktor

2017-01-01

Protein phosphatase Z is a “novel type” fungus specific serine/threonine protein phosphatase. Previously our research group identified the CaPPZ1 gene in the opportunistic pathogen Candida albicans and reported that the gene deletion had several important physiological consequences. In order to reveal the protein targets and the associated mechanisms behind the functions of the phosphatase a proteomic method was adopted for the comparison of the cappz1 deletion mutant and the genetically matching QMY23 control strain. Proteins extracted from the control and deletion mutant strains were separated by two-dimensional gel electrophoresis and the protein spots were stained with RuBPS and Pro-Q Diamond in order to visualize the total proteome and the phosphoproteome, respectively. The alterations in spot intensities were determined by densitometry and were analysed with the Delta2D (Decodon) software. Spots showing significantly different intensities between the mutant and control strains were excised from the gels and were digested with trypsin. The resulting peptides were identified by LC-MS/MS mass spectrometry. As many as 15 protein spots were found that exhibited significant changes in their intensity upon the deletion of the phosphatase and 20 phosphoproteins were identified in which the level of phosphorylation was modified significantly in the mutant. In agreement with previous findings we found that the affected proteins function in protein synthesis, oxidative stress response, regulation of morphology and metabolism. Among these proteins we identified two potential CaPpz1 substrates (Eft2 and Rpp0) that may regulate the elongation step of translation. RT-qPCR experiments revealed that the expression of the genes coding for the affected proteins was not altered significantly. Thus, the absence of CaPpz1 exerted its effects via protein synthesis/degradation and phosphorylation/dephosphorylation. In addition, our proteomics data strongly suggested a role for

Structure-dependent phytotoxicity of catechins and other flavonoids: flavonoid conversions by cell-free protein extracts of Centaurea maculosa (spotted knapweed) roots.

Science.gov (United States)

Bais, Harsh Pal; Walker, Travis S; Kennan, Alan J; Stermitz, Frank R; Vivanco, Jorge M

2003-02-12

Invasive plants are believed to succeed in part by secretion of allelochemicals, thus displacing competing plant species. Centaurea maculosa (spotted knapweed) provides a classic example of this process. We have previously reported that spotted knapweed roots secrete (+/-)-catechin and that (-)-catechin, but not (+)-catechin, is phytotoxic and hence may be a major contributor to C. maculosa's invasive behavior in the rhizosphere. In this communication, we explore both structure/activity relationships for flavonoid phytotoxicity and possible biosynthetic pathways for root production of (+/-)-catechin. Kaempferol and dihydroquercetin were shown to be phytotoxic, while quercetin was not. Kaempferol was converted to dihydroquercetin and (+/-)-catechin when treated with total root protein extracts from C. maculosa, but quercetin was not. This finding suggests an alteration in the standard flavonoid biosynthetic pathway in C. maculosa roots, whereby kaempferol is not a dead-end product but serves as a precursor to dihydroquercetin, which in turn leads to (+/-)-catechin production.

Proteome Changes in Maize Embryo (Zea mays L) Induced by Ion Beam Implantation Treatment

International Nuclear Information System (INIS)

Li Yongliang; Qin Guangyong; Huo Yuping; Tian Shuangqi; Tang Jihua

2009-01-01

Low energy ion beam implantation was applied to the maize (Zea mays L) embryo proteome using two-dimensional gel electrophoresis. Protein profile analysis detected more than 1100 protein spots, 72 of which were determined to be expressed differently in the treated and control (not exposed to ion beam implantation) embryos. Of the 72 protein spots, 53 were up-regulated in the control and 19 were more abundantly expressed in the ion beam-treated embryos. The spots of up- or down-regulated proteins were identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Among the identified proteins, 11 were up-regulated in the treated embryos. Four of these up-regulated proteins were antioxidant molecules, three were related to stress response, two to sugar metabolism and two were associated with heat shock response. Of the five proteins up-regulated in the control embryos, three were functionally related to carbohydrate metabolism; the functions of the remaining two proteins were unknown. The data collected during this study indicate that treatment of maize embryos with low energy ion beam implantation induces changes in stress tolerance enzymes/proteins, possibly as a result of alterations in metabolism. (ion beam bioengineering)

Proteome Changes in Maize Embryo (Zea mays L) Induced by Ion Beam Implantation Treatment

Science.gov (United States)

Li, Yongliang; Tang, Jihua; Qin, Guangyong; Huo, Yuping; Tian, Shuangqi

2009-08-01

Low energy ion beam implantation was applied to the maize (Zea mays L) embryo proteome using two-dimensional gel electrophoresis. Protein profile analysis detected more than 1100 protein spots, 72 of which were determined to be expressed differently in the treated and control (not exposed to ion beam implantation) embryos. Of the 72 protein spots, 53 were up-regulated in the control and 19 were more abundantly expressed in the ion beam-treated embryos. The spots of up- or down-regulated proteins were identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Among the identified proteins, 11 were up-regulated in the treated embryos. Four of these up-regulated proteins were antioxidant molecules, three were related to stress response, two to sugar metabolism and two were associated with heat shock response. Of the five proteins up-regulated in the control embryos, three were functionally related to carbohydrate metabolism; the functions of the remaining two proteins were unknown. The data collected during this study indicate that treatment of maize embryos with low energy ion beam implantation induces changes in stress tolerance enzymes/proteins, possibly as a result of alterations in metabolism.

Proteomic analysis of Herbaspirillum seropedicae reveals ammonium-induced AmtB-dependent membrane sequestration of PII proteins.

Science.gov (United States)

Huergo, Luciano F; Noindorf, Lilian; Gimenes, Camila; Lemgruber, Renato S P; Cordellini, Daniela F; Falarz, Lucas J; Cruz, Leonardo M; Monteiro, Rose A; Pedrosa, Fábio O; Chubatsu, Leda S; Souza, Emanuel M; Steffens, Maria B R

2010-07-01

This study was aimed at describing the spectrum and dynamics of proteins associated with the membrane in the nitrogen-fixing bacterium Herbaspirillum seropedicae according to the availability of fixed nitrogen. Using two-dimensional electrophoresis we identified 79 protein spots representing 45 different proteins in the membrane fraction of H. seropedicae. Quantitative analysis of gel images of membrane extracts indicated two spots with increased levels when cells were grown under nitrogen limitation in comparison with nitrogen sufficiency; these spots were identified as the GlnK protein and as a conserved noncytoplasmic protein of unknown function which was encoded in an operon together with GlnK and AmtB. Comparison of gel images of membrane extracts from cells grown under nitrogen limitation or under the same regime but collected after an ammonium shock revealed two proteins, GlnB and GlnK, with increased levels after the shock. The P(II) proteins were not present in the membrane fraction of an amtB mutant. The results reported here suggest that changes in the cellular localization of P(II) might play a role in the control of nitrogen metabolism in H. seropedicae.

Transitional–turbulent spots and turbulent–turbulent spots in boundary layers

Science.gov (United States)

Wu, Xiaohua; Moin, Parviz; Wallace, James M.; Skarda, Jinhie; Lozano-Durán, Adrián; Hickey, Jean-Pierre

2017-01-01

Two observations drawn from a thoroughly validated direct numerical simulation of the canonical spatially developing, zero-pressure gradient, smooth, flat-plate boundary layer are presented here. The first is that, for bypass transition in the narrow sense defined herein, we found that the transitional–turbulent spot inception mechanism is analogous to the secondary instability of boundary-layer natural transition, namely a spanwise vortex filament becomes a Λ vortex and then, a hairpin packet. Long streak meandering does occur but usually when a streak is infected by a nearby existing transitional–turbulent spot. Streak waviness and breakdown are, therefore, not the mechanisms for the inception of transitional–turbulent spots found here. Rather, they only facilitate the growth and spreading of existing transitional–turbulent spots. The second observation is the discovery, in the inner layer of the developed turbulent boundary layer, of what we call turbulent–turbulent spots. These turbulent–turbulent spots are dense concentrations of small-scale vortices with high swirling strength originating from hairpin packets. Although structurally quite similar to the transitional–turbulent spots, these turbulent–turbulent spots are generated locally in the fully turbulent environment, and they are persistent with a systematic variation of detection threshold level. They exert indentation, segmentation, and termination on the viscous sublayer streaks, and they coincide with local concentrations of high levels of Reynolds shear stress, enstrophy, and temperature fluctuations. The sublayer streaks seem to be passive and are often simply the rims of the indentation pockets arising from the turbulent–turbulent spots. PMID:28630304

Transitional-turbulent spots and turbulent-turbulent spots in boundary layers.

Science.gov (United States)

Wu, Xiaohua; Moin, Parviz; Wallace, James M; Skarda, Jinhie; Lozano-Durán, Adrián; Hickey, Jean-Pierre

2017-07-03

Two observations drawn from a thoroughly validated direct numerical simulation of the canonical spatially developing, zero-pressure gradient, smooth, flat-plate boundary layer are presented here. The first is that, for bypass transition in the narrow sense defined herein, we found that the transitional-turbulent spot inception mechanism is analogous to the secondary instability of boundary-layer natural transition, namely a spanwise vortex filament becomes a [Formula: see text] vortex and then, a hairpin packet. Long streak meandering does occur but usually when a streak is infected by a nearby existing transitional-turbulent spot. Streak waviness and breakdown are, therefore, not the mechanisms for the inception of transitional-turbulent spots found here. Rather, they only facilitate the growth and spreading of existing transitional-turbulent spots. The second observation is the discovery, in the inner layer of the developed turbulent boundary layer, of what we call turbulent-turbulent spots. These turbulent-turbulent spots are dense concentrations of small-scale vortices with high swirling strength originating from hairpin packets. Although structurally quite similar to the transitional-turbulent spots, these turbulent-turbulent spots are generated locally in the fully turbulent environment, and they are persistent with a systematic variation of detection threshold level. They exert indentation, segmentation, and termination on the viscous sublayer streaks, and they coincide with local concentrations of high levels of Reynolds shear stress, enstrophy, and temperature fluctuations. The sublayer streaks seem to be passive and are often simply the rims of the indentation pockets arising from the turbulent-turbulent spots.

Studies of the viral binding proteins of shrimp BP53, a receptor of white spot syndrome virus.

Science.gov (United States)

Li, Chen; Gao, Xiao-Xiao; Huang, Jie; Liang, Yan

2016-02-01

The specific binding between viral attachment proteins (VAPs) of a virus and its cellular receptors on host cells mediates virus entry into host cells, which triggers subsequent viral infections. Previous studies indicate that F1 ATP synthase β subunit (named BP53), is found on the surface of shrimp cells and involved in white spot syndrome virus (WSSV) infection by functioning as a potential viral receptor. Herein, in a far-western blotting assay, three WSSV proteins with molecular weights of 28 kDa, 37 kDa, and >50 kDa were found to interact with BP53. The 28 kDa and 37 kDa proteins were identified as the envelope protein VP28 and VP37 of WSSV respectively, which could be recognized by the polyclonal antibodies. Enzyme-linked immunosorbent binding assays revealed that VP37 contributed to almost 80% of the binding capability for BP53 compared with the same amount of total WSSV protein. The relationship between BP53 and its complementary interacting protein, VP37, was visualized using a co-localization assay. Bound VP37 on the cell surface co-localized with BP53 and shared a similar subcellular location on the outer surface of shrimp cells. Pearson's correlation coefficients reached to 0.67 ± 0.05 and the Mander's overlap coefficients reached 0.70 ± 0.05, which indicated a strong relationship between the localization of BP53 and bound rVP37. This provides evidence for an interaction between BP53 and VP37 obtained at the molecular and cellular levels, supporting the hypothesis that BP53 serves as a receptor for WSSV by binding to VP37. The identification of the viral binding proteins of shrimp BP53 is helpful for better understanding the pathogenic mechanisms of WSSV to infect shrimp at the cellular level. Copyright © 2016 Elsevier Inc. All rights reserved.

The evolutionary turnover of recombination hot spots contributes to speciation in mice.

Science.gov (United States)

Smagulova, Fatima; Brick, Kevin; Pu, Yongmei; Camerini-Otero, R Daniel; Petukhova, Galina V

2016-02-01

Meiotic recombination is required for the segregation of homologous chromosomes and is essential for fertility. In most mammals, the DNA double-strand breaks (DSBs) that initiate meiotic recombination are directed to a subset of genomic loci (hot spots) by sequence-specific binding of the PRDM9 protein. Rapid evolution of the DNA-binding specificity of PRDM9 and gradual erosion of PRDM9-binding sites by gene conversion will alter the recombination landscape over time. To better understand the evolutionary turnover of recombination hot spots and its consequences, we mapped DSB hot spots in four major subspecies of Mus musculus with different Prdm9 alleles and in their F1 hybrids. We found that hot spot erosion governs the preferential usage of some Prdm9 alleles over others in hybrid mice and increases sequence diversity specifically at hot spots that become active in the hybrids. As crossovers are disfavored at such hot spots, we propose that sequence divergence generated by hot spot turnover may create an impediment for recombination in hybrids, potentially leading to reduced fertility and, eventually, speciation. Published by Cold Spring Harbor Laboratory Press.

Protection of Penaeus monodon against white spot syndrome virus using a WSSV subunit vaccine

NARCIS (Netherlands)

Witteveldt, J.; Vlak, J.M.; Hulten, van M.C.W.

2004-01-01

Although invertebrates lack a true adaptive immune response, the potential to vaccinate Penaeus monodon shrimp against white spot syndrome virus (WSSV) using the WSSV envelope proteins VP19 and VP28 was evaluated. Both structural WSSV proteins were N-terminally fused to the maltose binding protein

Simplified quantification of urinary protein excretion in children with nephrotic syndrome

International Nuclear Information System (INIS)

Mustafa, G.; Khan, P.A.; Hussain, Z.; Iqbal, M.

2007-01-01

To assess the value of single voided random (spot) urinary protein to creatinine ratio in accurately predicting the 24-hour urinary protein excretion in Pakistani pediatric population with nephrotic syndrome. Fifty seven children between 1-18 years with nephrotic syndrome were included. Seventy pairs of spot urine (5 milliliter) and 24-hour urine were collected in different phases of their disease e.g. initial, induction and remission. The protein to creatinine ratio was determined in spot urine samples and total protein content in 24-hour urine samples. The correlation between the ratio and 24-hour urinary protein excreted was determined using Pearson's coefficient (r) linear regression analysis. The protein to creatinine ratio in a spot urine sample was significantly correlated with the 24-hour urinary protein. The correlation coefficient (least square method) was found to be significant (r=0.9444). A random (spot) urinary protein to creatinine ratio of greater than 2 correlated well with the massive proteinuria (i.e. nephrotic syndrome), between 2 to 0.2 indicated glomerulopathy while a ratio of less than 0.2 was suggestive of physiological values. The random spot urinary protein to creatinine ratio can reliably be used to assess the degree of proteinuria in children with nephrotic syndrome and can replace the 24-hour urinary protein excretion/collection. (author)

Spotted inflation

International Nuclear Information System (INIS)

Matsuda, Tomohiro

2010-01-01

We describe new scenarios for generating curvature perturbations when inflaton (curvaton) has significant interactions. We consider a ''spot'', which arises from interactions associated with an enhanced symmetric point (ESP) on the trajectory. Our first example uses the spot to induce a gap in the field equation. We observe that the gap in the field equation may cause generation of curvature perturbation if it does not appear simultaneous in space. The mechanism is similar to the scenario of inhomogeneous phase transition. Then we observe that the spot interactions may initiate warm inflation in the cold Universe. Creation of cosmological perturbation is discussed in relation to the inflaton dynamics and the modulation associated with the spot interactions

Spot Weight Adaptation for Moving Target in Spot Scanning Proton Therapy.

Science.gov (United States)

Morel, Paul; Wu, Xiaodong; Blin, Guillaume; Vialette, Stéphane; Flynn, Ryan; Hyer, Daniel; Wang, Dongxu

2015-01-01

This study describes a real-time spot weight adaptation method in spot-scanning proton therapy for moving target or moving patient, so that the resultant dose distribution closely matches the planned dose distribution. The method proposed in this study adapts the weight (MU) of the delivering pencil beam to that of the target spot; it will actually hit during patient/target motion. The target spot that a certain delivering pencil beam may hit relies on patient monitoring and/or motion modeling using four-dimensional (4D) CT. After the adapted delivery, the required total weight [Monitor Unit (MU)] for this target spot is then subtracted from the planned value. With continuous patient motion and continuous spot scanning, the planned doses to all target spots will eventually be all fulfilled. In a proof-of-principle test, a lung case was presented with realistic temporal and motion parameters; the resultant dose distribution using spot weight adaptation was compared to that without using this method. The impact of the real-time patient/target position tracking or prediction was also investigated. For moderate motion (i.e., mean amplitude 0.5 cm), D95% to the planning target volume (PTV) was only 81.5% of the prescription (RX) dose; with spot weight adaptation PTV D95% achieves 97.7% RX. For large motion amplitude (i.e., 1.5 cm), without spot weight adaptation PTV D95% is only 42.9% of RX; with spot weight adaptation, PTV D95% achieves 97.7% RX. Larger errors in patient/target position tracking or prediction led to worse final target coverage; an error of 3 mm or smaller in patient/target position tracking is preferred. The proposed spot weight adaptation method was able to deliver the planned dose distribution and maintain target coverage when patient motion was involved. The successful implementation of this method would rely on accurate monitoring or prediction of patient/target motion.

Spot Weight Adaptation for Moving Target in Spot Scanning Proton Therapy

Directory of Open Access Journals (Sweden)

Paul eMorel

2015-05-01

Full Text Available Purpose: This study describes a real-time spot weight adaptation method in spot-scanning proton therapy for moving target or moving patient, so that the resultant dose distribution closely matches the planned dose distribution. Materials and Methods: The method proposed in this study adapts the weight (MU of the delivering pencil beam to that of the target spot it will actually hit during patient/target motion. The target spot a certain delivering pencil beam may hit relies on patient monitoring and/or motion modeling using four-dimensional (4D CT. After the adapted delivery, the required total weight (MU for this target spot is then subtracted from the planned value. With continuous patient motion and continuous spot scanning, the planned doses to all target spots will eventually be all fulfilled. In a proof-of-principle test, a lung case was presented with realistic temporal and motion parameters; the resultant dose distribution using spot weight adaptation was compared to that without using this method. The impact of the real-time patient/target position tracking or prediction was also investigated.Results: For moderate motion (i.e., mean amplitude 0.5 cm, D95% to the planning target volume (PTV was only 81.5% of the prescription (RX dose; with spot weight adaptation PTV D95% achieves 97.7%RX. For large motion amplitude (i.e., 1.5 cm, without spot weight adaptation PTV D95% is only 42.9% of RX; with spot weight adaptation, PTV D95% achieves 97.7%RX. Larger errors in patient/target position tracking or prediction led to worse final target coverage; an error of 3mm or smaller in patient/target position tracking is preferred. Conclusion: The proposed spot weight adaptation method was able to deliver the planned dose distribution and maintain target coverage when patient motion was involved. The successful implementation of this method would rely on accurate monitoring or prediction of patient/target motion.

Nitrate as a probe of cytochrome c surface: crystallographic identification of crucial "hot spots" for protein-protein recognition.

Science.gov (United States)

De March, Matteo; Demitri, Nicola; De Zorzi, Rita; Casini, Angela; Gabbiani, Chiara; Guerri, Annalisa; Messori, Luigi; Geremia, Silvano

2014-06-01

The electrostatic surface of cytochrome c and its changes with the iron oxidation state are involved in the docking and undocking processes of this protein to its biological partners in the mitochondrial respiratory pathway. To investigate the subtle mechanisms of formation of productive macromolecular complexes and of their breakage following the electron transfer process, the X-ray structures of horse heart ferri-cytochrome c (trigonal form) and ferro-cytochrome c (monoclinic form) were obtained using nitrate ions both as a crystallizing agent and an anionic probe for mapping the electrostatic surface changes. Both crystal forms contain three protein molecules in the asymmetric unit. In addition, a total of 21.5 and 18 crystallographically independent nitrate ions were identified for the trigonal and monoclinic forms, respectively. By matching all the six crystallographically independent protein molecules, 26 different anion-protein interaction sites were identified on the surfaces of cytochrome c, 10 of which were found in both forms, 8 present only in the oxidized and 8 only in the reduced form. The structural analysis of the electron transfer complexes, based on this new information, suggests a specific exit strategy for cytochrome c after formation of productive protein-protein complexes: a directional sliding mechanism for the electron shuttle on the surface of the redox partner is proposed to take place after the electron transfer process has occurred. Copyright © 2014 Elsevier Inc. All rights reserved.

Bier spots

OpenAIRE

Ahu Yorulmaz,; Seray Kulcu Cakmak; Esra Ar?; Ferda Artuz

2015-01-01

Also called as physiologic anemic macules, Bier spots are small, hypopigmented irregularly shaped macules against a background of diffuse erythema, which creates an appearance of speckled vascular mottling of the skin. Bier spots most commonly appear on distal portions of the limbs though there are case reports describing diffuse involvement, which also affect trunk and mucous membranes of the patient. Although the exact pathophysiological mechanisms underlying Bier spots still need to be elu...

Judging a salmon by its spots: environmental variation is the primary determinant of spot patterns in Salmo salar.

Science.gov (United States)

Jørgensen, Katarina M; Solberg, Monica F; Besnier, Francois; Thorsen, Anders; Fjelldal, Per Gunnar; Skaala, Øystein; Malde, Ketil; Glover, Kevin A

2018-04-12

In fish, morphological colour changes occur from variations in pigment concentrations and in the morphology, density, and distribution of chromatophores in the skin. However, the underlying mechanisms remain unresolved in most species. Here, we describe the first investigation into the genetic and environmental basis of spot pattern development in one of the world's most studied fishes, the Atlantic salmon. We reared 920 salmon from 64 families of domesticated, F1-hybrid and wild origin in two contrasting environments (Hatchery; tanks for the freshwater stage and sea cages for the marine stage, and River; a natural river for the freshwater stage and tanks for the marine stage). Fish were measured, photographed and spot patterns evaluated. In the Hatchery experiment, significant but modest differences in spot density were observed among domesticated, F1-hybrid (1.4-fold spottier than domesticated) and wild salmon (1.7-fold spottier than domesticated). A heritability of 6% was calculated for spot density, and a significant QTL on linkage group SSA014 was detected. In the River experiment, significant but modest differences in spot density were also observed among domesticated, F1-hybrid (1.2-fold spottier than domesticated) and wild salmon (1.8-fold spottier than domesticated). Domesticated salmon were sevenfold spottier in the Hatchery vs. River experiment. While different wild populations were used for the two experiments, on average, these were 6.2-fold spottier in the Hatchery vs. River experiment. Fish in the Hatchery experiment displayed scattered to random spot patterns while fish in the River experiment displayed clustered spot patterns. These data demonstrate that while genetics plays an underlying role, environmental variation represents the primary determinant of spot pattern development in Atlantic salmon.

A spatial analysis of population dynamics and climate change in Africa: potential vulnerability hot spots emerge where precipitation declines and demographic pressures coincide

Science.gov (United States)

López-Carr, David; Pricope, Narcisa G.; Aukema, Juliann E.; Jankowska, Marta M.; Funk, Christopher C.; Husak, Gregory J.; Michaelsen, Joel C.

2014-01-01

We present an integrative measure of exposure and sensitivity components of vulnerability to climatic and demographic change for the African continent in order to identify “hot spots” of high potential population vulnerability. Getis-Ord Gi* spatial clustering analyses reveal statistically significant locations of spatio-temporal precipitation decline coinciding with high population density and increase. Statistically significant areas are evident, particularly across central, southern, and eastern Africa. The highly populated Lake Victoria basin emerges as a particularly salient hot spot. People located in the regions highlighted in this analysis suffer exceptionally high exposure to negative climate change impacts (as populations increase on lands with decreasing rainfall). Results may help inform further hot spot mapping and related research on demographic vulnerabilities to climate change. Results may also inform more suitable geographical targeting of policy interventions across the continent.

Spot weld arrangement effects on the fatigue behavior of multi-spot welded joints

International Nuclear Information System (INIS)

Hassanifard, Soran; Zehsaz, Mohammad; Esmaeili, Firooz

2011-01-01

In the present study, the effects of spot weld arrangements in multi-spot welded joints on the fatigue behavior of the joints are studied. Three different four-spot welded joints are considered: one-row four-spot parallel to the loading direction, one-row four-spot perpendicular to the loading direction and two-row four-spot weld specimens. The experimental fatigue test results reveal that the differences between the fatigue lives of three spot welded types in the low cycle regime are more considerable than those in the high cycle regime. However, all kinds of spot weld specimens have similar fatigue strength when approaching a million cycles. A non-linear finite element analysis is performed to obtain the relative stress gradients, effective distances and notch strength reduction factors based on the volumetric approach. The work here shows that the volumetric approach does a very good job in predicting the fatigue life of the multi-spot welded joints

Proteomic Comparison between Maturation Drying and Prematurely Imposed Drying of Zea mays Seeds Reveals a Potential Role of Maturation Drying in Preparing Proteins for Seed Germination, Seedling Vigor, and Pathogen Resistance

DEFF Research Database (Denmark)

Wang, Wei-Qing; Ye, Jian-Qing; Rogowska-Wrzesinska, Adelina

2014-01-01

We have studied the role(s) of maturation drying in the acquisition of germinability, seedling vigor and pathogen resistance by comparing the proteome changes in maize embryo and endosperm during mature and prematurely imposed drying. Prematurely imposed dried seeds at 40 days after pollination...... (DAP) germinated almost as well as mature seeds (at 65 DAP), but their seedling growth was slower and they were seriously infected by fungi. A total of 80 and 114 proteins were identified to change at least two-fold (p ... abundant in this group and may contribute to the acquisition of seed germinability. However, a relatively large number of proteins changed in the embryo (47 spots) and endosperm (76 spots) specifically during maturation drying. Among these proteins, storage proteins in the embryo and defense proteins...

Energy is not Coffee. An assessment of blind spots on energy spot-markets

International Nuclear Information System (INIS)

Jepma, C.J.; Spijker, E.; Van der Gaast, W.; De Jong, F.; Overmars, P.

2006-01-01

This study was to be the first in a series of studies on the title subject. It specifically focuses on the differences and similarities with a number of other spot-markets and aims to frame the energy spot markets and their potential development into a broader perspective. Main conclusion is that energy spot-markets differ from several other physical and non-physical spot-markets in many ways. This implies that 'perfect' energy spot-markets may inherently be (much) less perfect than other spot-markets that have approximated the stage of theoretical perfection

Hot Spots in a Network of Functional Sites

Science.gov (United States)

Ozbek, Pemra; Soner, Seren; Haliloglu, Turkan

2013-01-01

It is of significant interest to understand how proteins interact, which holds the key phenomenon in biological functions. Using dynamic fluctuations in high frequency modes, we show that the Gaussian Network Model (GNM) predicts hot spot residues with success rates ranging between S 8–58%, C 84–95%, P 5–19% and A 81–92% on unbound structures and S 8–51%, C 97–99%, P 14–50%, A 94–97% on complex structures for sensitivity, specificity, precision and accuracy, respectively. High specificity and accuracy rates with a single property on unbound protein structures suggest that hot spots are predefined in the dynamics of unbound structures and forming the binding core of interfaces, whereas the prediction of other functional residues with similar dynamic behavior explains the lower precision values. The latter is demonstrated with the case studies; ubiquitin, hen egg-white lysozyme and M2 proton channel. The dynamic fluctuations suggest a pseudo network of residues with high frequency fluctuations, which could be plausible for the mechanism of biological interactions and allosteric regulation. PMID:24023934

[Rocky Mountain spotted fever in Brazil].

Science.gov (United States)

del Sá DelFiol, Fernando; Junqueira, Fábio Miranda; da Rocha, Maria Carolina Pereira; de Toledo, Maria Inês; Filho, Silvio Barberato

2010-06-01

Although the number of confirmed cases of spotted fever has been declining in Brazil since 2005, the mortality rate (20% to 30%) is still high in comparison to other countries. This high mortality rate is closely related to the difficulty in making the diagnosis and starting the correct treatment. Only two groups of antibiotics have proven clinical effectiveness against spotted fever: chloramphenicol and tetracyclines. Until recently, the use of tetracyclines was restricted to adults because of the associated bone and tooth changes in children. Recently, however, the American Academy of Pediatrics and various researchers have recommended the use of doxycycline in children. In more severe cases, chloramphenicol injections are often preferred in Brazil because of the lack of experience with injectable tetracycline. Since early diagnosis and the adequate drug treatment are key to a good prognosis, health care professionals must be better prepared to recognize and treat spotted fever.

HotSpot Wizard 3.0: web server for automated design of mutations and smart libraries based on sequence input information.

Science.gov (United States)

Sumbalova, Lenka; Stourac, Jan; Martinek, Tomas; Bednar, David; Damborsky, Jiri

2018-05-23

HotSpot Wizard is a web server used for the automated identification of hotspots in semi-rational protein design to give improved protein stability, catalytic activity, substrate specificity and enantioselectivity. Since there are three orders of magnitude fewer protein structures than sequences in bioinformatic databases, the major limitation to the usability of previous versions was the requirement for the protein structure to be a compulsory input for the calculation. HotSpot Wizard 3.0 now accepts the protein sequence as input data. The protein structure for the query sequence is obtained either from eight repositories of homology models or is modeled using Modeller and I-Tasser. The quality of the models is then evaluated using three quality assessment tools-WHAT_CHECK, PROCHECK and MolProbity. During follow-up analyses, the system automatically warns the users whenever they attempt to redesign poorly predicted parts of their homology models. The second main limitation of HotSpot Wizard's predictions is that it identifies suitable positions for mutagenesis, but does not provide any reliable advice on particular substitutions. A new module for the estimation of thermodynamic stabilities using the Rosetta and FoldX suites has been introduced which prevents destabilizing mutations among pre-selected variants entering experimental testing. HotSpot Wizard is freely available at http://loschmidt.chemi.muni.cz/hotspotwizard.

Identification of proteins from cambium tissues of the chinese white poplar (populus tomentosa) sampled during the growing season

International Nuclear Information System (INIS)

Xie, J.; Liu, S.; Qi, Q.; Hou, Y.

2014-01-01

Various protein extraction methods have been used to investigate Chinese white poplar (Populus tomentosa) proteomics. However, extracting and characterizing proteins from woody plants remains a challenge. Two-dimensional gel electrophoresis is a powerful, widely used method for the analysis of complex protein mixtures extracted from biological samples. The technique separates mixtures of proteins along two dimensions, by isoelectric point and molecular weight, and can resolve thousands of different proteins. Here, we report a new application of two-dimensional gel electrophoresis to investigate the proteomics of P. tomentosa cambium tissues over the course of a growing season. Of three protein extraction methods that we compared (the Tris-phenol method, trichloroacetic acid-acetone method, and trichloroacetic acid-acetone-phenol method), trichloroacetic acid-acetone was the most efficient approach for protein extraction from cambium tissues of P. tomentosa. After extraction, the proteins were separated using two-dimensional gel electrophoresis. The protein quantities of six spots changed over the course of the growing season from February to July. Five spots were identified using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry, and the sixth spot was identified by liquid chromatography-mass spectrometry. The proteins included enolase, class Ia chitinase, and four unnamed proteins. Our results show the best approach to proteomics in P. tomentosa and reveal trends in protein activities during a growing season in this tree species. (author)

A novel white spot syndrome virus protein WSSV164 controls prophenoloxidases, PmproPOs in shrimp melanization cascade.

Science.gov (United States)

Sangsuriya, Pakkakul; Charoensapsri, Walaiporn; Sutthangkul, Jantiwan; Senapin, Saengchan; Hirono, Ikuo; Tassanakajon, Anchalee; Amparyup, Piti

2018-09-01

Melanization, mediated by the prophenoloxidase (proPO)-activating system, is an important innate immune response in invertebrates. The implication of the proPO system in antiviral response and the suppression of host proPO activation by the viral protein have previously been demonstrated in shrimp. However, the molecular mechanism of viral-host interactions in the proPO cascade remains largely unexplored. Here, we characterized the viral protein, namely, WSSV164, which was initially identified from the forward suppression subtractive hybridization (SSH) cDNA library of the PmproPO1/2 co-silenced black tiger shrimp Penaeus monodon that was challenged with white spot syndrome virus (WSSV). Using the yeast two-hybrid system, WSSV164 was found to interact with the PmproPO2 protein. The subsequent validation assay by co-immunoprecipitation revealed that WSSV164 directly bound to both PmproPO1 and PmproPO2. The gene silencing experiment was carried out to explore the role of WSSV164 in the control of the proPO pathway in shrimp, and the results showed that suppression of WSSV164 can restore PO activity in WSSV-infected shrimp hemolymph. The recombinant proteins of PmproPO1 and PmproPO2 were produced in Sf-9 cells and were shown to be successfully activated by exogenous trypsin and endogenous serine proteinases from shrimp hemocyte lysate supernatant (HLS), yielding PO activity in vitro. Moreover, the activated PO activity in shrimp HLS was dose-dependently reduced by the recombinant WSSV164 protein, suggesting that WSSV164 may interfere with the activation of the proPO system in shrimp. Taken together, these results suggest an alternative infection route of WSSV through the encoded viral protein WSSV164 that binds to the PmproPO1 and PmproPO2 proteins, interfering with the activation of the melanization cascade in shrimp. Copyright © 2018 Elsevier Ltd. All rights reserved.

The NSs protein of tomato spotted wilt virus is required for persistent infection and transmission by Frankliniella occidentalis.

Science.gov (United States)

Margaria, P; Bosco, L; Vallino, M; Ciuffo, M; Mautino, G C; Tavella, L; Turina, M

2014-05-01

Tomato spotted wilt virus (TSWV) is the type member of tospoviruses (genus Tospovirus), plant-infecting viruses that cause severe damage to ornamental and vegetable crops. Tospoviruses are transmitted by thrips in the circulative propagative mode. We generated a collection of NSs-defective TSWV isolates and showed that TSWV coding for truncated NSs protein could not be transmitted by Frankliniella occidentalis. Quantitative reverse transcription (RT)-PCR and immunostaining of individual insects detected the mutant virus in second-instar larvae and adult insects, demonstrating that insects could acquire and accumulate the NSs-defective virus. Nevertheless, adults carried a significantly lower viral load, resulting in the absence of transmission. Genome sequencing and analyses of reassortant isolates showed genetic evidence of the association between the loss of competence in transmission and the mutation in the NSs coding sequence. Our findings offer new insight into the TSWV-thrips interaction and Tospovirus pathogenesis and highlight, for the first time in the Bunyaviridae family, a major role for the S segment, and specifically for the NSs protein, in virulence and efficient infection in insect vector individuals. Our work is the first to show a role for the NSs protein in virus accumulation in the insect vector in the Bunyaviridae family: demonstration was obtained for the system TSWV-F. occidentalis, arguably one of the most damaging combination for vegetable crops. Genetic evidence of the involvement of the NSs protein in vector transmission was provided with multiple approaches.

Quantitative characterization of conformational-specific protein-DNA binding using a dual-spectral interferometric imaging biosensor

Science.gov (United States)

Zhang, Xirui; Daaboul, George G.; Spuhler, Philipp S.; Dröge, Peter; Ünlü, M. Selim

2016-03-01

DNA-binding proteins play crucial roles in the maintenance and functions of the genome and yet, their specific binding mechanisms are not fully understood. Recently, it was discovered that DNA-binding proteins recognize specific binding sites to carry out their functions through an indirect readout mechanism by recognizing and capturing DNA conformational flexibility and deformation. High-throughput DNA microarray-based methods that provide large-scale protein-DNA binding information have shown effective and comprehensive analysis of protein-DNA binding affinities, but do not provide information of DNA conformational changes in specific protein-DNA complexes. Building on the high-throughput capability of DNA microarrays, we demonstrate a quantitative approach that simultaneously measures the amount of protein binding to DNA and nanometer-scale DNA conformational change induced by protein binding in a microarray format. Both measurements rely on spectral interferometry on a layered substrate using a single optical instrument in two distinct modalities. In the first modality, we quantitate the amount of binding of protein to surface-immobilized DNA in each DNA spot using a label-free spectral reflectivity technique that accurately measures the surface densities of protein and DNA accumulated on the substrate. In the second modality, for each DNA spot, we simultaneously measure DNA conformational change using a fluorescence vertical sectioning technique that determines average axial height of fluorophores tagged to specific nucleotides of the surface-immobilized DNA. The approach presented in this paper, when combined with current high-throughput DNA microarray-based technologies, has the potential to serve as a rapid and simple method for quantitative and large-scale characterization of conformational specific protein-DNA interactions.DNA-binding proteins play crucial roles in the maintenance and functions of the genome and yet, their specific binding mechanisms are

Mass Spectrometry Method to Measure Membrane Proteins in Dried Blood Spots for the Detection of Blood Doping Practices in Sport.

Science.gov (United States)

Cox, Holly D; Eichner, Daniel

2017-09-19

The dried blood spot (DBS) matrix has significant utility for applications in the field where venous blood collection and timely shipment of labile blood samples is difficult. Unfortunately, protein measurement in DBS is hindered by high abundance proteins and matrix interference that increases with hematocrit. We developed a DBS method to enrich for membrane proteins and remove soluble proteins and matrix interference. Following a wash in a series of buffers, the membrane proteins are digested with trypsin and quantitated by parallel reaction monitoring mass spectrometry methods. The DBS method was applied to the quantification of four cell-specific cluster of differentiation (CD) proteins used to count cells by flow cytometry, band 3 (CD233), CD71, CD45, and CD41. We demonstrate that the DBS method counts low abundance cell types such as immature reticulocytes as well as high abundance cell types such as red blood cells, white blood cells, and platelets. When tested in 82 individuals, counts obtained by the DBS method demonstrated good agreement with flow cytometry and automated hematology analyzers. Importantly, the method allows longitudinal monitoring of CD protein concentration and calculation of interindividual variation which is difficult by other methods. Interindividual variation of band 3 and CD45 was low, 6 and 8%, respectively, while variation of CD41 and CD71 was higher, 18 and 78%, respectively. Longitudinal measurement of CD71 concentration in DBS over an 8-week period demonstrated intraindividual variation 17.1-38.7%. Thus, the method may allow stable longitudinal measurement of blood parameters currently monitored to detect blood doping practices.

Modeling Hot-Spot Contributions in Shocked High Explosives at the Mesoscale

Energy Technology Data Exchange (ETDEWEB)

Harrier, Danielle [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

2015-08-12

When looking at performance of high explosives, the defects within the explosive become very important. Plastic bonded explosives, or PBXs, contain voids of air and bonder between the particles of explosive material that aid in the ignition of the explosive. These voids collapse in high pressure shock conditions, which leads to the formation of hot spots. Hot spots are localized high temperature and high pressure regions that cause significant changes in the way the explosive material detonates. Previously hot spots have been overlooked with modeling, but now scientists are realizing their importance and new modeling systems that can accurately model hot spots are underway.

Changes in protein expression in the sheep abomasum following trickle infection with Teladorsagia circumcincta.

Science.gov (United States)

Pemberton, Alan D; Brown, Jeremy K; Craig, Nicky M; Pate, Judith; McLean, Kevin; Inglis, Neil F; Knox, David; Knight, Pamela A

2012-03-01

Continual low-level exposure of sheep to the helminth Teladorsagia circumcincta elicits a temporary protective immunity, where factors in the immune abomasal mucosa prevent penetration of infective larvae, but which is essentially lost within 6 weeks of cessation of parasite challenge. Here, a proteomic approach was used to identify proteins that are differentially regulated in immune compared to naïve sheep, as potential key mediators of immunity. Six naïve sheep and 12 sheep trickle-infected with T. circumcincta were treated with anthelmintic, and the naïve (control) and 6 immune sheep were killed 7 days later. The remaining 6 sheep (immune waning) were killed 42 days after anthelmintic treatment. Abomasal tissue samples were subjected to 2D-gel electrophoresis and densitometric analysis. Selected spots (n=73) were identified by peptide mass fingerprinting and confirmatory Western blotting was carried out for 10 proteins. Spots selectively up-regulated in immune versus control, but not immune waning versus control sheep, included galectin-15 and thioredoxin, which were confirmed by Western blotting. In immune sheep, serum albumin was significantly down-regulated and albumin proteolytic cleavage fragments were increased compared to controls. Unexpectedly, albumin mRNA was relatively highly expressed in control mucosa, down-regulated in immune, and was immunolocalized to mucus-producing epithelial cells. Thus we have identified differential expression of a number of proteins following T. circumcincta trickle infection that may play a role in host protection and inhibition of parasite establishment.

Radiation induced changes in proteome of mice jejunum: an in vivo 2DE study

International Nuclear Information System (INIS)

Bajaj, Sania; Dutta, Ajaswrata; Gupta, Manju L.

2014-01-01

Radiation exposure results in severe damage to biological system, by affecting cellular macromolecules of an individual. Damage to bio-molecules may lead to up/down-regulation of proteins, leading to dysfunction of organs. Gastrointestinal tract a key organ for digestion, absorption and barrier to the luminal bacteria and toxins, is one of the highly sensitive radiosensitive organ. Current study is focused on differential proteomic approach to understand the effect of radiation on intestinal (jejunum) proteins in a time dependent manner. Experiments were carried out initially to determine the appropriate conditions for separation of proteins in GI tissue of non irradiated control male C57BL6/J mice. 8-10 weeks old animals were exposed to 9 Gy (lethal) dose of gamma radiation. Differential expression of gastrointestinal tissue (jejunum) proteome was studied by 2DE at different time intervals. The intensity of protein spots of different treatment groups and control was measured by PD Quest software and the differential expression of respective proteins was calculated manually. Comparison of 2-DE gel images of irradiated jejunum tissue samples showed differential expression of various proteins when compared with untreated samples. A significant upregulation of total protein spots was observed within 1 hr group of 9 Gy radiation exposed sample and maximum down-regulation was evident at 72 hr. Out of 24 spots identified in the irradiated samples, 15 spots were down-regulated, and 3 spots were found missing in 72 hr group of irradiated samples respectively. Time dependent regulation of protein expression in irradiated jejunum was thus prominently evident. The data obtained from the present study has revealed differential radio sensitivity of some of the proteins which certainly have a definite role in inducing major cellular changes after radiation exposure. The finding also suggests that proteomic approach could be a potential tool to access the role of specific

Dietary protein to maximize resistance training: a review and examination of protein spread and change theories.

Science.gov (United States)

Bosse, John D; Dixon, Brian M

2012-09-08

An appreciable volume of human clinical data supports increased dietary protein for greater gains from resistance training, but not all findings are in agreement. We recently proposed "protein spread theory" and "protein change theory" in an effort to explain discrepancies in the response to increased dietary protein in weight management interventions. The present review aimed to extend "protein spread theory" and "protein change theory" to studies examining the effects of protein on resistance training induced muscle and strength gains. Protein spread theory proposed that there must have been a sufficient spread or % difference in g/kg/day protein intake between groups during a protein intervention to see muscle and strength differences. Protein change theory postulated that for the higher protein group, there must be a sufficient change from baseline g/kg/day protein intake to during study g/kg/day protein intake to see muscle and strength benefits. Seventeen studies met inclusion criteria. In studies where a higher protein intervention was deemed successful there was, on average, a 66.1% g/kg/day between group intake spread versus a 10.2% g/kg/day spread in studies where a higher protein diet was no more effective than control. The average change in habitual protein intake in studies showing higher protein to be more effective than control was +59.5% compared to +6.5% when additional protein was no more effective than control. The magnitudes of difference between the mean spreads and changes of the present review are similar to our previous review on these theories in a weight management context. Providing sufficient deviation from habitual intake appears to be an important factor in determining the success of additional protein in enhancing muscle and strength gains from resistance training. An increase in dietary protein favorably effects muscle and strength during resistance training.

Redox proteomics changes in the fungal pathogen Trichosporon asahii on arsenic exposure: identification of protein responses to metal-induced oxidative stress in an environmentally-sampled isolate.

Directory of Open Access Journals (Sweden)

Sidra Ilyas

Full Text Available Trichosporon asahii is a yeast pathogen implicated in opportunistic infections. Cultures of an isolate collected from industrial wastewater were exposed for 2 days to 100 mg/L sodium arsenite (NaAsO2 and cadmium (CdCl2. Both metals reduced glutathione transferase (GST activity but had no effect on superoxide dismutase or catalase. NaAsO2 exposure increased glutathione reductase activity while CdCl2 had no effect. Protein thiols were labeled with 5-iodoacetamido fluorescein followed by one dimensional electrophoresis which revealed extensive protein thiol oxidation in response to CdCl2 treatment but thiol reduction in response to NaAsO2. Two dimensional electrophoresis analyses showed that the intensity of some protein spots was enhanced on treatment as judged by SameSpots image analysis software. In addition, some spots showed decreased IAF fluorescence suggesting thiol oxidation. Selected spots were excised and tryptic digested for identification by MALDI-TOF/TOF MS. Twenty unique T. asahii proteins were identified of which the following proteins were up-regulated in response to NaAsO2: 3-isopropylmalate dehydrogenase, phospholipase B, alanine-glyoxylate aminotransferase, ATP synthase alpha chain, 20S proteasome beta-type subunit Pre3p and the hypothetical proteins A1Q1_08001, A1Q2_03020, A1Q1_06950, A1Q1_06913. In addition, the following showed decreased thiol-associated fluorescence consistent with thiol oxidation; aconitase; aldehyde reductase I; phosphoglycerate kinase; translation elongation factor 2; heat shock protein 70 and hypothetical protein A1Q2_04745. Some proteins showed both increase in abundance coupled with decrease in IAF fluorescence; 3-hydroxyisobutyryl-CoA hydrolase; homoserine dehydrogenase Hom6 and hypothetical proteins A1Q2_03020 and A1Q1_00754. Targets implicated in redox response included 10 unique metabolic enzymes, heat shock proteins, a component of the 20S proteasome and translation elongation factor 2. These data

Oil prices, speculation, and fundamentals. Interpreting causal relations among spot and futures prices

International Nuclear Information System (INIS)

Kaufmann, Robert K.; Ullman, Ben

2009-01-01

A consensus that the world oil market is unified begs the question, where do innovations in oil prices enter the market? Here we investigate where changes in the price of crude oil originate and how they spread by examining causal relationships among prices for crude oils from North America, Europe, Africa, and the Middle East on both spot and futures markets. Results indicate that innovations first appear in spot prices for Dubai-Fateh and spread to other spot and futures prices while other innovations first appear in the far month contract for West Texas Intermediate and spread to other exchanges and contracts. Links between spot and futures markets are relatively weak and this may have allowed the long-run relationship between spot and future prices to change after September 2004. Together, these results suggest that market fundamentals initiated a long-term increase in oil prices that was exacerbated by speculators, who recognized an increase in the probability that oil prices would rise over time. (author)

Serum proteomic changes after randomized prolonged erythropoietin treatment and/or endurance training: detection of novel biomarkers.

Directory of Open Access Journals (Sweden)

Britt Christensen

Full Text Available Despite implementation of the biological passport to detect erythropoietin abuse, a need for additional biomarkers remains. We used a proteomic approach to identify novel serum biomarkers of prolonged erythropoiesis-stimulating agent (ESA exposure (Darbepoietin-α and/or aerobic training.Thirty-six healthy young males were randomly assigned to the following groups: Sedentary-placebo (n = 9, Sedentary-ESA (n = 9, Training-placebo (n = 10, or Training-ESA (n = 8. They were treated with placebo/Darbepoietin-α subcutaneously once/week for 10 weeks followed by a 3-week washout period. Training consisted of supervised biking 3/week for 13 weeks at the highest possible intensity. Serum was collected at baseline, week 3 (high dose Darbepoietin-α, week 10 (reduced dose Darbepoietin-α, and after a 3-week washout period.Serum proteins were separated according to charge and molecular mass (2D-gel electrophoresis. The identity of proteins from spots exhibiting altered intensity was determined by mass spectrometry.Six protein spots changed in response to Darbepoietin-α treatment. Comparing all 4 experimental groups, two protein spots (serotransferrin and haptoglobin/haptoglobin related protein showed a significant response to Darbepoietin-α treatment. The haptoglobin/haptoglobin related protein spot showed a significantly lower intensity in all subjects in the training-ESA group during the treatment period and increased during the washout period.An isoform of haptoglobin/haptoglobin related protein could be a new anti-doping marker and merits further research.ClinicalTrials.gov NCT01320449.

Rocky Mountain spotted fever

Science.gov (United States)

... spotted fever on the foot Rocky Mountain spotted fever, petechial rash Antibodies Deer and dog tick References McElligott SC, Kihiczak GG, Schwartz RA. Rocky Mountain spotted fever and other rickettsial infections. In: Lebwohl MG, Heymann ...

Changes in protein expression in testes of L2 strain Taiwan country chickens in response to acute heat stress.

Science.gov (United States)

Wang, Shih-Han; Cheng, Chuen-Yu; Chen, Chao-Jung; Chen, Hsin-Hsin; Tang, Pin-Chi; Chen, Chih-Feng; Lee, Yen-Pai; Huang, San-Yuan

2014-07-01

Heat stress causes a decrease of fertility in roosters. Yet, the way acute heat stress affects protein expression remains poorly understood. This study investigated differential protein expression in testes of the L2 strain of Taiwan country chickens following acute heat stress. Twelve 45-week-old roosters were allocated into four groups, including control roosters kept at 25 °C, roosters subjected to 38 °C acute heat stress for 4 hours without recovery, with 2 hours of recovery, and with 6 hours of recovery. Testis samples were collected for morphologic assay and protein analysis. Some of the differentially expressed proteins were validated by Western blot and immunohistochemistry. Abnormal and apoptotic spermatogenic cells were observed at 2 hours of recovery after acute heat stress, especially among the spermatocytes. Two-dimensional difference gel electrophoresis revealed that 119 protein spots were differentially expressed in chicken testes following heat stress, and peptide mass fingerprinting revealed that these spots contained 92 distinct proteins. In the heat-stressed samples, the heat shock proteins, chaperonin containing t-complex, and proteasome subunits were downregulated, and glutathione S-transferase, transgelin, and DJ-1 were upregulated. Our results demonstrate that acute heat stress impairs the processes of translation, protein folding, and protein degradation, and thus results in apoptosis and interferes with spermatogenesis. On the other hand, the increased expression of antioxidant enzymes, including glutathione S-transferase and DJ-1, may attenuate heat-induced damage. These findings may have implications for breeding chickens that can tolerate more extreme conditions. Copyright © 2014 Elsevier Inc. All rights reserved.

Protein-Protein Docking in Drug Design and Discovery.

Science.gov (United States)

Kaczor, Agnieszka A; Bartuzi, Damian; Stępniewski, Tomasz Maciej; Matosiuk, Dariusz; Selent, Jana

2018-01-01

Protein-protein interactions (PPIs) are responsible for a number of key physiological processes in the living cells and underlie the pathomechanism of many diseases. Nowadays, along with the concept of so-called "hot spots" in protein-protein interactions, which are well-defined interface regions responsible for most of the binding energy, these interfaces can be targeted with modulators. In order to apply structure-based design techniques to design PPIs modulators, a three-dimensional structure of protein complex has to be available. In this context in silico approaches, in particular protein-protein docking, are a valuable complement to experimental methods for elucidating 3D structure of protein complexes. Protein-protein docking is easy to use and does not require significant computer resources and time (in contrast to molecular dynamics) and it results in 3D structure of a protein complex (in contrast to sequence-based methods of predicting binding interfaces). However, protein-protein docking cannot address all the aspects of protein dynamics, in particular the global conformational changes during protein complex formation. In spite of this fact, protein-protein docking is widely used to model complexes of water-soluble proteins and less commonly to predict structures of transmembrane protein assemblies, including dimers and oligomers of G protein-coupled receptors (GPCRs). In this chapter we review the principles of protein-protein docking, available algorithms and software and discuss the recent examples, benefits, and drawbacks of protein-protein docking application to water-soluble proteins, membrane anchoring and transmembrane proteins, including GPCRs.

The Spotting Distribution of Wildfires

Directory of Open Access Journals (Sweden)

Jonathan Martin

2016-06-01

Full Text Available In wildfire science, spotting refers to non-local creation of new fires, due to downwind ignition of brands launched from a primary fire. Spotting is often mentioned as being one of the most difficult problems for wildfire management, because of its unpredictable nature. Since spotting is a stochastic process, it makes sense to talk about a probability distribution for spotting, which we call the spotting distribution. Given a location ahead of the fire front, we would like to know how likely is it to observe a spot fire at that location in the next few minutes. The aim of this paper is to introduce a detailed procedure to find the spotting distribution. Most prior modelling has focused on the maximum spotting distance, or on physical subprocesses. We will use mathematical modelling, which is based on detailed physical processes, to derive a spotting distribution. We discuss the use and measurement of this spotting distribution in fire spread, fire management and fire breaching. The appendix of this paper contains a comprehensive review of the relevant underlying physical sub-processes of fire plumes, launching fire brands, wind transport, falling and terminal velocity, combustion during transport, and ignition upon landing.

Evaluation of different protein extraction methods for banana (Musa spp.) root proteome analysis by two-dimensional electrophoresis.

Science.gov (United States)

Vaganan, M Mayil; Sarumathi, S; Nandakumar, A; Ravi, I; Mustaffa, M M

2015-02-01

Four protocols viz., the trichloroacetic acid-acetone (TCA), phenol-ammonium acetate (PAA), phenol/SDS-ammonium acetate (PSA) and trisbase-acetone (TBA) were evaluated with modifications for protein extraction from banana (Grand Naine) roots, considered as recalcitrant tissues for proteomic analysis. The two-dimensional electrophoresis (2-DE) separated proteins were compared based on protein yield, number of resolved proteins, sum of spot quantity, average spot intensity and proteins resolved in 4-7 pI range. The PAA protocol yielded more proteins (0.89 mg/g of tissues) and protein spots (584) in 2-DE gel than TCA and other protocols. Also, the PAA protocol was superior in terms of sum of total spot quantity and average spot intensity than TCA and other protocols, suggesting phenol as extractant and ammonium acetate as precipitant of proteins were the most suitable for banana rooteomics analysis by 2-DE. In addition, 1:3 ratios of root tissue to extraction buffer and overnight protein precipitation were most efficient to obtain maximum protein yield.

Motion Interplay as a Function of Patient Parameters and Spot Size in Spot Scanning Proton Therapy for Lung Cancer

Science.gov (United States)

Grassberger, Clemens; Dowdell, Stephen; Lomax, Antony; Sharp, Greg; Shackleford, James; Choi, Noah; Willers, Henning; Paganetti, Harald

2013-01-01

Purpose Quantify the impact of respiratory motion on the treatment of lung tumors with spot scanning proton therapy. Methods and Materials 4D Monte Carlo simulations were used to assess the interplay effect, which results from relative motion of the tumor and the proton beam, on the dose distribution in the patient. Ten patients with varying tumor sizes (2.6-82.3cc) and motion amplitudes (3-30mm) were included in the study. We investigated the impact of the spot size, which varies between proton facilities, and studied single fractions and conventionally fractionated treatments. The following metrics were used in the analysis: minimum/maximum/mean dose, target dose homogeneity and 2-year local control rate (2y-LC). Results Respiratory motion reduces the target dose homogeneity, with the largest effects observed for the highest motion amplitudes. Smaller spot sizes (σ≈3mm) are inherently more sensitive to motion, decreasing target dose homogeneity on average by a factor ~2.8 compared to a larger spot size (σ≈13mm). Using a smaller spot size to treat a tumor with 30mm motion amplitude reduces the minimum dose to 44.7% of the prescribed dose, decreasing modeled 2y-LC from 87.0% to 2.7%, assuming a single fraction. Conventional fractionation partly mitigates this reduction, yielding a 2y-LC of 71.6%. For the large spot size, conventional fractionation increases target dose homogeneity and prevents a deterioration of 2y-LC for all patients. No correlation with tumor volume is observed. The effect on the normal lung dose distribution is minimal: observed changes in mean lung dose and lung V20 are interplay using a large spot size and conventional fractionation. For treatments employing smaller spot sizes and/or in the delivery of single fractions, interplay effects can lead to significant deterioration of the dose distribution and lower 2y-LC. PMID:23462423

Motion Interplay as a Function of Patient Parameters and Spot Size in Spot Scanning Proton Therapy for Lung Cancer

International Nuclear Information System (INIS)

Grassberger, Clemens; Dowdell, Stephen; Lomax, Antony; Sharp, Greg; Shackleford, James; Choi, Noah; Willers, Henning; Paganetti, Harald

2013-01-01

Purpose: To quantify the impact of respiratory motion on the treatment of lung tumors with spot scanning proton therapy. Methods and Materials: Four-dimensional Monte Carlo simulations were used to assess the interplay effect, which results from relative motion of the tumor and the proton beam, on the dose distribution in the patient. Ten patients with varying tumor sizes (2.6-82.3 cc) and motion amplitudes (3-30 mm) were included in the study. We investigated the impact of the spot size, which varies between proton facilities, and studied single fractions and conventionally fractionated treatments. The following metrics were used in the analysis: minimum/maximum/mean dose, target dose homogeneity, and 2-year local control rate (2y-LC). Results: Respiratory motion reduces the target dose homogeneity, with the largest effects observed for the highest motion amplitudes. Smaller spot sizes (σ ≈ 3 mm) are inherently more sensitive to motion, decreasing target dose homogeneity on average by a factor 2.8 compared with a larger spot size (σ ≈ 13 mm). Using a smaller spot size to treat a tumor with 30-mm motion amplitude reduces the minimum dose to 44.7% of the prescribed dose, decreasing modeled 2y-LC from 87.0% to 2.7%, assuming a single fraction. Conventional fractionation partly mitigates this reduction, yielding a 2y-LC of 71.6%. For the large spot size, conventional fractionation increases target dose homogeneity and prevents a deterioration of 2y-LC for all patients. No correlation with tumor volume is observed. The effect on the normal lung dose distribution is minimal: observed changes in mean lung dose and lung V 20 are <0.6 Gy(RBE) and <1.7%, respectively. Conclusions: For the patients in this study, 2y-LC could be preserved in the presence of interplay using a large spot size and conventional fractionation. For treatments using smaller spot sizes and/or in the delivery of single fractions, interplay effects can lead to significant deterioration of the

Coral bleaching and ocean ''hot spots''

Energy Technology Data Exchange (ETDEWEB)

Goreau, T.J. (Global Coral Reef Alliance, Chappaqua, NY (United States)); Hayes, R.L. (Howard Univ., Washington, DC (United States). College of Medicine)

1994-05-01

Global sea-surface temperature maps show that mass coral-reef bleaching episodes between 1983 and 1991 followed positive anomalies more than 1 deg C above long-term monthly averages (''hot spots'') during the preceding warm season. Irregular formation, movement, and disappearance of hot spots make their detailed long-term prediction impossible, but they can be tracked in real time from satellite data. Monitoring of ocean hot spots and of coral bleaching is needed if the Framework Convention of Climate Change is to meet its goal of protecting the most temperature sensitive ecosystems. 47 refs, 3 figs

The critical spot eraser—a method to interactively control the correction of local hot and cold spots in IMRT planning

International Nuclear Information System (INIS)

Süss, Philipp; Bortz, Michael; Küfer, Karl-Heinz; Thieke, Christian

2013-01-01

Common problems in inverse radiotherapy planning are localized dose insufficiencies like hot spots in organs at risk or cold spots inside targets. These are hard to correct since the optimization is based on global evaluations like maximum/minimum doses, equivalent uniform doses or dose–volume constraints for whole structures. In this work, we present a new approach to locally correct the dose of any given treatment plan. Once a treatment plan has been found that is acceptable in general but requires local corrections, these areas are marked by the planner. Then the system generates new plans that fulfil the local dose goals. Consequently, it is possible to interactively explore all plans between the locally corrected plans and the original treatment plan, allowing one to exactly adjust the degree of local correction and how the plan changes overall. Both the amount (in Gy) and the size of the local dose change can be navigated. The method is introduced formally as a new mathematical optimization setting, and is evaluated using a clinical example of a meningioma at the base of the skull. It was possible to eliminate a hot spot outside the target volume while controlling the dose changes to all other parts of the treatment plan. The proposed method has the potential to become the final standard step of inverse treatment planning. For more information on this article, see medicalphysicsweb.org (paper)

Hematological changes in white spot syndrome virus-infected shrimp, Fenneropenaeus chinensis (Osbeck)

Science.gov (United States)

Feng, Shouming; Zhan, Wenbin; Xing, Jing; Li, Jun; Yang, Kai; Wang, Jing

2008-08-01

The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis.

SPOT Program

Science.gov (United States)

Smith, Jason T.; Welsh, Sam J.; Farinetti, Antonio L.; Wegner, Tim; Blakeslee, James; Deboeck, Toni F.; Dyer, Daniel; Corley, Bryan M.; Ollivierre, Jarmaine; Kramer, Leonard;

Monoclonal protein reference change value as determined by gel-based serum protein electrophoresis.

Science.gov (United States)

Salamatmanesh, Mina; McCudden, Christopher R; McCurdy, Arleigh; Booth, Ronald A

2018-01-01

The International Myeloma Working Group recommendations for monitoring disease progression or response include quantitation of the involved monoclonal immunoglobulin. They have defined the minimum change criteria of ≧25% with an absolute change of no gel-based serum protein electrophoresis. Sixteen clinically stable MGUS patients were identified from our clinical hematology database. Individual biological variability (CVi) was determined and used to calculate a monoclonal protein reference change value (RCV). Analytical variability of the normal protein fractions (albumin, alpha-1, alpha-2, beta, total gamma) ranged from 1.3% for albumin to 5.8% for the alpha-1 globulins. CVa of low (5.6g/L) and high (32.2g/L) concentration monoclonal proteins were 3.1% and 22.2%, respectively. Individual CVi of stable patients ranged from 3.5% to 24.5% with a CVi of 12.9%. The reference change value (RCV) at a 95% probability was determined to be 36.7% (low) 39.6% (high) using our CVa and CVi. Serial monitoring of monoclonal protein concentration is important for MGUS and multiple myeloma patients. Accurate criteria for interpreting a change in monoclonal protein concentration are required for appropriate decision making. We used QC results and real-world conditions to assess imprecision of serum protein fractions including low and high monoclonal protein fractions and clinically stable MGUS patients to determine CVi and RCV. The calculated RCVs of 36.7% (low) and 39.6% (high) in this study were greater that reported previously and greater than the established criteria for relapse. Response criteria may be reassessed to increase sensitivity and specificity for detection of response. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

[Effect of resin infiltration treatment on the colour of white spot lesions].

Science.gov (United States)

Zhao, Xiaoyi; Gao, Xuejun

2014-06-01

This study aimed to evaluate the effect of resin infiltration and fluoride solution on masking different demineralized white spot lesions by assessing color change. Artificial white spot lesions were produced on 60 human molars. Each sample had at least two enamel surfaces opened (named A and B). The samples were randomly divided into groups 1, 2, and 3 according to their time of demineralization (24, 48, and 72 h). After demineralization, the A spot of each sample was treated by resin infiltration. The B spot was treated with 0.1% fluoride solution daily for 30 days. After the remineralization of the B spot, resin infiltration was used again on the B spot of each sample. Color assessment was performed by a spectrophotometer in five distinct stages: baseline, after the production of artificial caries, after resin infiltration of A spots, after 30 days of fluoride solution treatment of B spots, and after resin infiltration of remineralized B spots. Before demineralization, the L* values of spots A and B in all groups were not significantly different (P > 0.05), whereas the L* values of spots A and B were significantly increased after demineralization. The L* values of A spots recovered significantly after treatment by resin infiltration (P 0.05) after fluoride treatment compared with that after demineralization. After resin infiltration on B spots, the L* values recovered but could not reach the baseline nor the level of A spots treated by resin infiltration only. Resin infiltration is a more effective treatment for masking white spot lesions than traditional fluoride treatment. The effect of masking white spot lesions has certain relationships with the degree of demineralization and activity of the lesion.

Spatio-temporal changes in germination and radical elongation of barley seeds tracked by proteome analysis of dissected embryo, aleurone layer and endosperm tissues

DEFF Research Database (Denmark)

Bønsager, Birgit Christine; Finnie, Christine; Roepstorff, P.

2007-01-01

proteins to appear (at 4 h PI). Other early changes were observed that affected spots containing desiccation stress-associated late embryogenesis abundant and abscisic acid (ABA)-induced proteins. From 12 h PI proteins characteristic for desiccation stress disappeared rapidly, as did a putative embryonic...

Effects of macromolecular crowding on protein conformational changes.

Directory of Open Access Journals (Sweden)

Hao Dong

2010-07-01

Full Text Available Many protein functions can be directly linked to conformational changes. Inside cells, the equilibria and transition rates between different conformations may be affected by macromolecular crowding. We have recently developed a new approach for modeling crowding effects, which enables an atomistic representation of "test" proteins. Here this approach is applied to study how crowding affects the equilibria and transition rates between open and closed conformations of seven proteins: yeast protein disulfide isomerase (yPDI, adenylate kinase (AdK, orotidine phosphate decarboxylase (ODCase, Trp repressor (TrpR, hemoglobin, DNA beta-glucosyltransferase, and Ap(4A hydrolase. For each protein, molecular dynamics simulations of the open and closed states are separately run. Representative open and closed conformations are then used to calculate the crowding-induced changes in chemical potential for the two states. The difference in chemical-potential change between the two states finally predicts the effects of crowding on the population ratio of the two states. Crowding is found to reduce the open population to various extents. In the presence of crowders with a 15 A radius and occupying 35% of volume, the open-to-closed population ratios of yPDI, AdK, ODCase and TrpR are reduced by 79%, 78%, 62% and 55%, respectively. The reductions for the remaining three proteins are 20-44%. As expected, the four proteins experiencing the stronger crowding effects are those with larger conformational changes between open and closed states (e.g., as measured by the change in radius of gyration. Larger proteins also tend to experience stronger crowding effects than smaller ones [e.g., comparing yPDI (480 residues and TrpR (98 residues]. The potentials of mean force along the open-closed reaction coordinate of apo and ligand-bound ODCase are altered by crowding, suggesting that transition rates are also affected. These quantitative results and qualitative trends will

Expression of Leaf Proteins in Two Cultivars of Bread Wheat under Cadmium and Mercury Stress Using Two-Dimensional Gel Electrophoresis

Directory of Open Access Journals (Sweden)

S. Y. Raeesi Sadati

2016-02-01

Full Text Available Wheat is an important source of human food. Cadmium and mercury bind to sulfhydryl groups of structural proteins and enzymes and cause inhibition in activity and decrease in protein production or interfere with the regulation of the enzymes. To study the effect of protein expression under different levels of cadmium and mercury, the experiment was conducted in a completely randomized design with three replications in Mohaghegh Ardabili University, Ardabil, Iran. Experimental factors consisted of two Gonbad and Tajan bread what cultivars, heavy metals in seven levels (four concentrations of mercuric chloride in 5, 10, 15 and 20 µM and cadmium chloride at two concentrations of 0.25 and 0.5 mM and sampling time after 8 and 16 hours of treatment. The Bradford method was used for quantitative analysis of proteins and 12% SDS-PAGE and two dimensional electrophorese techniques were hired for analysis of their expression. The results showed that under cadmium and mercury stresses, the total protein content increased compared to the control. Two-dimensional electrophoresis of proteins under cadmium stress showed differential expression of the protein spots on the plant leaves, than the control. In general, changes in the expression of proteins under the effect of cadmium stress were divided into two main categories: Spots 9, 10, 13, 14 and 16 belonged to proteins with reduced expression and the spots 1, 2, 8, 19 and 20 belonged to proteins with increased expression, in comparison to non-stressed control. These spots of up regulated proteins were directly related to the defense system against the heavy metal stress.

Experimental Study of Ignition by Hot Spot in Internal Combustion Engines

Science.gov (United States)

Serruys, Max

1938-01-01

In order to carry out the contemplated study, it was first necessary to provide hot spots in the combustion chamber, which could be measured and whose temperature could be changed. It seemed difficult to realize both conditions working solely on the temperature of the cooling water in a way so as to produce hot spots on the cylinder wall capable of provoking autoignition. Moreover, in the majority of practical cases, autoignition is produced by the spark plug, one of the least cooled parts in the engine. The first procedure therefore did not resemble that which most generally occurs in actual engine operation. All of these considerations caused us to reproduce similar hot spots at the spark plugs. The hot spots produced were of two kinds and designated with the name of thermo-electric spark plug and of metallic hot spot.

Progressive outer retinal necrosis presenting as cherry red spot.

Science.gov (United States)

Yiu, Glenn; Young, Lucy H

2012-10-01

To report a case of progressive outer retinal necrosis (PORN) presenting as a cherry red spot. Case report. A 53-year-old woman with recently diagnosed HIV and varicella-zoster virus (VZV) aseptic meningitis developed rapid sequential vision loss in both eyes over 2 months. Her exam showed a "cherry red spot" in both maculae with peripheral atrophy and pigmentary changes, consistent with PORN. Due to her late presentation and the rapid progression of her condition, she quickly developed end-stage vision loss in both eyes. PORN should be considered within the differential diagnosis of a "cherry red spot." Immune-deficient patients with a history of herpetic infection who present with visual loss warrant prompt ophthalmological evaluation.

Models of spots and flares

International Nuclear Information System (INIS)

Mullan, D.J.

1983-01-01

Laboratory experiments in recent years have shown that there are many more ways to drive a plasma out of equilibrium than to preserve equilibrium. In that sense, it is perhaps easier to understand why flares should occur in a stellar atmosphere than why a long-lived feature such as a dark spot should persist. The author summarizes work on the equilibrium structure of cool spots in the sun and stars. Since spots involve complex interactions between convective flows and magnetic fields, he needs to refer to observations for help in identifying the dominant processes which should enter into the modelling. His summary therefore begins by discussing certain relevant properties of spots in the solar atmosphere. The next sections deal with the magnetic fields in spots, the stability of spots, spot cooling and missing flux. The author concludes that spots should be viewed not simply as cool areas, but rather as engines which do the work of converting the energy of convective flows into flare-compatible form. (Auth.)

Nitrate as a probe of cytochrome c surface : crystallographic identification of crucial "hot spots" for protein-protein recognition

NARCIS (Netherlands)

De March, Matteo; Demitri, Nicola; De Zorzi, Rita; Casini, Angela; Gabbiani, Chiara; Guerri, Annalisa; Messori, Luigi; Geremia, Silvano

The electrostatic surface of cytochrome c and its changes with the iron oxidation state are involved in the docking and undocking processes of this protein to its biological partners in the mitochondrial respiratory pathway. To investigate the subtle mechanisms of formation of productive

Tibial dyschondroplasia-associated proteomic changes in chicken growth plate cartilage.

Science.gov (United States)

Rasaputra, K S; Liyanage, A B R; Lay, J O; McCarthy, F M; Rath, N C

2010-12-01

Tibial dyschondroplasia (TD) is a poultry leg problem that affects the proximal growth plate of the tibia, preventing its transition to bone. To understand the disease-induced proteomic changes, we compared the protein extracts of cartilage from normal and TD-affected growth plates. TD was induced by feeding thiram to chickens 2 wk before tissue harvest. Proteins were extracted from whole tissues and from conditioned media (CM) prepared by incubating appropriate growth plate tissues in serum-free culture medium for 48 hr. The extracts were prefractionated to contain proteins ranging between 10 and 100 kD. Equal amounts of proteins were subjected to 2D gel electrophoresis with three individual samples per group. The gels were silver stained, and digital images were compared and analyzed with Melanie software to determine differentially expressed protein spots. On comparison of two sets of gels, 47 matching spots were detected in tissue extracts and 27 in CM extracts. Among the matching spots, 12 were determined to be down-regulated in tissue extracts (P type II collagen precursor, and the expressed sequence tag pgm2n.pk014.f20, a protein with homology to human reticulocalbin-3 (RCN3). Most of the downregulated proteins are associated with signal transduction, energy metabolism, and secretory functions that are integral to cell viability. Consistent with our earlier findings that the TD chondrocytes are nonviable, the current results suggest that thiram very likely interferes with basic metabolic functions of chondrocytes, leading to their death and, consequently, to the pathogenesis of TD.

Utilizing Estimated Creatinine Excretion to Improve the Performance of Spot Urine Samples for the Determination of Proteinuria in Kidney Transplant Recipients.

Directory of Open Access Journals (Sweden)

Michael Ke Wang

Full Text Available Agreement between spot and 24-hour urine protein measurements is poor in kidney transplant recipients. We investigated whether using formulae to estimate creatinine excretion rate (eCER, rather than assuming a standard creatinine excretion rate, would improve the estimation of proteinuria from spot urine samples in kidney transplant recipients.We measured 24 hour urine protein and albumin and spot albumin:creatinine (ACR and spot protein:creatinine (PCR in 181 Kidney transplant recipients." We utilized 6 different published formulae (Fotheringham, CKD-EPI, Cockcroft-Gault, Walser, Goldwasser and Rule to estimate eCER and from it calculated estimated albumin and protein excretion rate (eAER and ePER. Bias, precision and accuracy (within 15%, 30% and 50% of ACR, PCR, eAER, ePER were compared to 24-hour urine protein and albumin.ACR and PCR significantly underestimated 24-hour albumin and protein excretion (ACR Bias (IQR, -5.9 mg/day; p< 0.01; PCR Bias, (IQR, -35.2 mg/day; p<0.01. None of the formulae used to calculate eAER or ePER had a bias that was significantly different from the 24-hour collection (eAER and ePER bias: Fotheringham -0.3 and 7.2, CKD-EPI 0.3 and 13.5, Cockcroft-Gault -3.2 and -13.9, Walser -1.7 and 3.1, Goldwasser -1.3 and -0.5, Rule -0.6 and 4.2 mg/day respectively. The accuracy for ACR and PCR were lower (within 30% being 38% and 43% respectively than the corresponding values estimated by utilizing eCER (for eAER 46% to 49% and ePER 46-54%.Utilizing estimated creatinine excretion to calculate eAER and ePER improves the estimation of 24-hour albuminuria/proteinuria with spot urine samples in kidney transplant recipients.

A PQL (protein quantity loci) analysis of mature pea seed proteins identifies loci determining seed protein composition.

Science.gov (United States)

Bourgeois, Michael; Jacquin, Françoise; Cassecuelle, Florence; Savois, Vincent; Belghazi, Maya; Aubert, Grégoire; Quillien, Laurence; Huart, Myriam; Marget, Pascal; Burstin, Judith

2011-05-01

Legume seeds are a major source of dietary proteins for humans and animals. Deciphering the genetic control of their accumulation is thus of primary significance towards their improvement. At first, we analysed the genetic variability of the pea seed proteome of three genotypes over 3 years of cultivation. This revealed that seed protein composition variability was under predominant genetic control, with as much as 60% of the spots varying quantitatively among the three genotypes. Then, by combining proteomic and quantitative trait loci (QTL) mapping approaches, we uncovered the genetic architecture of seed proteome variability. Protein quantity loci (PQL) were searched for 525 spots detected on 2-D gels obtained for 157 recombinant inbred lines. Most protein quantity loci mapped in clusters, suggesting that the accumulation of the major storage protein families was under the control of a limited number of loci. While convicilin accumulation was mainly under the control of cis-regulatory regions, vicilins and legumins were controlled by both cis- and trans-regulatory regions. Some loci controlled both seed protein composition and protein content and a locus on LGIIa appears to be a major regulator of protein composition and of protein in vitro digestibility. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hot spots of crop production changes at 1.5°C and 2°C

Science.gov (United States)

Schleussner, C. F.; Deryng, D.; Mueller, C.; Elliott, J. W.; Saeed, F.; Folberth, C.; Liu, W.; Wang, X.; Pugh, T.

2017-12-01

Studying changes in global and regional crop production is central for assessing the benefits of limiting global average temperature below 1.5ºC versus 2ºC. Projections of future climatic impacts on crop production are commonly focussed on focussing on mean changes. However, substantial risks are posed by extreme weather events such as heat waves and droughts that are of great relevance for imminent policy relevant questions such as price shocks or food security. Preliminary research on the benefits of keeping global average temperature increase below 1.5ºC versus 2ºC above pre-industrial levels has indicated that changes in extreme weather event occurrences will be more pronounced than changes in the mean climate. Here we will present results of crop yield projections for a set of global gridded crop models (GGCMs) for four major staple crops at 1.5°C and 2°C warming above pre-industrial levels using climate forcing data from the Half a degree Additional warming, Prognosis and Projected Impacts (HAPPI) project. We will assess changes in crop production on the global and regional level, and identify hot spots of change. The unique multi-ensemble setup allows to identify changes in extreme yield losses with multi-year to multi-decadal return periods, and thus elucidate the consequences for global and regional food security.

Effects of spot size and spot spacing on lateral penumbra reduction when using a dynamic collimation system for spot scanning proton therapy

International Nuclear Information System (INIS)

Hyer, Daniel E; Hill, Patrick M; Wang, Dongxu; Smith, Blake R; Flynn, Ryan T

2014-01-01

The purpose of this work was to investigate the reduction in lateral dose penumbra that can be achieved when using a dynamic collimation system (DCS) for spot scanning proton therapy as a function of two beam parameters: spot size and spot spacing. This is an important investigation as both values impact the achievable dose distribution and a wide range of values currently exist depending on delivery hardware. Treatment plans were created both with and without the DCS for in-air spot sizes (σ air ) of 3, 5, 7, and 9 mm as well as spot spacing intervals of 2, 4, 6 and 8 mm. Compared to un-collimated treatment plans, the plans created with the DCS yielded a reduction in the mean dose to normal tissue surrounding the target of 26.2–40.6% for spot sizes of 3–9 mm, respectively. Increasing the spot spacing resulted in a decrease in the time penalty associated with using the DCS that was approximately proportional to the reduction in the number of rows in the raster delivery pattern. We conclude that dose distributions achievable when using the DCS are comparable to those only attainable with much smaller initial spot sizes, suggesting that the goal of improving high dose conformity may be achieved by either utilizing a DCS or by improving beam line optics. (note)

On the origin of delta spots

International Nuclear Information System (INIS)

Tang, F.

1983-01-01

Mount Wilson sunspot drawings from 1966 through 1980 were used in conjunction with Hα filtergrams from Big Bear Solar Observatory to examine the origin of delta spots, spots with bipolar umbrae within one penumbra. Of the six cases we studied, five were formed by the union of non-paired spots. They are either shoved into one another by two neighboring growing bipoles or by a new spot born piggy-back style on an existing spot of opposite polarity. Proper motions of the growing spots take on curvilinear paths around one another to avoid a collision. This is the shear motion observed in delta spots (Tanaka, 1979). In the remaining case, the delta spot was formed by spots that emerged as a pair. Our findings indicate no intrinsic differences in the formation or the behavior between delta spots of normal magnetic configuration. (orig.)

Demographic response of northern spotted owls to barred owl removal

Science.gov (United States)

Diller, V. Lowell; Hamm, Keith A; Early, Desiree A; Lamphear, David W; Dugger, Katie M.; Yackulic, Charles B.; Schwarz, Carl J.; Carlson, Peter C.; McDonald, Trent L.

2016-01-01

the treated areas and 0.822 for the untreated areas. Derived estimates of population change on both study areas showed the same general decline before removal with an estimated slope of –0.0036 per year. Following removal, the rate of population change on the treated areas increased to an average of 1.029 but decreased to an average of 0.870 on the untreated areas. The results from this first experiment demonstrated that lethal removal of barred owls allowed the recovery of northern spotted owl populations in the treated portions of our study area. If additional federally funded barred owl removal experiments provide similar results, this could be the foundation for development of a long-term conservation strategy for northern spotted owls.

Protein expression in the nucleus accumbens of rats exposed to developmental vitamin D deficiency.

Directory of Open Access Journals (Sweden)

John McGrath

Full Text Available INTRODUCTION: Developmental vitamin D (DVD deficiency is a candidate risk factor for schizophrenia. Animal models have confirmed that DVD deficiency is associated with a range of altered genomic, proteomic, structural and behavioural outcomes in the rat. Because the nucleus accumbens has been implicated in neuropsychiatric disorders, in the current study we examined protein expression in this region in adult rats exposed to DVD deficiency METHODS: Female Sprague Dawley rats were maintained on a vitamin D deficient diet for 6 weeks, mated and allowed to give birth, after which a diet containing vitamin D was reintroduced. Male adult offspring (n = 8 were compared to control male (n = 8. 2-D gel electrophoresis-based proteomics and mass spectroscopy were used to investigate differential protein expression. RESULTS: There were 35 spots, mapped to 33 unique proteins, which were significantly different between the two groups. Of these, 22 were down-regulated and 13 up-regulated. The fold changes were uniformly small, with the largest FC being -1.67. Within the significantly different spots, three calcium binding proteins (calbindin1, calbindin2 and hippocalcin were altered. Other proteins associated with DVD deficiency related to mitochondrial function, and the dynamin-like proteins. CONCLUSIONS: Developmental vitamin D deficiency was associated with subtle changes in protein expression in the nucleus accumbens. Disruptions in pathways related to calcium-binding proteins and mitochondrial function may underlie some of the behavioural features associated with animal models of developmental vitamin D deficiency.

Proteomic and transcriptomic analysis of Arabidopsis seeds: molecular evidence for successive processing of seed proteins and its implication in the stress response to sulfur nutrition.

Science.gov (United States)

Higashi, Yasuhiro; Hirai, Masami Yokota; Fujiwara, Toru; Naito, Satoshi; Noji, Masaaki; Saito, Kazuki

2006-11-01

Seed storage proteins are synthesized as sources of carbon, nitrogen and sulfur for the next generation of plants. Their composition changes according to nutritional conditions. Here, we report the precise molecular identification of seed proteins by proteomic analysis of wild-type Arabidopsis thaliana and methionine-over-accumulating mutant mto1-1 plants. The identities of 50 protein spots were determined in the protein extract of mature Arabidopsis seeds by two-dimensional (2D) gel electrophoresis and subsequent mass spectrometric analysis. Of these protein spots, 42 were identified as derived from 12S globulins or 2S albumins. These results indicate that approximately 84% of protein species in Arabidopsis seeds are derived from a few genes coding for 12S globulins and 2S albumins. Extensive mass spectrometric analysis of the 42 spots revealed that successive C-terminal degradation occurred on the 12S globulins. The feasibility of this C-terminal processing was rationalized by molecular modeling of the three-dimensional structure of 12S globulins. The C-terminal degradation at glutamic acid residues of the 12S globulin subunits was repressed under sulfur-deficient conditions. Transcriptome analysis was combined with proteomic analysis to elucidate the mechanism of changes in seed protein composition in response to sulfur deficiency. The results suggest that seed storage proteins in Arabidopsis undergo multi-layer regulation, with emphasis on post-translational modifications that enable the plant to respond to sulfur deficiency.

Acute Phase Proteins and Variables of Protein Metabolism in Dairy Cows during the Pre- and Postpartal Period

Directory of Open Access Journals (Sweden)

Cs. Tóthová

2008-01-01

Full Text Available The objective of the present study was to compare the concentrations of acute phase proteins and selected variables of protein metabolism in dairy cows of the Slovak Spotted breed from 4 weeks before parturition to 10 weeks after parturition. Acute phase proteins - haptoglobin (Hp and serum amyloid A (SAA - and variables of protein metabolism - total proteins, albumin, urea, creatinine, total immunoglobulins - were evaluated in blood serum. Significant differences were found in average values of the Hp and SAA concentrations in several groups during the monitored period (P P P P P P P P P < 0.001. The above mentioned results indicate that in the time around parturition there are significant changes in concentrations of acute phase proteins, as well as in the whole protein metabolism of dairy cows. These facts suggest that the postparturient period is a critical biological phase, throughout which there is the highest incidence of metabolic disorders.

Rapid experimental SAD phasing and hot-spot identification with halogenated fragments

Energy Technology Data Exchange (ETDEWEB)

Bauman, Joseph D.; Harrison, Jerry Joe E. K.; Arnold, Eddy

2016-01-01

Through X-ray crystallographic fragment screening, 4-bromopyrazole was discovered to be a `magic bullet' that is capable of binding at many of the ligand `hot spots' found in HIV-1 reverse transcriptase (RT). The binding locations can be in pockets that are `hidden' in the unliganded crystal form, allowing rapid identification of these sites forin silicoscreening. In addition to hot-spot identification, this ubiquitous yet specific binding provides an avenue for X-ray crystallographic phase determination, which can be a significant bottleneck in the determination of the structures of novel proteins. The anomalous signal from 4-bromopyrazole or 4-iodopyrazole was sufficient to determine the structures of three proteins (HIV-1 RT, influenza A endonuclease and proteinase K) by single-wavelength anomalous dispersion (SAD) from single crystals. Both compounds are inexpensive, readily available, safe and very soluble in DMSO or water, allowing efficient soaking into crystals.

Rapid experimental SAD phasing and hot-spot identification with halogenated fragments

Directory of Open Access Journals (Sweden)

Joseph D. Bauman

2016-01-01

Full Text Available Through X-ray crystallographic fragment screening, 4-bromopyrazole was discovered to be a `magic bullet' that is capable of binding at many of the ligand `hot spots' found in HIV-1 reverse transcriptase (RT. The binding locations can be in pockets that are `hidden' in the unliganded crystal form, allowing rapid identification of these sites for in silico screening. In addition to hot-spot identification, this ubiquitous yet specific binding provides an avenue for X-ray crystallographic phase determination, which can be a significant bottleneck in the determination of the structures of novel proteins. The anomalous signal from 4-bromopyrazole or 4-iodopyrazole was sufficient to determine the structures of three proteins (HIV-1 RT, influenza A endonuclease and proteinase K by single-wavelength anomalous dispersion (SAD from single crystals. Both compounds are inexpensive, readily available, safe and very soluble in DMSO or water, allowing efficient soaking into crystals.

Spot market for uranium

International Nuclear Information System (INIS)

Colhoun, C.

1982-01-01

The spot market is always quoted for the price of uranium because little information is available about long-term contracts. A review of the development of spot market prices shows the same price curve swings that occur with all raw materials. Future long-term contracts will probably be lower to reflect spot market prices, which are currently in the real-value range of $30-$35. An upswing in the price of uranium could come in the next few months as utilities begin making purchases and trading from stockpiles. The US, unlike Europe and Japan, has already reached a supply and demand point where the spot market share is increasing. Forecasters cannot project the market price, they can only predict the presence of an oscillating spot or a secondary market. 5 figures

Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp

International Nuclear Information System (INIS)

Wang Yuzhen; Zhang Xiaohua; Yuan Li; Xu Tao; Rao Yu; Li Jia; Dai Heping

2008-01-01

White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP28 is one of the most important envelope proteins of WSSV. In this study, a recombinant antibody library, as single-chain fragment variable (scFv) format, displayed on phage was constructed using mRNA from spleen cells of mice immunized with full-length VP28 expressed in Escherichia coli. After several rounds of panning, six scFv antibodies specifically binding to the epitopes in the N-terminal, middle, and C-terminal regions of VP28, respectively, were isolated from the library. Using these scFv antibodies as tools, the epitopes in VP28 were located on the envelope of the virion by immuno-electron microscopy. Neutralization assay with these antibodies in vitro suggested that these epitopes may not be the attachment site of WSSV to host cell receptor. This study provides a new way to investigate the structure and function of the envelope proteins of WSSV

Immunoproteomic Analysis of the Excretory-Secretory Proteins from Spirometra Mansoni Sparganum

Directory of Open Access Journals (Sweden)

Zhong Quan Wang

2013-09-01

Full Text Available Background: Sparganosis is caused by the invasion of Spirometra sparganum into various tissues/organs. Subcutaneous sparganosis can be diagnosed by biopsy, while visceral/cerebral sparganosis is not easy to be diagnosed. The diagnosis de­pends largely on the detection of specific anti-sparganum antibodies. The specific­ity of the ELISA could be increased by using S. mansoni sparganum excretory–secre­tory (ES antigens, but it also had the cross-reactions with sera of patients with cysticercosis or paragonimiasis. The aim of this study was to identify early specific diagnostic antigens in S. mansoni sparganum ES proteins.Methods: The sparganum ES proteins were analyzed by two-dimensional electrophore­sis (2-DE and Western blot probed with early sera from infected mice at 14 days post-infection. The immunoreactive protein spots were characterized by MALDI-TOF/ TOF-MS.Results: A total of approximately 149 proteins spots were detected with isoelectric point (pI varying from 3 to 7.5 and molecular weight from 20 to 115 kDa and seven protein spots with molecular weight of 23-31 kDa were recognized by the infection sera. Three of seven spots were successfully identified and characterized as the same S. mansoni protein (cysteine protease, and the proteins of other 4 spots were not included in the databases.Conclusion: The cysteine protease from S. mansoni ES proteins recognized by early infection sera might be the early diagnostic antigens for sparganosis.

Proteomic analysis of halotolerant proteins under high and low salt stress in Dunaliella salina using two-dimensional differential in-gel electrophoresis

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Yan-Long Jia

2016-01-01

Full Text Available Abstract Dunaliella salina, a single-celled marine alga with extreme salt tolerance, is an important model organism for studying fundamental extremophile survival mechanisms and their potential practical applications. In this study, two-dimensional differential in-gel electrophoresis (2D-DIGE was used to investigate the expression of halotolerant proteins under high (3 M NaCl and low (0.75 M NaCl salt concentrations. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/TOF MS and bioinformatics were used to identify and characterize the differences among proteins. 2D-DIGE analysis revealed 141 protein spots that were significantly differentially expressed between the two salinities. Twenty-four differentially expressed protein spots were successfully identified by MALDI-TOF/TOF MS, including proteins in the following important categories: molecular chaperones, proteins involved in photosynthesis, proteins involved in respiration and proteins involved in amino acid synthesis. Expression levels of these proteins changed in response to the stress conditions, which suggests that they may be involved in the maintenance of intracellular osmotic pressure, cellular stress responses, physiological changes in metabolism, continuation of photosynthetic activity and other aspects of salt stress. The findings of this study enhance our understanding of the function and mechanisms of various proteins in salt stress.

A method for short term electricity spot price forecasting

International Nuclear Information System (INIS)

Koreneff, G.; Seppaelae, A.; Lehtonen, M.; Kekkonen, V.; Laitinen, E.; Haekli, J.; Antila, E.

1998-01-01

In Finland, the electricity market was de-regulated in November 1995. For the electricity purchase of power companies this has caused big changes, since the old tariff based contracts of bulk power supply have been replaced by negotiated bilateral short term contracts and by power purchase from the spot market. In the spot market, in turn, there are at the present two strong actors: The electricity exchange of Finland and the Nordic power pool which is run by the Swedish and Norwegian companies. Today, the power companies in Finland have short term trade with both of the electricity exchanges. The aim of this chapter is to present methods for spot price forecasting in the electricity exchange. The main focus is given to the Finnish circumstances. In the beginning of the presentation, the practices of the electricity exchange of Finland are described, and a brief presentation is given on the different contracts, or electricity products, available in the spot market. For comparison, the practices of the Nordic electricity exchange are also outlined. A time series technique for spot price forecasting is presented. The structure of the model is presented, and its validity is tested using real case data obtained from the Finnish power market. The spot price forecasting model is a part of a computer system for distribution energy management (DEM) in a de-regulated power market

A method for short term electricity spot price forecasting

Energy Technology Data Exchange (ETDEWEB)

Koreneff, G; Seppaelae, A; Lehtonen, M; Kekkonen, V [VTT Energy, Espoo (Finland); Laitinen, E; Haekli, J [Vaasa Univ. (Finland); Antila, E [ABB Transmit Oy (Finland)

1998-08-01

In Finland, the electricity market was de-regulated in November 1995. For the electricity purchase of power companies this has caused big changes, since the old tariff based contracts of bulk power supply have been replaced by negotiated bilateral short term contracts and by power purchase from the spot market. In the spot market, in turn, there are at the present two strong actors: The electricity exchange of Finland and the Nordic power pool which is run by the Swedish and Norwegian companies. Today, the power companies in Finland have short term trade with both of the electricity exchanges. The aim of this chapter is to present methods for spot price forecasting in the electricity exchange. The main focus is given to the Finnish circumstances. In the beginning of the presentation, the practices of the electricity exchange of Finland are described, and a brief presentation is given on the different contracts, or electricity products, available in the spot market. For comparison, the practices of the Nordic electricity exchange are also outlined. A time series technique for spot price forecasting is presented. The structure of the model is presented, and its validity is tested using real case data obtained from the Finnish power market. The spot price forecasting model is a part of a computer system for distribution energy management (DEM) in a de-regulated power market

Landuse change identification using SPOT 6 for food security analysis of Denpasar City, Bali Province, Indonesia

Science.gov (United States)

Supardan, N.; Panularsih, M.; Darmawan, M.

2018-05-01

Land use change was common in Indonesia, including Bali, especially land conversion from paddy field to other uses. Based on statistics data of Bali 2015, the area of rice fields continues to decline every year, both irrigated and rain-fed rice fields. This study was conducted to analyze land use change and its impact on food security in Denpasar. Land use change dynamics can be observed by multi-temporal land use changes. GIS analysis was performed to obtain spatial land use changes in the study area by using topographic map 1: 25,000 with data sources 2002 and land cover classification of SPOT 6 image acquisition 2015. The results of GIS analysis indicate that settlement and built-up area increased by 1,736 hectares and the area of rice field decreased by 1,695 hectares during 13 years. The increasing of settlement rate was 133,5 ha / year and the decreasing of rice field was 130 ha / year. Meanwhile, the result of food availability analysis shows that the Denpasar City still deficit of rice amount 48,997 tons in 2014, and 69,175 tons in 2015 or only fulfill about 45% of food requirement.

Hot spot in eclipsing dwarf nova IY Ursae Majoris during quiescence and normal outburst

OpenAIRE

Bakowska, K.; Olech, A.

2016-01-01

We present the analysis of hot spot brightness in light curves of the eclipsing dwarf nova IY Ursae Majoris during its normal outburst in March 2013 and in quiescence in April 2012 and in October 2015. Examination of four reconstructed light curves of the hot spot eclipses showed directly that the brightness of the hot spot changed significantly only during the outburst. The brightness of the hot spot, before and after the outburst, was on the same level. Hereby, based on the behaviour of the...

Direct analysis of site-specific N-glycopeptides of serological proteins in dried blood spot samples.

Science.gov (United States)

Choi, Na Young; Hwang, Heeyoun; Ji, Eun Sun; Park, Gun Wook; Lee, Ju Yeon; Lee, Hyun Kyoung; Kim, Jin Young; Yoo, Jong Shin

2017-08-01

Dried blood spot (DBS) samples have a number of advantages, especially with respect to ease of collection, transportation, and storage and to reduce biohazard risk. N-glycosylation is a major post-translational modification of proteins in human blood that is related to a variety of biological functions, including metastasis, cell-cell interactions, inflammation, and immunization. Here, we directly analyzed tryptic N-glycopeptides from glycoproteins in DBS samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS) without centrifugation of blood samples, depletion of major proteins, desalting of tryptic peptides, and enrichment of N-glycopeptides. Using this simple method, we identified a total of 41 site-specific N-glycopeptides from 16 glycoproteins in the DBS samples, from immunoglobulin gamma 1 (IgG-1, 10 mg/mL) down to complement component C7 (50 μg/mL). Of these, 32 N-glycopeptides from 14 glycoproteins were consistently quantified over 180 days stored at room temperature. The major abundant glycoproteins in the DBS samples were IgG-1 and IgG-2, which contain nine asialo-fucosylated complex types of 16 different N-glycopeptide isoforms. Sialo-non-fucosylated complex types were primarily detected in the other glycoproteins such as alpha-1-acid glycoprotein 1, 2, alpha-1-antitypsin, alpha-2-macroglobulin, haptoglobin, hemopexin, Ig alpha 1, 2 chain C region, kininogen-1, prothrombin, and serotransferrin. We first report the characterization of site-specific N-glycoproteins in DBS samples by LC-MS/MS with minimal sample preparation.

Meteorology of Jupiter's Equatorial Hot Spots and Plumes from Cassini

Science.gov (United States)

Choi, David Sanghun; Showman, Adam P.; Vasavada, Ashwin R.; Simon-Miller, Amy A.

2013-01-01

We present an updated analysis of Jupiter's equatorial meteorology from Cassini observations. For two months preceding the spacecraft's closest approach, the Imaging Science Subsystem (ISS) onboard regularly imaged the atmosphere. We created time-lapse movies from this period in order to analyze the dynamics of equatorial hot spots and their interactions with adjacent latitudes. Hot spots are relatively cloud-free regions that emit strongly at 5 lm; improved knowledge of these features is crucial for fully understanding Galileo probe measurements taken during its descent through one. Hot spots are quasistable, rectangular dark areas on visible-wavelength images, with defined eastern edges that sharply contrast with surrounding clouds, but diffuse western edges serving as nebulous boundaries with adjacent equatorial plumes. Hot spots exhibit significant variations in size and shape over timescales of days and weeks. Some of these changes correspond with passing vortex systems from adjacent latitudes interacting with hot spots. Strong anticyclonic gyres present to the south and southeast of the dark areas appear to circulate into hot spots. Impressive, bright white plumes occupy spaces in between hot spots. Compact cirrus-like 'scooter' clouds flow rapidly through the plumes before disappearing within the dark areas. These clouds travel at 150-200 m/s, much faster than the 100 m/s hot spot and plume drift speed. This raises the possibility that the scooter clouds may be more illustrative of the actual jet stream speed at these latitudes. Most previously published zonal wind profiles represent the drift speed of the hot spots at their latitude from pattern matching of the entire longitudinal image strip. If a downward branch of an equatorially-trapped Rossby wave controls the overall appearance of hot spots, however, the westward phase velocity of the wave leads to underestimates of the true jet stream speed.

Roth spots in pernicious anaemia.

Science.gov (United States)

Macauley, Mavin; Nag, Satyajit

2011-04-19

Roth spots are white-centred retinal haemorrhages, previously thought to be pathognomonic for subacute bacterial endocarditis. A number of other conditions can be associated with Roth spots. In this case, the authors describe the association of Roth spots and pernicious anaemia. This association has been rarely described in the medical literature. Correct diagnosis and treatment with intramuscular vitamin B(12) injections resulted in complete resolution of the anaemia and Roth spots. The authors hope to alert clinicians to think of various differentials of Roth spots, and initiate prompt investigation and management.

The effect of defocusing on spot diameter when ablate the silicon surface by femtosecond laser

Science.gov (United States)

Luo, Xinkai; Li, Wei; Wu, Tengfei; Wang, Yu; Zhu, Zhenyu

2018-02-01

Femtosecond laser has been demonstrated to be a prominent tool to manufacture micro scale structure. In the processing, the focusing lens is usually used as the concentrated tool to assemble the original beam to the tiny spot to provide enough energy for ablation. What is more, different focal length means the diverse scale of the focused spot. In common use, various sizes of the spot are required to adjust to the multifarious profiles and substituting the focus lens is the general method. There is no doubt that changing the lens is a fussy job and frequent replacing the lens may cause the lack of stability. In this paper, we report the defocus of the lens to modify the scale of the spot and it is proved to be an effective way to vary the diameter of the focused spot without changing the focus lens.

Comparison of outcomes between overlapping-spot and single-spot photodynamic therapy for circumscribed choroidal hemangioma

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Zhao-An Su

2014-02-01

Full Text Available AIM:To compare the efficacy and safety of photodynamic therapy (PDT with overlapping multiple spots and single spot for treating circumscribed choroidal hemangioma.METHODS:Twenty-two patients (22 eyes with symptomatic circumscribed choroidal hemangioma received PDT treatment. Fourteen patients received overlapping spots (two to three spots PDT, whereas eight patients received single-spot PDT. Laser was used at 50J/cm2 for 83s in the overlapping-spot group and 50J/cm2 for 166s in the single-spot group. Clinical examination, funduscopy, fluorescein angiography, and ultrasonography were performed at baseline and after treatment.RESULTS:The mean follow-up time was 28.5±8.0 months in the overlapping-spot group and 27.0±5.0 months in the single-spot group. Nine patients (64.2% had their vision improved over two lines on the Snellen chart, and five patients showed stable visual acuity in the overlapping-spot group. The mean thickness of tumor decreased from 2.7±0.8mm to 1.2±0.9mm, and the mean greatest tumor linear dimension decreased from 7.4±1.5mm to 4.5±3.5mm after treatment. In the single-spot group, two patients (25% had their vision improved over two lines on the Snellen chart, and six patients had unchanged stable vision. The mean tumor thickness in this group decreased from 2.5±0.7mm to 1.4±1.0mm, and the mean greatest tumor linear dimension decreased from 7.2±1.3mm to 4.7±3.6mm. No significant differences in visual improvement and tumor regression were found between the two groups.CONCLUSION: Overlapping-spot PDT under appropriate treatment parameters and strategies is as effective and safe as single-spot PDT for treating symptomatic circumscribed choroidal hemangioma. Improved or stabilized visual acuity was achieved as a result of tumor regression.

PRDM9 variation strongly influences recombination hot-spot activity and meiotic instability in humans.

Science.gov (United States)

Berg, Ingrid L; Neumann, Rita; Lam, Kwan-Wood G; Sarbajna, Shriparna; Odenthal-Hesse, Linda; May, Celia A; Jeffreys, Alec J

2010-10-01

PRDM9 has recently been identified as a likely trans regulator of meiotic recombination hot spots in humans and mice. PRDM9 contains a zinc finger array that, in humans, can recognize a short sequence motif associated with hot spots, with binding to this motif possibly triggering hot-spot activity via chromatin remodeling. We now report that human genetic variation at the PRDM9 locus has a strong effect on sperm hot-spot activity, even at hot spots lacking the sequence motif. Subtle changes within the zinc finger array can create hot-spot nonactivating or enhancing variants and can even trigger the appearance of a new hot spot, suggesting that PRDM9 is a major global regulator of hot spots in humans. Variation at the PRDM9 locus also influences aspects of genome instability-specifically, a megabase-scale rearrangement underlying two genomic disorders as well as minisatellite instability-implicating PRDM9 as a risk factor for some pathological genome rearrangements.

Two-dimensional gel electrophoresis pattern (pH 6-11) and identification of water-soluble barley seed and malt proteins by mass spectrometry

DEFF Research Database (Denmark)

Bak-Jensen, K.S.; Laugesen, S.; Roepstorff, P.

2004-01-01

A protocol was established for two-dimensional gel electrophoresis (2-DE) of barley seed and malt proteins in the pH range of 6-11. Proteins extracted from flour in a low-salt buffer were focused after cup-loading onto IPG strips. Successful separation in the second dimension was achieved using...... gradient gels in a horizontal SDS-PAGE system. Silver staining of gels visualized around 380 (seed) and 500 (malt) spots. Thirty-seven different proteins from seeds were identified in 60 spots, among these 46 were visualized also in the malt 2-D pattern. Proteins were identified by peptide mass...... in defence against pathogens (21 spots), 4 in storage, folding, and synthesis of proteins, and in nitrogen metabolism (5 spots), 6 in carbohydrate metabolism (11 spots), and 4 in stress and detoxification (9 spots). Six proteins (7 spots) were not grouped in these categories, and 3 were not ascribed...

An Improved 2-Dimensional Gel Electrophoresis Method for Resolving Human Erythrocyte Membrane Proteins.

Science.gov (United States)

Kumar, Manoj; Singh, Rajendra; Meena, Anil; Patidar, Bhagwan S; Prasad, Rajendra; Chhabra, Sunil K; Bansal, Surendra K

2017-01-01

The 2-dimensional gel electrophoresis (2-DE) technique is widely used for the analysis of complex protein mixtures extracted from biological samples. It is one of the most commonly used analytical techniques in proteomics to study qualitative and quantitative protein changes between different states of a cell or an organism (eg, healthy and diseased), conditionally expressed proteins, posttranslational modifications, and so on. The 2-DE technique is used for its unparalleled ability to separate thousands of proteins simultaneously. The resolution of the proteins by 2-DE largely depends on the quality of sample prepared during protein extraction which increases results in terms of reproducibility and minimizes protein modifications that may result in artifactual spots on 2-DE gels. The buffer used for the extraction and solubilization of proteins influences the quality and reproducibility of the resolution of proteins on 2-DE gel. The purification by cleanup kit is another powerful process to prevent horizontal streaking which occurs during isoelectric focusing due to the presence of contaminants such as salts, lipids, nucleic acids, and detergents. Erythrocyte membrane proteins serve as prototypes for multifunctional proteins in various erythroid and nonerythroid cells. In this study, we therefore optimized the selected major conditions of 2-DE for resolving various proteins of human erythrocyte membrane. The modification included the optimization of conditions for sample preparation, cleanup of protein sample, isoelectric focusing, equilibration, and storage of immobilized pH gradient strips, which were further carefully examined to achieve optimum conditions for improving the quality of protein spots on 2-DE gels. The present improved 2-DE analysis method enabled better detection of protein spots with higher quality and reproducibility. Therefore, the conditions established in this study may be used for the 2-DE analysis of erythrocyte membrane proteins for

Significant Change Spotting for Periodic Human Motion Segmentation of Cleaning Tasks Using Wearable Sensors

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Kai-Chun Liu

2017-01-01

Full Text Available The proportion of the aging population is rapidly increasing around the world, which will cause stress on society and healthcare systems. In recent years, advances in technology have created new opportunities for automatic activities of daily living (ADL monitoring to improve the quality of life and provide adequate medical service for the elderly. Such automatic ADL monitoring requires reliable ADL information on a fine-grained level, especially for the status of interaction between body gestures and the environment in the real-world. In this work, we propose a significant change spotting mechanism for periodic human motion segmentation during cleaning task performance. A novel approach is proposed based on the search for a significant change of gestures, which can manage critical technical issues in activity recognition, such as continuous data segmentation, individual variance, and category ambiguity. Three typical machine learning classification algorithms are utilized for the identification of the significant change candidate, including a Support Vector Machine (SVM, k-Nearest Neighbors (kNN, and Naive Bayesian (NB algorithm. Overall, the proposed approach achieves 96.41% in the F1-score by using the SVM classifier. The results show that the proposed approach can fulfill the requirement of fine-grained human motion segmentation for automatic ADL monitoring.

Validation of a prefractionation method followed by two-dimensional electrophoresis – Applied to cerebrospinal fluid proteins from frontotemporal dementia patients

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Sjögren Magnus

2004-11-01

Full Text Available Abstract Background The aim of this study was firstly, to improve and validate a cerebrospinal fluid (CSF prefractionation method followed by two-dimensional electrophoresis (2-DE and secondly, using this strategy to investigate differences between the CSF proteome of frontotemporal dementia (FTD patients and controls. From each subject three ml of CSF was prefractionated using liquid phase isoelectric focusing prior to 2-DE. Results With respect to protein recovery and purification potential, ethanol precipitation of the prefractionated CSF sample was found superior, after testing several sample preparation methods. The reproducibility of prefractionated CSF analyzed on 2-D gels was comparable to direct 2-DE analysis of CSF. The protein spots on the prefractionated 2-D gels had an increased intensity, indicating a higher protein concentration, compared to direct 2-D gels. Prefractionated 2-DE analysis of FTD and control CSF showed that 26 protein spots were changed at least two fold. Using mass spectrometry, 13 of these protein spots were identified, including retinol-binding protein, Zn-α-2-glycoprotein, proapolipoproteinA1, β-2-microglobulin, transthyretin, albumin and alloalbumin. Conclusion The results suggest that the prefractionated 2-DE method can be useful for enrichment of CSF proteins and may provide a new tool to investigate the pathology of neurodegenerative diseases. This study confirmed reduced levels of retinol-binding protein and revealed some new biomarker candidates for FTD.

Characterization of two biologically distinct variants of Tomato spotted wilt virus

Science.gov (United States)

Significant economic losses result on a wide range of crops due to infection with Tomato spotted wilt virus (TSWV). In this study, two TSWV isolates, one from basil and a second from tomato, were established in a common plant host. Viral proteins were monitored over time, plant host ranges were comp...

Spot Accession Protein Protein Unique Secuence Number number ...

Indian Academy of Sciences (India)

36. 702. 2,5. 0,021. 57. 5,26. 143. 13. 33. 865. 2,2. 0,007. 47. 5,24. 321. 6. 20. 492. GRP75_MOUSE. Stress-70 protein, mitochondrial. 2,9. 0,054. 73,4 5,81. 66. 5,18. 118. 6. 10. 663. 2,6. 0,046. 59. 6,18. 162. 10. 33. 717. 2,2. 0,003. 56. 6,2. 259. 11. 43. 1107. 2,1. 0,033. 37. 6,28. 334. 12. 40. 1113. 1,9. 0,013. 36. 6,16. 627. 15.

Sweet Spot Size in Virtual Sound Reproduction: A Temporal Analysis

DEFF Research Database (Denmark)

Lacouture Parodi, Yesenia; Rubak, Per

2009-01-01

The influence of head misalignments on the performance of binaural reproduction systems through loudspeakers is often evaluated in the frequency domain. The changes in magnitude give us an idea of how much of the crosstalk is leaked into the direct signal and therefore a sweet spot performance can......-correlation we estimate the interaural time delay and define a sweet spot. The analysis is based on measurements carried out on 21 different loudspeaker configurations, including two- and four-channels arrangements. Results show that closely spaced loudspeakers are more robust to lateral displacements than wider...... span angles. Additionally, the sweet spot as a function of head rotations increases systematically when the loudspeakers are placed at elevated positions....

Electrophoretic and immunological properties of folate-binding protein isolated from bovine milk

International Nuclear Information System (INIS)

Iwai, Kazuo; Tani, Masako; Fushiki, Tohru

1983-01-01

Changes of the folate-binding protein (FBP) concentration in bovine milk after parturition were investigated. The FBP was highly purified from mature milk by affinity chromatography. The purified FBP showed a single protein band in polyacrylamide gel electrophoresis and was immunologically homogenous in double immunodiffusion. However, in two-dimensional gel electrophoresis, the FBP was separated into several spots in isoelectric focusing in the first dimension, and each spot also showed two molecular weights in SDS-gel electrophoresis in the second dimension. But these FBP molecules were immunologically identical with each other. The neuraminidase treatment obviously diminished the number of isoelectric points of the FBP. Thus, the variety of FBP molecules was at least partially due to the variability of the sialic acid content in the carbohydrate moieties. Moreover, the milk FBP showed species-specificity among mammals immunologically as well as physicochemically. (author)

PmVRP15, a Novel Viral Responsive Protein from the Black Tiger Shrimp, Penaeus monodon, Promoted White Spot Syndrome Virus Replication

Science.gov (United States)

Vatanavicharn, Tipachai; Prapavorarat, Adisak; Jaree, Phattarunda; Somboonwiwat, Kunlaya; Tassanakajon, Anchalee

2014-01-01

Suppression subtractive hybridization of Penaeus monodon hemocytes challenged with white spot syndrome virus (WSSV) has identified the viral responsive gene, PmVRP15, as the highest up-regulated gene ever reported in shrimps. Expression analysis by quantitative real time RT-PCR revealed 9410–fold up-regulated level at 48 h post WSSV injection. Tissue distribution analysis showed that PmVRP15 transcript was mainly expressed in the hemocytes of shrimp. The full-length cDNA of PmVRP15 transcript was obtained and showed no significant similarity to any known gene in the GenBank database. The predicted open reading frame of PmVRP15 encodes for a deduced 137 amino acid protein containing a putative transmembrane helix. Immunofluorescent localization of the PmVRP15 protein revealed it accumulated around the nuclear membrane in all three types of shrimp hemocytes and that the protein was highly up-regulated in WSSV-infected shrimps. Double-stranded RNA interference-mediated gene silencing of PmVRP15 in P. monodon significantly decreased WSSV propagation compared to the control shrimps (injected with GFP dsRNA). The significant decrease in cumulative mortality rate of WSSV-infected shrimp following PmVRP15 knockdown was observed. These results suggest that PmVRP15 is likely to be a nuclear membrane protein and that it acts as a part of WSSV propagation pathway. PMID:24637711

Relation between surface roughness and number of cathode spots of a low-pressure arc

International Nuclear Information System (INIS)

Sato, Atsushi; Iwao, Toru; Yumoto, Motoshige

2008-01-01

A remarkable characteristic of the cathode spot of a low-pressure arc is that it can remove an oxide layer preferentially. Recently, cathode spots of a low-pressure arc have been used for cleaning metal oxide surfaces before thermal spraying or surface modification. Nevertheless, few reports have described the cathode spot movement or the oxide removal process. This experiment was carried out using a Fe+C cathode workpiece and a cylindrical copper anode. The cathode spot movement was recorded using a high-speed video camera. The images were later analysed using plasma image processing. The workpiece surface, which was covered with a 9.67 μm thick oxide, was analysed using laser microscopy after processing. The surface roughness and the number of cathode spots showed no direct relation because the current density per cathode spot did not change according to the number of cathode spots.

SpotCaliper: fast wavelet-based spot detection with accurate size estimation.

Science.gov (United States)

Püspöki, Zsuzsanna; Sage, Daniel; Ward, John Paul; Unser, Michael

2016-04-15

SpotCaliper is a novel wavelet-based image-analysis software providing a fast automatic detection scheme for circular patterns (spots), combined with the precise estimation of their size. It is implemented as an ImageJ plugin with a friendly user interface. The user is allowed to edit the results by modifying the measurements (in a semi-automated way), extract data for further analysis. The fine tuning of the detections includes the possibility of adjusting or removing the original detections, as well as adding further spots. The main advantage of the software is its ability to capture the size of spots in a fast and accurate way. http://bigwww.epfl.ch/algorithms/spotcaliper/ zsuzsanna.puspoki@epfl.ch Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Proteomics analysis of egg white proteins from different egg varieties.

Science.gov (United States)

Wang, Jiapei; Liang, Yue; Omana, Dileep A; Kav, Nat N V; Wu, Jianping

2012-01-11

The market of specialty eggs, such as omega-3-enriched eggs, organic eggs, and free-range eggs, is continuously growing. The nutritional composition of egg yolk can be manipulated by feed diet; however, it is not known if there is any difference in the composition of egg white proteins among different egg varieties. The purpose of the study was to compare the egg white proteins among six different egg varieties using proteomics analysis. Egg white proteins were analyzed using two-dimensional gel electrophoresis (2-DE), and 89 protein spots were subjected to LC-MS/MS. A total of 23 proteins, belonging to Gallus gallus , were identified from 72 detected protein spots. A quiescence-specific protein precursor in egg white was identified for the first time in this study. Significant differences in the abundant levels of 19 proteins (from 65 protein spots) were observed among six egg varieties. Four proteins, ovalbumin-related protein Y, cystatin, avidin, and albumin precursor, were not different among these six egg varieties. These findings suggest that the abundance, but not the composition, of egg white proteins varied among the egg varieties.

Measurement and application of purine derivatives: Creatinine ratio in spot urine samples of ruminants

International Nuclear Information System (INIS)

Chen, X.B.; Jayasuriya, M.C.N.; Makkar, H.P.S.

2004-01-01

The daily excretion of purine derivatives in urine has been used to estimate the supply of microbial protein to ruminant animals. The method provides a simple and non-invasive tool to indicate the nutritional status of farm animals. However due to the need for complete collection of urine the potential application at farm level is restricted. Research conducted under the FAO/IAEA Co-ordinated Research Project has indicated that it is possible to use the purine derivatives:creatinine ratio measured in several spot urine samples collected within a day, as an index of microbial protein supply in a banding system for farm application. Some theoretical and experimental aspects in the measurement of purine derivatives:creatinine ratio in spot urine samples and the possible application of the banding system at the farm level are discussed. (author)

Exogenous Expressions of FTO Wild-Type and R316Q Mutant Proteins Caused an Increase in HNRPK Levels in 3T3-L1 Cells as Demonstrated by DIGE Analysis

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Nil Guzel

2017-01-01

Full Text Available Fat mass and obesity-associated protein is an enzyme that oxidatively demethylates DNA. Although there are numerous studies regarding the catalytic function of FTO, the overall existence or absence of FTO on cellular proteome has not been investigated. This study investigated the changes in the soluble proteome of 3T3-L1 cells upon expression of the WT and the mutant (R316Q FTO proteins. Protein extracts prepared from 3T3-L1 cells expressing either the WT or the mutant FTO proteins were used in DIGE experiments. Analysis of the data revealed the number of spots matched to every member and there were 350 ± 20 spots with 30.5% overall mean coefficient of variation. Eleven regulated protein spots were excised from the gels and identified by MALDI-TOF/TOF. One of the identified proteins was heterogeneous nuclear ribonucleoprotein K, which displayed more than 2.6- and 3.7-fold increases in its abundance in the WT and the mutant FTO expressing cells, respectively. Western blot analysis validated these observations. This is the first study revealing the presence of a parallel increase in expressions of FTO and HNRNPK proteins. This increase may codictate the metabolic changes occurring in the cell and may attribute a significance to HNRNPK in FTO-associated transformations.

Roth spots in pernicious anaemia

OpenAIRE

Macauley, Mavin; Nag, Satyajit

2011-01-01

Roth spots are white-centred retinal haemorrhages, previously thought to be pathognomonic for subacute bacterial endocarditis. A number of other conditions can be associated with Roth spots. In this case, the authors describe the association of Roth spots and pernicious anaemia. This association has been rarely described in the medical literature. Correct diagnosis and treatment with intramuscular vitamin B12 injections resulted in complete resolution of the anaemia and Roth spots. The author...

Ionospheric hot spot at high latitudes

International Nuclear Information System (INIS)

Schunk, R.W.; Sojka, J.J.

1982-01-01

A hot spot (or spots) can occur in the high-latitude ionosphere depending on the plasma convection pattern. The hot spot corresponds to a small magnetic local time-magnetic latitude region of elevated ion temperatures located near the dusk and/or dawn meridians. For asymmetric convection electric field patterns, with enhanced flow in either the dusk or dawn sector of the polar cap, a single hot spot should occur in association with the strong convection cell. However, on geomagnetically disturbed days, two strong convection cells can occur, and hence, two hot spots should exist. The hot spot should be detectable when the electric field in the strong convection cell exceeds about 40 mV m -1 . For electric fields of the order of 100 mV m -1 in the convection cell, the ion temperature in the hot spot is greatest at low altitudes, reaching 4000 0 K at 160 km, and decreases with altitude in the F-region. An ionospheric hot spot (or spots) can be expected at all seasons and for a wide range of solar cycle conditions

Predicting "Hot" and "Warm" Spots for Fragment Binding.

Science.gov (United States)

Rathi, Prakash Chandra; Ludlow, R Frederick; Hall, Richard J; Murray, Christopher W; Mortenson, Paul N; Verdonk, Marcel L

2017-05-11

Computational fragment mapping methods aim to predict hotspots on protein surfaces where small fragments will bind. Such methods are popular for druggability assessment as well as structure-based design. However, to date researchers developing or using such tools have had no clear way of assessing the performance of these methods. Here, we introduce the first diverse, high quality validation set for computational fragment mapping. The set contains 52 diverse examples of fragment binding "hot" and "warm" spots from the Protein Data Bank (PDB). Additionally, we describe PLImap, a novel protocol for fragment mapping based on the Protein-Ligand Informatics force field (PLIff). We evaluate PLImap against the new fragment mapping test set, and compare its performance to that of simple shape-based algorithms and fragment docking using GOLD. PLImap is made publicly available from https://bitbucket.org/AstexUK/pli .

A virus-binding hot spot on human angiotensin-converting enzyme 2 is critical for binding of two different coronaviruses.

Science.gov (United States)

Wu, Kailang; Chen, Lang; Peng, Guiqing; Zhou, Wenbo; Pennell, Christopher A; Mansky, Louis M; Geraghty, Robert J; Li, Fang

2011-06-01

How viruses evolve to select their receptor proteins for host cell entry is puzzling. We recently determined the crystal structures of NL63 coronavirus (NL63-CoV) and SARS coronavirus (SARS-CoV) receptor-binding domains (RBDs), each complexed with their common receptor, human angiotensin-converting enzyme 2 (hACE2), and proposed the existence of a virus-binding hot spot on hACE2. Here we investigated the function of this hypothetical hot spot using structure-guided biochemical and functional assays. The hot spot consists of a salt bridge surrounded by hydrophobic tunnel walls. Mutations that disturb the hot spot structure have significant effects on virus/receptor interactions, revealing critical energy contributions from the hot spot structure. The tunnel structure at the NL63-CoV/hACE2 interface is more compact than that at the SARS-CoV/hACE2 interface, and hence RBD/hACE2 binding affinities are decreased either by NL63-CoV mutations decreasing the tunnel space or by SARS-CoV mutations increasing the tunnel space. Furthermore, NL63-CoV RBD inhibits hACE2-dependent transduction by SARS-CoV spike protein, a successful application of the hot spot theory that has the potential to become a new antiviral strategy against SARS-CoV infections. These results suggest that the structural features of the hot spot on hACE2 were among the driving forces for the convergent evolution of NL63-CoV and SARS-CoV.

Variable-spot ion beam figuring

International Nuclear Information System (INIS)

Wu, Lixiang; Qiu, Keqiang; Fu, Shaojun

2016-01-01

This paper introduces a new scheme of ion beam figuring (IBF), or rather variable-spot IBF, which is conducted at a constant scanning velocity with variable-spot ion beam collimated by a variable diaphragm. It aims at improving the reachability and adaptation of the figuring process within the limits of machine dynamics by varying the ion beam spot size instead of the scanning velocity. In contrast to the dwell time algorithm in the conventional IBF, the variable-spot IBF adopts a new algorithm, which consists of the scan path programming and the trajectory optimization using pattern search. In this algorithm, instead of the dwell time, a new concept, integral etching time, is proposed to interpret the process of variable-spot IBF. We conducted simulations to verify its feasibility and practicality. The simulation results indicate the variable-spot IBF is a promising alternative to the conventional approach.

[Study on Hexagonal Super-Lattice Pattern with Light Spot and Dim Spot in Dielectric Barrier Discharge by Optical Emission Spectra].

Science.gov (United States)

Liu, Ying; Dong, Li-fang; Niu, Xue-jiao; Zhang, Chao

2016-02-01

The hexagonal super-lattice pattern composed of the light spot and the dim spot is firstly observed and investigated in the discharge of gas mixture of air and argon by using the dielectric barrier discharge device with double water electrodes. It is found that the dim spot is located at the center of its surrounding three light spots by observing the discharge image. Obviously, the brightness of the light spot and the dim spot are different, which indicates that the plasma states of the light spot and the dim spot may be different. The optical emission spectrum method is used to further study the several plasma parameters of the light spot and the dim spot in different argon content. The emission spectra of the N₂ second positive band (C³IIu --> B³IIg) are measured, from which the molecule vibration temperatures of the light spot and the dim spot are calculated. Based on the relative intensity ratio of the line at 391.4 nm and the N₂ line at 394.1 nm, the average electron energies of the light spot and the dim spot are investigated. The broadening of spectral line 696.57 nm (2P₂-1S₅) is used to study the electron densities of the light spot and the dim spot. The experiment shows that the molecule vibration temperature, average electron energy and the electron density of the dim spot are higher than those of the light spot in the same argon content. The molecule vibration temperature and electron density of the light spot and dim spot increase with the argon content increasing from 70% to 95%, while average electron energies of the light spot and dim spot decrease gradually. The short-exposure image recorded by a high speed video camera shows that the dim spot results from the surface discharges (SDs). The surface discharge induced by the volume discharge (VD) has the decisive effect on the formation of the dim spot. The experiment above plays an important role in studying the formation mechanism of the hexagonal super-lattice pattern with light spot and

Changes in protein patterns and in vivo protein synthesis during senescence of hibiscus petals

International Nuclear Information System (INIS)

Woodson, W.R.; Handa, A.K.

1986-01-01

Changes in proteins associated with senescence of the flowers of Hibiscus rosa-sinensis was studied using SDS-PAGE. Total extractable protein from petals decreased with senescence. Changes were noted in patterns of proteins from aging petals. Flower opening and senescence was associated with appearance and disappearance of several polypeptides. One new polypeptide with an apparent mw of 41 kd was first seen the day of flower opening and increased to over 9% of the total protein content of senescent petal tissue. Protein synthesis during aging was investigated by following uptake and incorporation of 3 H-leucine into TCA-insoluble fraction of petal discs. Protein synthesis, as evidenced by the percent of label incorporated into the TCA-insoluble fraction, was greatest (32%) the day before flower opening. Senescent petal tissue incorporated 4% of label taken up into protein. Proteins were separated by SDS-PAGE and labelled polypeptides identified by fluorography. In presenescent petal tissue, radioactivity was distributed among several major polypeptides. In senescent tissue, much of the radioactivity was concentrated in the 41 kd polypeptide

Numerical optimisation in spot detector design

NARCIS (Netherlands)

van der Heijden, Ferdinand; Apperloo, W.; Spreeuwers, Lieuwe Jan

1997-01-01

Spots are image details resulting from objects, the projections of which are so small that the inner structure of these objects cannot be resolved from their image. Spot detectors are image operators aiming at the detection and localisation of spots in the image. Most spot detectors can be tuned

The complete mitochondrial genome of the three-spot seahorse, Hippocampus trimaculatus (Teleostei, Syngnathidae).

Science.gov (United States)

Chang, Chia-Hao; Shao, Kwang-Tsao; Lin, Yeong-Shin; Liao, Yun-Chih

2013-12-01

The complete mitochondrial genome of the three-spot seahorse was sequenced using a polymerase chain reaction-based method. The total length of mitochondrial DNA is 16,535 bp and includes 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and a control region. The mitochondrial gene order of the three-spot seahorse also conforms to the distinctive vertebrate mitochondrial gene order. The base composition of the genome is A (32.7%), T (29.3%), C (23.4%), and G (14.6%) with an A + T-rich hallmark as that of other vertebrate mitochondrial genomes.

9 CFR 149.4 - Spot audit.

Science.gov (United States)

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Spot audit. 149.4 Section 149.4... LIVESTOCK IMPROVEMENT VOLUNTARY TRICHINAE CERTIFICATION PROGRAM § 149.4 Spot audit. (a) In addition to regularly scheduled site audits, certified production sites will be subject to spot audits. (1) Random spot...

Exclusion of EDNRB and KIT as the basis for white spotting in Border Collies.

Science.gov (United States)

Metallinos, D; Rine, J

2000-01-01

White spotting patterns in mammals can be caused by mutations in the genes for the endothelin B receptor and c-Kit, whose protein products are necessary for proper migration, differentiation or survival of the melanoblast population of cells. Although there are many different dog breeds that segregate white spotting patterns, no genes have been identified that are linked to these phenotypes. An intercross was generated from a female Newfoundland and a male Border Collie and the white spotting phenotypes of the intercross progeny were evaluated by measuring percentage surface area of white in the puppies. The Border Collie markings segregated as a simple autosomal recessive (7/25 intercross progeny had the phenotype). Two candidate genes, for the endothelin B receptor (EDNRB) and c-Kit (KIT), were evaluated for segregation with the white spotting pattern. Polymorphisms between the Border Collie and Newfoundland were identified for EDNRB using Southern analysis after a portion of the canine gene had been cloned. Polymorphisms for KIT were identified using a microsatellite developed from a bacterial artificial chromosome containing the canine gene. Both EDNRB and KIT were excluded as a cause of the white spotting pattern in at least two of the intercross progeny. Although these genes have been implicated in white spotting in other mammals, including horses, pigs, cows, mice and rats, they do not appear to be responsible for the white spotting pattern found in the Border Collie breed of dog.

Yeast Surface Display of Two Proteins Previously Shown to Be Protective Against White Spot Syndrome Virus (WSSV) in Shrimp.

Science.gov (United States)

Ananphongmanee, Vorawit; Srisala, Jiraporn; Sritunyalucksana, Kallaya; Boonchird, Chuenchit

2015-01-01

Cell surface display using the yeasts Saccharomyces cerevisiae and Pichia pastoris has been extensively developed for application in bioindustrial processes. Due to the rigid structure of their cell walls, a number of proteins have been successfully displayed on their cell surfaces. It was previously reported that the viral binding protein Rab7 from the giant tiger shrimp Penaeus monodon (PmRab7) and its binding partner envelope protein VP28 of white spot syndrome virus (WSSV) could independently protect shrimp against WSSV infection. Thus, we aimed to display these two proteins independently on the cell surfaces of 2 yeast clones with the ultimate goal of using a mixture of the two clones as an orally deliverable, antiviral agent to protect shrimp against WSSV infection. PmRab7 and VP28 were modified by N-terminal tagging to the C-terminal half of S. cerevisiae α-agglutinin. DNA fragments, harboring fused-gene expression cassettes under control of an alcohol oxidase I (AOX1) promoter were constructed and used to transform the yeast cells. Immunofluorescence microscopy with antibodies specific to both proteins demonstrated that mutated PmRab7 (mPmRab7) and partial VP28 (pVP28) were localized on the cell surfaces of the respective clones, and fluorescence intensity for each was significantly higher than that of control cells by flow cytometry. Enzyme-linked immunosorbant assay (ELISA) using cells displaying mPmRab7 or pVP28 revealed that the binding of specific antibodies for each was dose-dependent, and could be saturated. In addition, the binding of mPmRab7-expressing cells with free VP28, and vice versa was dose dependent. Binding between the two surface-expressed proteins was confirmed by an assay showing agglutination between cells expressing complementary mPmRab7 and pVP28. In summary, our genetically engineered P. pastoris can display biologically active mPmRab7 and pVP28 and is now ready for evaluation of efficacy in protecting shrimp against WSSV by oral

Yeast Surface Display of Two Proteins Previously Shown to Be Protective Against White Spot Syndrome Virus (WSSV in Shrimp.

Directory of Open Access Journals (Sweden)

Vorawit Ananphongmanee

Full Text Available Cell surface display using the yeasts Saccharomyces cerevisiae and Pichia pastoris has been extensively developed for application in bioindustrial processes. Due to the rigid structure of their cell walls, a number of proteins have been successfully displayed on their cell surfaces. It was previously reported that the viral binding protein Rab7 from the giant tiger shrimp Penaeus monodon (PmRab7 and its binding partner envelope protein VP28 of white spot syndrome virus (WSSV could independently protect shrimp against WSSV infection. Thus, we aimed to display these two proteins independently on the cell surfaces of 2 yeast clones with the ultimate goal of using a mixture of the two clones as an orally deliverable, antiviral agent to protect shrimp against WSSV infection. PmRab7 and VP28 were modified by N-terminal tagging to the C-terminal half of S. cerevisiae α-agglutinin. DNA fragments, harboring fused-gene expression cassettes under control of an alcohol oxidase I (AOX1 promoter were constructed and used to transform the yeast cells. Immunofluorescence microscopy with antibodies specific to both proteins demonstrated that mutated PmRab7 (mPmRab7 and partial VP28 (pVP28 were localized on the cell surfaces of the respective clones, and fluorescence intensity for each was significantly higher than that of control cells by flow cytometry. Enzyme-linked immunosorbant assay (ELISA using cells displaying mPmRab7 or pVP28 revealed that the binding of specific antibodies for each was dose-dependent, and could be saturated. In addition, the binding of mPmRab7-expressing cells with free VP28, and vice versa was dose dependent. Binding between the two surface-expressed proteins was confirmed by an assay showing agglutination between cells expressing complementary mPmRab7 and pVP28. In summary, our genetically engineered P. pastoris can display biologically active mPmRab7 and pVP28 and is now ready for evaluation of efficacy in protecting shrimp against

Spot formation of radiation particles by electrochemical etching

International Nuclear Information System (INIS)

Nozaki, Tetsuya

1999-01-01

An electrochemical etching (ECE) spot formation from the top of chemical etching (CE) spot was confirmed by a series of experiments. One of polycarbonate (Iupilon) could not make the spot, because ECE spot had grown up before the microscope confirming the CE spot. Clear CEC spots by α-ray and neutron were found on Harzlas and Baryotrak, both improvements of CR-39. Under the same etching conditions, the growth of ECE spot on Harzlas was more rapid than Baryotrak, but both spots were almost the same. All CE spot by α-ray produced the CEC spots, but a part of CE circle spot by neutron formed them. (S.Y.)

Shrimp hemocyte homeostasis-associated protein (PmHHAP) interacts with WSSV134 to control apoptosis in white spot syndrome virus infection.

Science.gov (United States)

Apitanyasai, Kantamas; Amparyup, Piti; Charoensapsri, Walaiporn; Sangsuriya, Pakkakul; Tassanakajon, Anchalee

2018-05-01

Hemocyte homeostasis-associated protein (PmHHAP) was first identified as a viral-responsive gene, due to a high upregulation in transcription following white spot syndrome virus (WSSV) infection. Functional studies using RNA interference have suggested that PmHHAP is involved in hemocyte homeostasis by controlling apoptosis during WSSV infection. In this study, the role of PmHHAP in host-viral interactions was further investigated. Yeast two-hybrid assay and co-immunoprecipitation revealed that PmHHAP binds to an anti-apoptosis protein, WSSV134. The viral protein WSSV134 is a late protein of WSSV, expressed 24 h post infection (hpi). Gene silencing of WSSV134 in WSSV-infected shrimp resulted in a reduction of the expression level of the viral replication marker genes VP28, wsv477, and ie-1, which suggests that WSSV134 is likely involved in viral propagation. However, co-silencing of PmHHAP and WSSV134 counteracted the effects on WSSV infection, which implies the importance of the host-pathogen interaction between PmHHAP and WSSV134 in WSSV infection. In addition, caspase 3/7 activity was noticeably induced in the PmHHAP and WSSV134 co-silenced shrimp upon WSSV infection. Moreover, PmHHAP and WSSV134 inhibited caspase-induced activation of PmCasp in vitro in a non-competitive manner. Taken together, these results suggest that PmHHAP and WSSV134 play a role in the host-pathogen interaction and work concordantly to control apoptosis in WSSV infection. Copyright © 2018 Elsevier Ltd. All rights reserved.

Differential antigenic protein recovery from Taenia solium cyst tissues using several detergents.

Science.gov (United States)

Navarrete-Perea, José; Orozco-Ramírez, Rodrigo; Moguel, Bárbara; Sciutto, Edda; Bobes, Raúl J; Laclette, Juan P

2015-07-01

Human and porcine cysticercosis is caused by the larval stage of the flatworm Taenia solium (Cestoda). The protein extracts of T. solium cysts are complex mixtures including cyst's and host proteins. Little is known about the influence of using different detergents in the efficiency of solubilization-extraction of these proteins, including relevant antigens. Here, we describe the use of CHAPS, ASB-14 and Triton X-100, alone or in combination in the extraction buffers, as a strategy to notably increase the recovery of proteins that are usually left aside in insoluble fractions of cysts. Using buffer with CHAPS alone, 315 protein spots were detected through 2D-PAGE. A total of 255 and 258 spots were detected using buffers with Triton X-100 or ASB-14, respectively. More protein spots were detected when detergents were combined, i.e., 2% CHAPS, 1% Triton X-100 and 1% ASB-14 allowed detection of up to 368 spots. Our results indicated that insoluble fractions of T. solium cysts were rich in antigens, including several glycoproteins that were sensitive to metaperiodate treatment. Host proteins, a common component in protein extracts of cysts, were present in larger amounts in soluble than insoluble fractions of cysts proteins. Finally, antigens present in the insoluble fraction were more appropriate as a source of antigens for diagnostic procedures. Copyright © 2015 Elsevier B.V. All rights reserved.

Proteomic Profiling Comparing the Effects of Different Heat Treatments on Camel (Camelus dromedarius) Milk Whey Proteins.

Science.gov (United States)

Benabdelkamel, Hicham; Masood, Afshan; Alanazi, Ibrahim O; Alzahrani, Dunia A; Alrabiah, Deema K; AlYahya, Sami A; Alfadda, Assim A

2017-03-28

Camel milk is consumed in the Middle East because of its high nutritional value. Traditional heating methods and the duration of heating affect the protein content and nutritional quality of the milk. We examined the denaturation of whey proteins in camel milk by assessing the effects of temperature on the whey protein profile at room temperature (RT), moderate heating at 63 °C, and at 98 °C, for 1 h. The qualitative and quantitative variations in the whey proteins before and after heat treatments were determined using quantitative 2D-difference in gel electrophoresis (DIGE)-mass spectrometry. Qualitative gel image analysis revealed a similar spot distribution between samples at RT and those heated at 63 °C, while the spot distribution between RT and samples heated at 98 °C differed. One hundred sixteen protein spots were determined to be significantly different ( p protein spots were decreased in common in both the heat-treated samples and an additional 25 spots were further decreased in the 98 °C sample. The proteins with decreased abundance included serum albumin, lactadherin, fibrinogen β and γ chain, lactotransferrin, active receptor type-2A, arginase-1, glutathione peroxidase-1 and, thiopurine S, etc. Eight protein spots were increased in common to both the samples when compared to RT and included α-lactalbumin, a glycosylation-dependent cell adhesion molecule. Whey proteins present in camel milk were less affected by heating at 63 °C than at 98 °C. This experimental study showed that denaturation increased significantly as the temperature increased from 63 to 98 °C.

Measurements of three dimensional residual stress distribution on laser irradiated spot

International Nuclear Information System (INIS)

Tanaka, Hirotomo; Akita, Koichi; Ohya, Shin-ichi; Sano, Yuji; Naito, Hideki

2004-01-01

Three dimensional residual stress distributions on laser irradiated spots were measured using synchrotron radiation to study the basic mechanism of laser peening. A water-immersed sample of high tensile strength steel was irradiated with Q-switched and frequency-doubled Nd:YAG laser. The residual stress depth profile of the sample was obtained by alternately repeating the measurement and surface layer removal by electrolytic polishing. Tensile residual stresses were observed on the surface of all irradiated spots, whereas residual stress changed to compressive just beneath the surface. The depth of compressive residual stress imparted by laser irradiation and plastic deformation zone increased with increasing the number of laser pulses irradiated on the same spot. (author)

Association of poly-purine/poly-pyrimidine sequences with meiotic recombination hot spots

Directory of Open Access Journals (Sweden)

Pitt Joel PW

2006-07-01

Full Text Available Abstract Background Meiotic recombination events have been found to concentrate in 1–2.5 kilo base regions, but these recombination hot spots do not share a consensus sequence and why they occur at specific sites is not fully understood. Some previous evidence suggests that poly-purine/poly-pyrimidine (poly-pu/py tracts (PPTs, a class of sequence with distinctive biochemical properties, could be involved in recombination, but no general association of PPTs with meiotic recombination hot spots has previously been reported. Results We used computational methods to investigate in detail the relationship between PPTs and hot spots. We show statistical associations of PPT frequency with hot spots of meiotic recombination initiating lesions, double-strand breaks, in the genome of the yeast S. cerevisiae and with experimentally well characterized human meiotic recombination hot spots. Supporting a possible role of poly-pu/py-rich sequences in hot spot recombination, we also found that all three single nucleotide polymorphisms previously shown to be associated with human hot spot activity changes occur within sequence contexts of 14 bp or longer that are 85% or more poly-pu/py and at least 70% G/C. These polymorphisms are all close to the hot spot mid points. Comparing the sequences of experimentally characterized human hot spots with the orthologous regions of the chimpanzee genome previously shown not to contain hot spots, we found that in all five cases in which comparisons for the hot spot central regions are possible with publicly available sequence data, there are differences near the human hot spot mid points within sequences 14 bp or longer consisting of more than 80% poly-pu/py and at least 50% G/C. Conclusion Our results, along with previous evidence for the unique biochemical properties and recombination-stimulating potential of poly-pu/py-rich sequences, suggest that the possible functional involvement of this type of sequence in meiotic

Poisson Spot with Magnetic Levitation

Science.gov (United States)

Hoover, Matthew; Everhart, Michael; D'Arruda, Jose

2010-01-01

In this paper we describe a unique method for obtaining the famous Poisson spot without adding obstacles to the light path, which could interfere with the effect. A Poisson spot is the interference effect from parallel rays of light diffracting around a solid spherical object, creating a bright spot in the center of the shadow.

Substantial conformational change mediated by charge-triad residues of the death effector domain in protein-protein interactions.

Directory of Open Access Journals (Sweden)

Edward C Twomey

Full Text Available Protein conformational changes are commonly associated with the formation of protein complexes. The non-catalytic death effector domains (DEDs mediate protein-protein interactions in a variety of cellular processes, including apoptosis, proliferation and migration, and glucose metabolism. Here, using NMR residual dipolar coupling (RDC data, we report a conformational change in the DED of the phosphoprotein enriched in astrocytes, 15 kDa (PEA-15 protein in the complex with a mitogen-activated protein (MAP kinase, extracellular regulated kinase 2 (ERK2, which is essential in regulating ERK2 cellular distribution and function in cell proliferation and migration. The most significant conformational change in PEA-15 happens at helices α2, α3, and α4, which also possess the highest flexibility among the six-helix bundle of the DED. This crucial conformational change is modulated by the D/E-RxDL charge-triad motif, one of the prominent structural features of DEDs, together with a number of other electrostatic and hydrogen bonding interactions on the protein surface. Charge-triad motif promotes the optimal orientation of key residues and expands the binding interface to accommodate protein-protein interactions. However, the charge-triad residues are not directly involved in the binding interface between PEA-15 and ERK2.

New 40Ar/39Ar age progression for the Louisville hot spot trail and implications for inter-hot spot motion

Science.gov (United States)

Koppers, Anthony A. P.; Gowen, Molly D.; Colwell, Lauren E.; Gee, Jeffrey S.; Lonsdale, Peter F.; Mahoney, John J.; Duncan, Robert A.

2011-12-01

hot spot motions, therefore provide a better fit to the new Louisville age data. The fit is particularly good for seamounts younger than 50 Ma, a period for which there is little predicted motion for the Louisville hot spot and little inter-hot spot motion with Hawaii. However, discrepancies in the Louisville age-distance record prior to 50 Ma indicate there is an extra source of inter-hot spot motion between Louisville and the other Pacific hot spots that was not corrected for in the global S04G model. Finally, based on six new 40Ar/39Ar age dates, the 169°W bend in the Louisville seamount trail seems to have formed at least 3 Myr before the formation of the Hawaiian-Emperor bend. The timing of the most acute parts of both bends thus appears to be asynchronous, which would require other processes (e.g., plume motions) than a global plate motion change between 50 and 47 Ma to explain these two observations.

Immunoproteomic analysis of proteins from unsporulated Oocysts of Eimeria tenella in MALDI TOF/TOF tandem mass spectrometry

Science.gov (United States)

Immunoproteomic approaches were conducted to identify antigenic proteins from the total proteins of unsporulated oocysts of Eimeria tenella (E. tenella). Approximately 101 protein spots were recognized by chicken sera infected experimentally with E. tenella. Fourty-six spots of unsporulated oocysts ...

Statistical hot spot analysis of reactor cores

International Nuclear Information System (INIS)

Schaefer, H.

1974-05-01

This report is an introduction into statistical hot spot analysis. After the definition of the term 'hot spot' a statistical analysis is outlined. The mathematical method is presented, especially the formula concerning the probability of no hot spots in a reactor core is evaluated. A discussion with the boundary conditions of a statistical hot spot analysis is given (technological limits, nominal situation, uncertainties). The application of the hot spot analysis to the linear power of pellets and the temperature rise in cooling channels is demonstrated with respect to the test zone of KNK II. Basic values, such as probability of no hot spots, hot spot potential, expected hot spot diagram and cumulative distribution function of hot spots, are discussed. It is shown, that the risk of hot channels can be dispersed equally over all subassemblies by an adequate choice of the nominal temperature distribution in the core

Protein Structural Change Data - PSCDB | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available List Contact us PSCDB Protein Structural Change Data Data detail Data name Protein Structural Change Data DO...History of This Database Site Policy | Contact Us Protein Structural Change Data - PSCDB | LSDB Archive ...

40 CFR 86.163-00 - Spot check correlation procedures for vehicles tested using a simulation of the environmental...

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 18 2010-07-01 2010-07-01 false Spot check correlation procedures for... Complete Heavy-Duty Vehicles; Test Procedures § 86.163-00 Spot check correlation procedures for vehicles... running change approval, each model year for any manufacturer undergoing the spot checking procedures of...

Hot Spot Removal System: System description

Energy Technology Data Exchange (ETDEWEB)

NONE

1997-09-01

Hazardous wastes contaminated with radionuclides, chemicals, and explosives exist across the Department of Energy complex and need to be remediated due to environmental concerns. Currently, an opportunity is being developed to dramatically reduce remediation costs and to assist in the acceleration of schedules associated with these wastes by deploying a Hot Spot Removal System. Removing the hot spot from the waste site will remove risk driver(s) and enable another, more cost effective process/option/remedial alternative (i.e., capping) to be applied to the remainder of the site. The Hot Spot Removal System consists of a suite of technologies that will be utilized to locate and remove source terms. Components of the system can also be used in a variety of other cleanup activities. This Hot Spot Removal System Description document presents technologies that were considered for possible inclusion in the Hot Spot Removal System, technologies made available to the Hot Spot Removal System, industrial interest in the Hot Spot Removal System`s subsystems, the schedule required for the Hot Spot Removal System, the evaluation of the relevant technologies, and the recommendations for equipment and technologies as stated in the Plan section.

Hot Spot Removal System: System description

International Nuclear Information System (INIS)

1997-09-01

Hazardous wastes contaminated with radionuclides, chemicals, and explosives exist across the Department of Energy complex and need to be remediated due to environmental concerns. Currently, an opportunity is being developed to dramatically reduce remediation costs and to assist in the acceleration of schedules associated with these wastes by deploying a Hot Spot Removal System. Removing the hot spot from the waste site will remove risk driver(s) and enable another, more cost effective process/option/remedial alternative (i.e., capping) to be applied to the remainder of the site. The Hot Spot Removal System consists of a suite of technologies that will be utilized to locate and remove source terms. Components of the system can also be used in a variety of other cleanup activities. This Hot Spot Removal System Description document presents technologies that were considered for possible inclusion in the Hot Spot Removal System, technologies made available to the Hot Spot Removal System, industrial interest in the Hot Spot Removal System''s subsystems, the schedule required for the Hot Spot Removal System, the evaluation of the relevant technologies, and the recommendations for equipment and technologies as stated in the Plan section

Training-induced changes in membrane transport proteins of human skeletal muscle

DEFF Research Database (Denmark)

Juel, C.

2006-01-01

Training improves human physical performance by inducing structural and cardiovascular changes, metabolic changes, and changes in the density of membrane transport proteins. This review focuses on the training-induced changes in proteins involved in sarcolemmal membrane transport. It is concluded...

Isotopic Changes During Digestion: Protein

Science.gov (United States)

Tuross, N.

2013-12-01

Nutrient and hydrological inputs traverse a complicated route of pH, enzymatic and cellular processes in digestion in higher animals. The end products of digestion are the starting products for biosynthesis that are often used to interpret past life-ways. Using an artificial gut system, the isotopic changes (dD, d18O, d13C and d15N) of protein are documented. Three separate protein sources are subjected to the conditions, chemical and enzymatic, found in the stomach and upper small intestine with only a small shift in the oxygen isotopic composition of the proteins observed. Middle to lower small intestine parameters produced both greater isotopic effects and significantly lower molecular weight products. The role of the gastric enterocyte and the likely involvement of the internal milieu of this cell in the isotopic composition of amino acids that are transported to the liver are reported.

Methodology and software to detect viral integration site hot-spots

Science.gov (United States)

2011-01-01

Background Modern gene therapy methods have limited control over where a therapeutic viral vector inserts into the host genome. Vector integration can activate local gene expression, which can cause cancer if the vector inserts near an oncogene. Viral integration hot-spots or 'common insertion sites' (CIS) are scrutinized to evaluate and predict patient safety. CIS are typically defined by a minimum density of insertions (such as 2-4 within a 30-100 kb region), which unfortunately depends on the total number of observed VIS. This is problematic for comparing hot-spot distributions across data sets and patients, where the VIS numbers may vary. Results We develop two new methods for defining hot-spots that are relatively independent of data set size. Both methods operate on distributions of VIS across consecutive 1 Mb 'bins' of the genome. The first method 'z-threshold' tallies the number of VIS per bin, converts these counts to z-scores, and applies a threshold to define high density bins. The second method 'BCP' applies a Bayesian change-point model to the z-scores to define hot-spots. The novel hot-spot methods are compared with a conventional CIS method using simulated data sets and data sets from five published human studies, including the X-linked ALD (adrenoleukodystrophy), CGD (chronic granulomatous disease) and SCID-X1 (X-linked severe combined immunodeficiency) trials. The BCP analysis of the human X-linked ALD data for two patients separately (774 and 1627 VIS) and combined (2401 VIS) resulted in 5-6 hot-spots covering 0.17-0.251% of the genome and containing 5.56-7.74% of the total VIS. In comparison, the CIS analysis resulted in 12-110 hot-spots covering 0.018-0.246% of the genome and containing 5.81-22.7% of the VIS, corresponding to a greater number of hot-spots as the data set size increased. Our hot-spot methods enable one to evaluate the extent of VIS clustering, and formally compare data sets in terms of hot-spot overlap. Finally, we show that the

ESA uncovers Geminga's `hot spot'

Science.gov (United States)

2004-07-01

16 July 2004 Astronomers using ESA’s X-ray observatory XMM-Newton have detected a small, bright ‘hot spot’ on the surface of the neutron star called Geminga, 500 light-years away. The hot spot is the size of a football field and is caused by the same mechanism producing Geminga’s X-ray tails. This discovery identifies the missing link between the X-ray and gamma-ray emission from Geminga. hi-res Size hi-res: 1284 kb Credits: ESA, P. Caraveo (IASF, Milan) Geminga's hot spot This figure shows the effects of charged particles accelerated in the magnetosphere of Geminga. Panel (a) shows an image taken with the EPIC instrument on board the XMM-Newton observatory. The bright tails, made of particles kicked out by Geminga’s strong magnetic field, trail the neutron star as it moves about in space. Panel (b) shows how electrically charged particles interact with Geminga’s magnetic field. For example, if electrons (blue) are kicked out by the star, positrons (in red) hit the star’s magnetic poles like in an ‘own goal’. Panel (c) illustrates the size of Geminga’s magnetic field (blue) compared to that of the star itself at the centre (purple). The magnetic field is tilted with respect to Geminga’s rotation axis (red). Panel (d) shows the magnetic poles of Geminga, where charged particles hit the surface of the star, creating a two-million degrees hot spot, a region much hotter than the surroundings. As the star spins on its rotation axis, the hot spot comes into view and then disappears, causing the periodic colour change seen by XMM-Newton. An animated version of the entire sequence can be found at: Click here for animated GIF [low resolution, animated GIF, 5536 KB] Click here for AVI [high resolution, AVI with DIVX compression, 19128 KB] hi-res Size hi-res: 371 kb Credits: ESA, P. Caraveo (IASF, Milan) Geminga's hot spot, panel (a) Panel (a) shows an image taken with the EPIC instrument on board the XMM-Newton observatory. The bright tails, made of

Rocky Mountain Spotted Fever

Science.gov (United States)

... with facebook share with twitter share with linkedin Rocky Mountain Spotted Fever Credit: CDC A male cayenne tick, Amblyomma cajennense, ... and New Mexico. Why Is the Study of Rocky Mountain Spotted Fever a Priority for NIAID? Tickborne diseases are becoming ...

SPOT: the door to digital innovation

International Nuclear Information System (INIS)

Anon.

2017-01-01

AREVA NP has opened in Lyon a small department, named SPOT, whose aim is to develop innovative digital technologies such as CAVE and MODOP'3D. CAVE, that is the acronym of Cave Automatic Virtual Environment, is a large screen that splits the room in half and shows the virtual copy of the Astrid reactor. Viewers can wander inside the reactor and its building through the screen and sees all the details of the design. CAVE represents a complete virtual 3-dimension prototype of the Astrid reactor whose aim is to help assessing any change in the design. MODOP'3D is a virtual maintenance tool that allows the operator to simulate reactor maintenance operations in a 3-dimensional way in order to assess their feasibility and optimize them. Exoskeletons whose purpose is to help operators to handle heavy loads are also tested at SPOT. (A.C.)

A Novel Detection Platform for Shrimp White Spot Syndrome Virus Using an ICP11-Dependent Immunomagnetic Reduction (IMR) Assay.

Science.gov (United States)

Liu, Bing-Hsien; Lin, Yu-Chen; Ho, Chia-Shin; Yang, Che-Chuan; Chang, Yun-Tsui; Chang, Jui-Feng; Li, Chun-Yuan; Cheng, Cheng-Shun; Huang, Jiun-Yan; Lee, Yen-Fu; Hsu, Ming-Hung; Lin, Feng-Chun; Wang, Hao-Ching; Lo, Chu-Fang; Yang, Shieh-Yueh; Wang, Han-Ching

2015-01-01

Shrimp white spot disease (WSD), which is caused by white spot syndrome virus (WSSV), is one of the world's most serious shrimp diseases. Our objective in this study was to use an immunomagnetic reduction (IMR) assay to develop a highly sensitive, automatic WSSV detection platform targeted against ICP11 (the most highly expressed WSSV protein). After characterizing the magnetic reagents (Fe3O4 magnetic nanoparticles coated with anti ICP11), the detection limit for ICP11 protein using IMR was approximately 2 x 10(-3) ng/ml, and the linear dynamic range of the assay was 0.1~1 x 10(6) ng/ml. In assays of ICP11 protein in pleopod protein lysates from healthy and WSSV-infected shrimp, IMR signals were successfully detected from shrimp with low WSSV genome copy numbers. We concluded that this IMR assay targeting ICP11 has potential for detecting the WSSV.

Extended morphological processing: a practical method for automatic spot detection of biological markers from microscopic images.

Science.gov (United States)

Kimori, Yoshitaka; Baba, Norio; Morone, Nobuhiro

2010-07-08

A reliable extraction technique for resolving multiple spots in light or electron microscopic images is essential in investigations of the spatial distribution and dynamics of specific proteins inside cells and tissues. Currently, automatic spot extraction and characterization in complex microscopic images poses many challenges to conventional image processing methods. A new method to extract closely located, small target spots from biological images is proposed. This method starts with a simple but practical operation based on the extended morphological top-hat transformation to subtract an uneven background. The core of our novel approach is the following: first, the original image is rotated in an arbitrary direction and each rotated image is opened with a single straight line-segment structuring element. Second, the opened images are unified and then subtracted from the original image. To evaluate these procedures, model images of simulated spots with closely located targets were created and the efficacy of our method was compared to that of conventional morphological filtering methods. The results showed the better performance of our method. The spots of real microscope images can be quantified to confirm that the method is applicable in a given practice. Our method achieved effective spot extraction under various image conditions, including aggregated target spots, poor signal-to-noise ratio, and large variations in the background intensity. Furthermore, it has no restrictions with respect to the shape of the extracted spots. The features of our method allow its broad application in biological and biomedical image information analysis.

Laser Pyrometer For Spot Temperature Measurements

Science.gov (United States)

Elleman, D. D.; Allen, J. L.; Lee, M. C.

1988-01-01

Laser pyrometer makes temperature map by scanning measuring spot across target. Scanning laser pyrometer passively measures radiation emitted by scanned spot on target and calibrated by similar passive measurement on blackbody of known temperature. Laser beam turned on for active measurements of reflectances of target spot and reflectance standard. From measurements, temperature of target spot inferred. Pyrometer useful for non-contact measurement of temperature distributions in processing of materials.

Maternal serum proteome changes between the first and third trimester of pregnancy in rural southern Nepal.

Science.gov (United States)

Scholl, P F; Cole, R N; Ruczinski, I; Gucek, M; Diez, R; Rennie, A; Nathasingh, C; Schulze, K; Christian, P; Yager, J D; Groopman, J D; West, K P

2012-05-01

Characterization of normal changes in the serum proteome during pregnancy may enhance understanding of maternal physiology and lead to the development of new gestational biomarkers. In 23 Nepalese pregnant women who delivered at term, two-dimensional difference in-gel electrophoresis (DIGE) was used to assess changes in relative protein abundance between paired serum samples collected in the first and third trimesters. One-hundred and forty-five of over 700 protein spots in DIGE gels (pI 4.2-6.8) exhibited nominally significant (p < 0.05) differences in abundance across trimesters. Additional filtering using a Bonferroni correction reduced the number of significant (p < 0.00019) spots to 61. Mass spectrometric analysis detected 38 proteins associated with gestational age, cytoskeletal remodeling, blood pressure regulation, lipid and nutrient transport, and inflammation. One new protein, pregnancy-specific β-glycoprotein 4 was detected. A follow-up isotope tagging for relative and absolute quantitation (iTRAQ) experiment of six mothers from the DIGE study revealed 111 proteins, of which 11 exhibited significant (p < 0.05) differences between trimesters. Four of these proteins: gelsolin, complement C1r subcomponent, α-1-acid glycoprotein, and α-1B-glycoprotein also changed in the DIGE analysis. Although not previously associated with normal pregnancy, gelsolin decreased in abundance by the third trimester (p < 0.01) in DIGE, iTRAQ and Western analyses. Changes in abundance of proteins in serum that are associated with syncytiotrophoblasts (gelsolin, pregnancy-specific β-1 glycoprotein 1 and β-2-glycoprotein I) probably reflect dynamics of a placental proteome shed into maternal circulation during pregnancy. Measurement of changes in the maternal serum proteome, when linked with birth outcomes, may yield biomarkers for tracking reproductive health in resource poor settings in future studies. Published by Elsevier Ltd.

Tuning and Switching Enantioselectivity of Asymmetric Carboligation in an Enzyme through Mutational Analysis of a Single Hot Spot.

Science.gov (United States)

Wechsler, Cindy; Meyer, Danilo; Loschonsky, Sabrina; Funk, Lisa-Marie; Neumann, Piotr; Ficner, Ralf; Brodhun, Florian; Müller, Michael; Tittmann, Kai

2015-12-01

Enantioselective bond making and breaking is a hallmark of enzyme action, yet switching the enantioselectivity of the reaction is a difficult undertaking, and typically requires extensive screening of mutant libraries and multiple mutations. Here, we demonstrate that mutational diversification of a single catalytic hot spot in the enzyme pyruvate decarboxylase gives access to both enantiomers of acyloins acetoin and phenylacetylcarbinol, important pharmaceutical precursors, in the case of acetoin even starting from the unselective wild-type protein. Protein crystallography was used to rationalize these findings and to propose a mechanistic model of how enantioselectivity is controlled. In a broader context, our studies highlight the efficiency of mechanism-inspired and structure-guided rational protein design for enhancing and switching enantioselectivity of enzymatic reactions, by systematically exploring the biocatalytic potential of a single hot spot. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

KFC2: a knowledge-based hot spot prediction method based on interface solvation, atomic density, and plasticity features.

Science.gov (United States)

Zhu, Xiaolei; Mitchell, Julie C

2011-09-01

Hot spots constitute a small fraction of protein-protein interface residues, yet they account for a large fraction of the binding affinity. Based on our previous method (KFC), we present two new methods (KFC2a and KFC2b) that outperform other methods at hot spot prediction. A number of improvements were made in developing these new methods. First, we created a training data set that contained a similar number of hot spot and non-hot spot residues. In addition, we generated 47 different features, and different numbers of features were used to train the models to avoid over-fitting. Finally, two feature combinations were selected: One (used in KFC2a) is composed of eight features that are mainly related to solvent accessible surface area and local plasticity; the other (KFC2b) is composed of seven features, only two of which are identical to those used in KFC2a. The two models were built using support vector machines (SVM). The two KFC2 models were then tested on a mixed independent test set, and compared with other methods such as Robetta, FOLDEF, HotPoint, MINERVA, and KFC. KFC2a showed the highest predictive accuracy for hot spot residues (True Positive Rate: TPR = 0.85); however, the false positive rate was somewhat higher than for other models. KFC2b showed the best predictive accuracy for hot spot residues (True Positive Rate: TPR = 0.62) among all methods other than KFC2a, and the False Positive Rate (FPR = 0.15) was comparable with other highly predictive methods. Copyright © 2011 Wiley-Liss, Inc.

Forecasting European thermal coal spot prices

Directory of Open Access Journals (Sweden)

Alicja Krzemień

2015-01-01

Finally, in order to analyse the time series model performance a Generalized Regression Neural Network (GRNN was used and its performance compared against the whole AR(2 process. Empirical results obtained confirmed that there is no statistically significant difference between both methods. The GRNN analysis also allowed pointing out the main drivers that move the European Thermal Coal Spot prices: crude oil, USD/CNY change and supply side drivers.

THE INFORMATION CONTENT IN ANALYTIC SPOT MODELS OF BROADBAND PRECISION LIGHT CURVES

Energy Technology Data Exchange (ETDEWEB)

Walkowicz, Lucianne M. [Department of Astrophysical Sciences, Princeton University, Peyton Hall, 4 Ivy Lane, Princeton, NJ 08534 (United States); Basri, Gibor [Astronomy Department, University of California at Berkeley, Hearst Field Annex, Berkeley, CA 94720 (United States); Valenti, Jeff A. [Space Telescope Science Institute, 3700 San Martin Dr., Baltimore, MD 21218 (United States)

2013-04-01

We present the results of numerical experiments to assess degeneracies in light curve models of starspots. Using synthetic light curves generated with the Cheetah starspot modeling code, we explore the extent to which photometric light curves constrain spot model parameters, including spot latitudes and stellar inclination. We also investigate the effects of spot parameters and differential rotation on one's ability to correctly recover rotation periods and differential rotation in the Kepler light curves. We confirm that in the absence of additional constraints on the stellar inclination, such as spectroscopic measurements of vsin i or occultations of starspots by planetary transits, the spot latitude and stellar inclination are difficult to determine uniquely from the photometry alone. We find that for models with no differential rotation, spots that appear on opposite hemispheres of the star may cause one to interpret the rotation period to be half of the true period. When differential rotation is included, the changing longitude separation between spots breaks the symmetry of the hemispheres and the correct rotation period is more likely to be found. The dominant period found via periodogram analysis is typically that of the largest spot. Even when multiple spots with periods representative of the star's differential rotation exist, if one spot dominates the light curve the signal of differential rotation may not be detectable from the periodogram alone. Starspot modeling is applicable to stars with a wider range of rotation rates than other surface imaging techniques (such as Doppler imaging), allows subtle signatures of differential rotation to be measured, and may provide valuable information on the distribution of stellar spots. However, given the inherent degeneracies and uncertainty present in starspot models, caution should be exercised in their interpretation.

THE INFORMATION CONTENT IN ANALYTIC SPOT MODELS OF BROADBAND PRECISION LIGHT CURVES

International Nuclear Information System (INIS)

Walkowicz, Lucianne M.; Basri, Gibor; Valenti, Jeff A.

2013-01-01

We present the results of numerical experiments to assess degeneracies in light curve models of starspots. Using synthetic light curves generated with the Cheetah starspot modeling code, we explore the extent to which photometric light curves constrain spot model parameters, including spot latitudes and stellar inclination. We also investigate the effects of spot parameters and differential rotation on one's ability to correctly recover rotation periods and differential rotation in the Kepler light curves. We confirm that in the absence of additional constraints on the stellar inclination, such as spectroscopic measurements of vsin i or occultations of starspots by planetary transits, the spot latitude and stellar inclination are difficult to determine uniquely from the photometry alone. We find that for models with no differential rotation, spots that appear on opposite hemispheres of the star may cause one to interpret the rotation period to be half of the true period. When differential rotation is included, the changing longitude separation between spots breaks the symmetry of the hemispheres and the correct rotation period is more likely to be found. The dominant period found via periodogram analysis is typically that of the largest spot. Even when multiple spots with periods representative of the star's differential rotation exist, if one spot dominates the light curve the signal of differential rotation may not be detectable from the periodogram alone. Starspot modeling is applicable to stars with a wider range of rotation rates than other surface imaging techniques (such as Doppler imaging), allows subtle signatures of differential rotation to be measured, and may provide valuable information on the distribution of stellar spots. However, given the inherent degeneracies and uncertainty present in starspot models, caution should be exercised in their interpretation.

Time-varying convergence in European electricity spot markets and their association with carbon and fuel prices

International Nuclear Information System (INIS)

Menezes, Lilian M. de; Houllier, Melanie A.; Tamvakis, Michael

2016-01-01

Long-run dynamics of electricity prices are expected to reflect fuel price developments, since fuels generally account for a large share in the cost of generation. As an integrated European market for electricity develops, wholesale electricity prices should be converging as a result of market coupling and increased interconnectivity. Electricity mixes are also changing, spurred by a drive to significantly increase the share of renewables. Consequently, the electricity wholesale price dynamics are evolving, and the fuel–electricity price nexus that has been described in the literature is likely to reflect this evolution. This study investigates associations between spot prices from the British, French and Nordpool markets with those in connected electricity markets and fuel input prices, from December 2005 to October 2013. In order to assess the time-varying dynamics of electricity spot price series, localized autocorrelation functions are used. Electricity spot prices in the three markets are found to have stationary and non-stationary periods. When a trend in spot prices is observed, it is likely to reflect the trend in fuel prices. Cointegration analysis is then used to assess co-movement between electricity spot prices and fuel inputs to generation. The results show that British electricity spot prices are associated with fuel prices and not with price developments in connected markets, while the opposite is observed in the French and Nordpool day-ahead markets. - Highlights: • Electricity market integration policies may have altered EU spot electricity prices. • LACF is used to assess the changing nature of electricity spot prices. • EU electricity spot prices show both stationary and non-stationary periods. • Carbon and fuel prices have greater impact on British spot prices. • In continental Europe, electricity prices have decoupled from fuel prices.

Resistance Spot Welding of dissimilar Steels

Directory of Open Access Journals (Sweden)

Ladislav Kolařík

2012-01-01

Full Text Available This paper presents an analysis of the properties of resistance spot welds between low carbon steel and austenitic CrNi stainless steel. The thickness of the welded dissimilar materials was 2 mm. A DeltaSpot welding gun with a process tape was used for welding the dissimilar steels. Resistance spot welds were produced with various welding parameters (welding currents ranging from 7 to 8 kA. Light microscopy, microhardness measurements across the welded joints, and EDX analysis were used to evaluate the quality of the resistance spot welds. The results confirm the applicability of DeltaSpot welding for this combination of materials.

Oil futures and spot markets

International Nuclear Information System (INIS)

Samii, M.V.

1992-01-01

In the last decade, the oil futures market has risen to prominence and has become a major factor in influencing oil market psychology and the crude oil market. On a normal day, over 92 thousand contracts, the equivalent of 92 million barrels per day, change hands on the New York Mercantile Exchange, NYMEX. This market has provided a vehicle for hedging against risk. At the same time, it has also created opportunities for speculation. Those who previously were unable to participate in oil market transactions can now become involved through the futures market. The large number of participants in the future market and the availability of information has made this market more efficient and transparent, relative to the crude oil market. While there has been considerable in-depth analysis of other future markets, relatively little theoretical attention has focused on that of oil. This paper looks at the following issues. First, what is the relationship between futures and spot oil prices? And secondly, are futures prices a good predictor of spot crude prices in the future? (author)

Eczematous Dermatitis Occurring on a Café-au-Lait Spot Long after Laser Radiation.

Science.gov (United States)

Mihara, Motoyuki

2013-05-01

A 40-year-old woman presented with an itchy erythematosquamous change of a café-au-lait spot in her face. The onset of this change occurred just after her relocation. The café-au-lait spot had been irradiated by laser approximately 20 years ago. Clinically, there was a coin-sized erythema with a slight scale on the pigmented lesion in the left lateral orbital region. Histopathologically, the lesion demonstrated both spongiotic dermatitis and interface dermatitis together with lymphohistiocytic cell infiltration, in addition to moderate acanthosis and elongation of rete ridges with slight basal hyperpigmentation. From these clinical and histopathological findings, the lesion was diagnosed as eczematous dermatitis occurring on the café-au-lait spot after laser radiation. Another interesting histopathological finding was that some parts of a lobule of the sebaceous gland were occupied exclusively by degenerative atrophic sebocytes. From the viewpoint of pathogenesis, the eczematous dermatitis of this patient could have been an accompanying feature of a neurogenic inflammation occurring on the café-au-lait spot after laser radiation, and the atrophic change of a part of the sebaceous lobule might have been induced by a morphogenetic alteration of certain germinative cells of the sebaceous lobule due to laser radiation.

On the relationships between electron spot size, focal spot size, and virtual source position in Monte Carlo simulations

International Nuclear Information System (INIS)

Sterpin, E.; Chen, Y.; Lu, W.; Mackie, T. R.; Olivera, G. H.; Vynckier, S.

2011-01-01

Purpose: Every year, new radiotherapy techniques including stereotactic radiosurgery using linear accelerators give rise to new applications of Monte Carlo (MC) modeling. Accurate modeling requires knowing the size of the electron spot, one of the few parameters to tune in MC models. The resolution of integrated megavoltage imaging systems, such as the tomotherapy system, strongly depends on the photon spot size which is closely related to the electron spot. The aim of this article is to clarify the relationship between the electron spot size and the photon spot size (i.e., the focal spot size) for typical incident electron beam energies and target thicknesses. Methods: Three electron energies (3, 5.5, and 18 MeV), four electron spot sizes (FWHM=0, 0.5, 1, and 1.5 mm), and two tungsten target thicknesses (0.15 and 1 cm) were considered. The formation of the photon beam within the target was analyzed through electron energy deposition with depth, as well as photon production at several phase-space planes placed perpendicular to the beam axis, where only photons recorded for the first time were accounted for. Photon production was considered for ''newborn'' photons intersecting a 45x45 cm 2 plane at the isocenter (85 cm from source). Finally, virtual source position and ''effective'' focal spot size were computed by backprojecting all the photons from the bottom of the target intersecting a 45x45 cm 2 plane. The virtual source position and focal spot size were estimated at the plane position where the latter is minimal. Results: In the relevant case of considering only photons intersecting the 45x45 cm 2 plane, the results unambiguously showed that the effective photon spot is created within the first 0.25 mm of the target and that electron and focal spots may be assumed to be equal within 3-4%. Conclusions: In a good approximation photon spot size equals electron spot size for high energy X-ray treatments delivered by linear accelerators.

Proteomic analysis of pregnancy-related proteins from pig uterus endometrium during pregnancy

Directory of Open Access Journals (Sweden)

Kang Sunghyun

2011-07-01

Full Text Available Abstract Many important molecular events associated with implantation and development occur within the female reproductive tract, especially within the uterus endometrium, during pregnancy periods. The endometrium includes the mucosal lining of the uterus, which provides a suitable site for implantation and development of a fertilized egg and fetus. To date, the molecular cascades in the uterus endometrium during pregnancy periods in pigs have not been elucidated fully. In this study, we compared the functional regulated proteins in the endometrium during pregnancy periods with those in non-pregnant conditions and investigated changes in expression patterns during pregnancy (days 40, 70, and 93 using two-dimensional gel electrophoresis (2-DE and western blotting. The functional regulated proteins were identified and discovered from differentially expressed proteins in the uterus endometrium during pregnancy. We discovered 820 protein spots in a proteomic analysis of uterus endometrium tissues with 2-DE gels. We identified 63 of the 98 proteins regulated differentially among non-pregnant and pregnant tissues (matched and unmatched spots. Interestingly, 10 of these 63 proteins are development-, cytoskeleton- and chaperon-related proteins such as transferrin, protein DJ-1, transgelin, galectin-1, septin 2, stathmin 1, cofilin 1, fascin 1, heat shock protein (HSP 90β and HSP 27. The specific expression patterns of these proteins in the endometrium during pregnancy were confirmed by western blotting. Our results suggest that the expressions of these genes involved in endometrium function and endometrium development from early to late gestation are associated with the regulation of endometrium development for maintaining pregnancy.

Multivariate data analysis of two-dimensional gel electrophoresis protein patterns from few samples

DEFF Research Database (Denmark)

Jensen, Kristina Nedenskov; Jessen, Flemming; Jørgensen, Bo

2008-01-01

One application of 2D gel electrophoresis is to reveal differences in protein pattern between two or more groups of individuals, attributable to their group membership. Multivariate data analytical methods are useful in pinpointing the spots relevant for discrimination by focusing not only...... on single spot differences, but on the covariance structure between proteins. However, their outcome is dependent on data scaling, and they may fail in producing valid multivariate models due to the much higher number of "irrelevant" spots present in the gels. The case where only few gels are available...... and where the aim is to find as many as possible of the group-dependent proteins seems particularly difficult to handle. The present paper investigates such a case regarding the effect of scaling and of prefiltering by univariate nonparametric statistics on the selection of spots. Besides, a modified...

Evidence of high-field radio-frequency hot spots due to trapped vortices in niobium cavities

Directory of Open Access Journals (Sweden)

G. Ciovati

2008-12-01

Full Text Available Superconducting radio-frequency (rf cavities made of high-purity niobium exhibit strong anomalous rf losses starting at peak surface magnetic fields of about 90–100 mT in the gigahertz range. This phenomenon is referred to as “Q drop.” Temperature maps of the cavity surface have revealed the presence of “hot spots” in the high magnetic field region of the cavities. Several models have been proposed over the years to explain this phenomenon but there is still no experimental evidence on the mechanisms behind such hot spots. In this work we show that at least some of the hot spots are due to trapped vortices responsible for the anomalous losses. Here we report experiments in which a local thermal gradient was applied to the hot spot regions of a cavity in order to displace the vortices. Temperature maps measured before and after applying the thermal gradient unambiguously show that the hot spots do move and change their intensities, allowing us to determine changes in the hot spot positions and strengths and their effect on the cavity performance. Results on a large-grain niobium cavity clearly show a different distribution and in some cases a weakening of the intensity of the “hot spots,” suggesting new ways of improving the cavity performance without additional material treatments.

Effect of Crossflow on Hot Spot Fuel Temperature in Prismatic VHTR

International Nuclear Information System (INIS)

Lee, Sung Nam; Tak, Nam-il; Kim, Min Hwan; Noh, Jae Man; Park, Goon-Cherl

2014-01-01

Various studies have been conducted to predict the thermal-hydraulics of a prismatic gas-cooled reactor. However, most previous studies have concentrated on the nominal-designed core. The fuel assembly of a high temperature gas-cooled reactor consists of a fuel compact and graphite block used as a moderator. This graphite faces a dimensional change due to irradiation or heating during normal operation. This size change might affect the coolant flow distribution in the active core. Therefore, the hot spot fuel temperature position or value could vary. There are two types of flows by the size change of graphite. One is the bypass flow and the other is the crossflow. The crossflow occurs at the crossflow gap between the vertical stacks of fuel blocks. In this study, the effect of the crossflow on the hot spot fuel temperature has been intensively investigated. (author)

Solar 'hot spots' are still hot

Science.gov (United States)

Bai, Taeil

1990-01-01

Longitude distributions of solar flares are not random but show evidence for active zones (or hot spots) where flares are concentrated. According to a previous study, two hot spots in the northern hemisphere, which rotate with a synodic period of about 26.72 days, produced the majority of major flares, during solar cycles 20 and 21. The more prominent of these two hot spots is found to be still active during the rising part of cycle 22, producing the majority of northern hemisphere major flares. The synodic rotation period of this hot spot is 26.727 + or - 0.007 days. There is also evidence for hot spots in the southern hemisphere. Two hot spots separated by 180 deg are found to rotate with a period of 29.407 days, with one of them having persisted in the same locations during cycles 19-22 and the other, during cycles 20-22.

Age-dependent changes in the total protein concentrations in the ...

African Journals Online (AJOL)

related changes in total protein concentrations in ten regions of the pig brain and hypophyses from birth to 36 months of age. Age-related changes in protein concentrations in all the brain regions except the pons and cerebral cortex were not ...

In Situ Hot-Spot Assembly as a General Strategy for Probing Single Biomolecules.

Science.gov (United States)

Liu, Huiqiao; Li, Qiang; Li, Mingmin; Ma, Sisi; Liu, Dingbin

2017-05-02

Single-molecule detection using surface-enhanced Raman spectroscopy (SERS) has attracted increasing attention in chemical and biomedical analysis. However, it remains a major challenge to probe single biomolecules by means of SERS hot spots owing to the small volume of hot spots and their random distribution on substrates. We here report an in situ hot-spot assembly method as a general strategy for probing single biomolecules. As a proof-of-concept, this proposed strategy was successfully used for the detection of single microRNA-21 (miRNA-21, a potential cancer biomarker) at the single-cell level, showing great capability in differentiating the expression of miRNA-21 in single cancer cells from normal cells. This approach was further extended to single-protein detection. The versatility of the strategy opens an exciting avenue for single-molecule detection of biomarkers of interest and thus holds great promise in a variety of biological and biomedical applications.

Differential Protein Expression in Congenital and Acquired Cholesteatomas.

Directory of Open Access Journals (Sweden)

Seung-Ho Shin

Full Text Available Congenital cholesteatomas are epithelial lesions that present as an epithelial pearl behind an intact eardrum. Congenital and acquired cholesteatomas progress quite differently from each other and progress patterns can provide clues about the unique origin and pathogenesis of the abnormality. However, the exact pathogenic mechanisms by which cholesteatomas develop remain unknown. In this study, key proteins that directly affect cholesteatoma pathogenesis are investigated with proteomics and immunohistochemistry. Congenital cholesteatoma matrices and retroauricular skin were harvested during surgery in 4 patients diagnosed with a congenital cholesteatoma. Tissue was also harvested from the retraction pocket in an additional 2 patients during middle ear surgery. We performed 2-dimensional (2D electrophoresis to detect and analyze spots that are expressed only in congenital cholesteatoma and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/MS to separate proteins by molecular weight. Protein expression was confirmed by immunohistochemical staining. The image analysis of 2D electrophoresis showed that 4 congenital cholesteatoma samples had very similar protein expression patterns and that 127 spots were exclusively expressed in congenital cholesteatomas. Of these 127 spots, 10 major spots revealed the presence of titin, forkhead transcription activator homolog (FKH 5-3, plectin 1, keratin 10, and leucine zipper protein 5 by MALDI-TOF/MS analysis. Immunohistochemical staining showed that FKH 5-3 and titin were expressed in congenital cholesteatoma matrices, but not in acquired cholesteatomas. Our study shows that protein expression patterns are completely different in congenital cholesteatomas, acquired cholesteatomas, and skin. Moreover, non-epithelial proteins, including FKH 5-3 and titin, were unexpectedly expressed in congenital cholesteatoma tissue. Our data indicates that congenital cholesteatoma origins

Arc cathode spots

International Nuclear Information System (INIS)

Schrade, H.O.

1989-01-01

Arc spots are usually highly unstable and jump statistically over the cathode surface. In a magnetic field parallel to the surface, preferably they move in the retrograde direction; i.e., opposite to the Lorentzian rule. If the field is inclined with respect to the surface, the spots drift away at a certain angle with respect to the proper retrograde direction (Robson drift motion). These well-known phenomena are explained by one stability theory

7 CFR 28.413 - Middling Light Spotted Color.

Science.gov (United States)

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Light Spotted Color. 28.413 Section 28.413... Spotted Color. Middling Light Spotted Color is color which in spot or color, or both, is between Middling Color and Middling Spotted Color. ...

Detecting significant changes in protein abundance

Directory of Open Access Journals (Sweden)

Kai Kammers

2015-06-01

Full Text Available We review and demonstrate how an empirical Bayes method, shrinking a protein's sample variance towards a pooled estimate, leads to far more powerful and stable inference to detect significant changes in protein abundance compared to ordinary t-tests. Using examples from isobaric mass labelled proteomic experiments we show how to analyze data from multiple experiments simultaneously, and discuss the effects of missing data on the inference. We also present easy to use open source software for normalization of mass spectrometry data and inference based on moderated test statistics.

Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis

Science.gov (United States)

Lee, Jae Eun; Lee, Jae Young; Kim, Hong Rye; Shin, Hyun Young; Lin, Tao; Jin, Dong Il

2015-01-01

Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum. PMID:25925056

Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis

Directory of Open Access Journals (Sweden)

Jae Eun Lee

2015-06-01

Full Text Available Two dimensional-fluorescence difference gel electrophoresis (2D DIGE is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

Changes in Proteome Profile of Peripheral Blood Mononuclear Cells in Chronic Chagas Disease.

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Nisha Jain Garg

2016-02-01

Full Text Available Trypanosoma cruzi (Tc infection causes chagasic cardiomyopathy; however, why 30-40% of the patients develop clinical disease is not known. To discover the pathomechanisms in disease progression, we obtained the proteome signature of peripheral blood mononuclear cells (PBMCs of normal healthy controls (N/H, n = 30 and subjects that were seropositive for Tc-specific antibodies, but were clinically asymptomatic (C/A, n = 25 or clinically symptomatic (C/S, n = 28 with cardiac involvement and left ventricular dysfunction. Protein samples were labeled with BODIPY FL-maleimide (dynamic range: > 4 orders of magnitude, detection limit: 5 f-mol and resolved by two-dimensional gel electrophoresis (2D-GE. After normalizing the gel images, protein spots that exhibited differential abundance in any of the two groups were analyzed by mass spectrometry, and searched against UniProt human database for protein identification. We found 213 and 199 protein spots (fold change: |≥ 1.5|, p93% prediction success in classifying infected individuals with no disease and those with cardiac involvement and LV dysfunction. In conclusion, we have identified molecular pathways and a panel of proteins that could aid in detecting seropositive individuals at risk of developing cardiomyopathy.

Bright spots among the world’s coral reefs

Science.gov (United States)

Cinner, Joshua E.; Huchery, Cindy; MacNeil, M. Aaron; Graham, Nicholas A. J.; McClanahan, Tim R.; Maina, Joseph; Maire, Eva; Kittinger, John N.; Hicks, Christina C.; Mora, Camilo; Allison, Edward H.; D'Agata, Stephanie; Hoey, Andrew; Feary, David A.; Crowder, Larry; Williams, Ivor D.; Kulbicki, Michel; Vigliola, Laurent; Wantiez, Laurent; Edgar, Graham; Stuart-Smith, Rick D.; Sandin, Stuart A.; Green, Alison L.; Hardt, Marah J.; Beger, Maria; Friedlander, Alan; Campbell, Stuart J.; Holmes, Katherine E.; Wilson, Shaun K.; Brokovich, Eran; Brooks, Andrew J.; Cruz-Motta, Juan J.; Booth, David J.; Chabanet, Pascale; Gough, Charlie; Tupper, Mark; Ferse, Sebastian C. A.; Sumaila, U. Rashid; Mouillot, David

2016-07-01

Ongoing declines in the structure and function of the world’s coral reefs require novel approaches to sustain these ecosystems and the millions of people who depend on them. A presently unexplored approach that draws on theory and practice in human health and rural development is to systematically identify and learn from the ‘outliers’—places where ecosystems are substantially better (‘bright spots’) or worse (‘dark spots’) than expected, given the environmental conditions and socioeconomic drivers they are exposed to. Here we compile data from more than 2,500 reefs worldwide and develop a Bayesian hierarchical model to generate expectations of how standing stocks of reef fish biomass are related to 18 socioeconomic drivers and environmental conditions. We identify 15 bright spots and 35 dark spots among our global survey of coral reefs, defined as sites that have biomass levels more than two standard deviations from expectations. Importantly, bright spots are not simply comprised of remote areas with low fishing pressure; they include localities where human populations and use of ecosystem resources is high, potentially providing insights into how communities have successfully confronted strong drivers of change. Conversely, dark spots are not necessarily the sites with the lowest absolute biomass and even include some remote, uninhabited locations often considered near pristine. We surveyed local experts about social, institutional, and environmental conditions at these sites to reveal that bright spots are characterized by strong sociocultural institutions such as customary taboos and marine tenure, high levels of local engagement in management, high dependence on marine resources, and beneficial environmental conditions such as deep-water refuges. Alternatively, dark spots are characterized by intensive capture and storage technology and a recent history of environmental shocks. Our results suggest that investments in strengthening fisheries

Xanthomonas euvesicatoria Causes Bacterial Spot Disease on Pepper Plant in Korea

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Min-Seong Kyeon

2016-10-01

Full Text Available In 2004, bacterial spot-causing xanthomonads (BSX were reclassified into 4 species—Xanthomonas euvesicatoria, X. vesicatoria, X. perforans, and X. gardneri. Bacterial spot disease on pepper plant in Korea is known to be caused by both X. axonopodis pv. vesicatoria and X. vesicatoria. Here, we reidentified the pathogen causing bacterial spots on pepper plant based on the new classification. Accordingly, 72 pathogenic isolates were obtained from the lesions on pepper plants at 42 different locations. All isolates were negative for pectolytic activity. Five isolates were positive for amylolytic activity. All of the Korean pepper isolates had a 32 kDa-protein unique to X. euvesicatoria and had the same band pattern of the rpoB gene as that of X. euvesicatoria and X. perforans as indicated by PCR-restriction fragment length polymorphism analysis. A phylogenetic tree of 16S rDNA sequences showed that all of the Korean pepper plant isolates fit into the same group as did all the reference strains of X. euvesicatoria and X. perforans. A phylogenetic tree of the nucleotide sequences of 3 housekeeping genes—gapA, gyrB, and lepA showed that all of the Korean pepper plant isolates fit into the same group as did all of the references strains of X. euvesicatoria. Based on the phenotypic and genotypic characteristics, we identified the pathogen as X. euvesicatoria. Neither X. vesicatoria, the known pathogen of pepper bacterial spot, nor X. perforans, the known pathogen of tomato plant, was isolated. Thus, we suggest that the pathogen causing bacterial spot disease of pepper plants in Korea is X. euvesicatoria.

Solar hot spots are still hot

International Nuclear Information System (INIS)

Bai, T.

1990-01-01

Longitude distributions of solar flares are not random but show evidence for active zones (or hot spots) where flares are concentrated. According to a previous study, two hot spots in the northern hemisphere, which rotate with a synodic period of about 26.72 days, produced the majority of major flares, during solar cycles 20 and 21. The more prominent of these two hot spots is found to be still active during the rising part of cycle 22, producing the majority of northern hemisphere major flares. The synodic rotation period of this hot spot is 26.727 + or - 0.007 days. There is also evidence for hot spots in the southern hemisphere. Two hot spots separated by 180 deg are found to rotate with a period of 29.407 days, with one of them having persisted in the same locations during cycles 19-22 and the other, during cycles 20-22. 14 refs

Experimental and simulated strength of spot welds

DEFF Research Database (Denmark)

Nielsen, Chris Valentin; Bennedbæk, Rune A.K.; Larsen, Morten B.

2014-01-01

Weld strength testing of single spots in DP600 steel is presented for the three typical testing procedures, i.e. tensile-shear, cross-tension and peel testing. Spot welds are performed at two sets of welding parameters and strength testing under these conditions is presented by load......-elongation curves revealing the maximum load and the elongation at break. Welding and strength testing is simulated by SORPAS® 3D, which allows the two processes to be prepared in a combined simulation, such that the simulated welding properties are naturally applied to the simulation of strength testing. Besides...... the size and shape of the weld nugget, these properties include the new strength of the material in the weld and the heat affected zone based on the predicted hardness resulting from microstructural phase changes simulated during cooling of the weld before strength testing. Comparisons between overall...

7 CFR 28.423 - Middling Spotted Color.

Science.gov (United States)

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody of...

The role of relA and spoT in Yersinia pestis KIM5 pathogenicity.

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Wei Sun

2009-08-01

Full Text Available The ppGpp molecule is part of a highly conserved regulatory system for mediating the growth response to various environmental conditions. This mechanism may represent a common strategy whereby pathogens such as Yersinia pestis, the causative agent of plague, regulate the virulence gene programs required for invasion, survival and persistence within host cells to match the capacity for growth. The products of the relA and spoT genes carry out ppGpp synthesis. To investigate the role of ppGpp on growth, protein synthesis, gene expression and virulence, we constructed a Delta relA Delta spoT Y. pestis mutant. The mutant was no longer able to synthesize ppGpp in response to amino acid or carbon starvation, as expected. We also found that it exhibited several novel phenotypes, including a reduced growth rate and autoaggregation at 26 degrees C. In addition, there was a reduction in the level of secretion of key virulence proteins and the mutant was > 1,000-fold less virulent than its wild-type parent strain. Mice vaccinated subcutaneously (s.c. with 2.5x10(4 CFU of the Delta relA Delta spoT mutant developed high anti-Y. pestis serum IgG titers, were completely protected against s.c. challenge with 1.5x10(5 CFU of virulent Y. pestis and partially protected (60% survival against pulmonary challenge with 2.0x10(4 CFU of virulent Y. pestis. Our results indicate that ppGpp represents an important virulence determinant in Y. pestis and the Delta relA Delta spoT mutant strain is a promising vaccine candidate to provide protection against plague.

Mercury Exposure: Protein Biomarkers of Mercury Exposure in Jaraqui Fish from the Amazon Region.

Science.gov (United States)

Vieira, José Cavalcante Souza; Braga, Camila Pereira; de Oliveira, Grasieli; Padilha, Cilene do Carmo Federici; de Moraes, Paula Martin; Zara, Luiz Fabricio; Leite, Aline de Lima; Buzalaf, Marília Afonso Rabelo; Padilha, Pedro de Magalhães

2018-05-01

This study presents data on the extraction and characterization of proteins associated with mercury in the muscle and liver tissues of jaraqui (Semaprochilodus spp.) from the Madeira River in the Brazilian Amazon. Protein fractionation was carried out by two-dimensional electrophoresis (2D-PAGE). Mercury determination in tissues, pellets, and protein spots was performed by graphite furnace atomic absorption spectrometry (GFAAS). Proteins in the spots that showed mercury were characterized by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The highest mercury concentrations were found in liver tissues and pellets (426 ± 6 and 277 ± 4 μg kg -1 ), followed by muscle tissues and pellets (132 ± 4 and 86 ± 1 μg kg -1 , respectively). Mercury quantification in the protein spots allowed us to propose stoichiometric ratios in the range of 1-4 mercury atoms per molecule of protein in the protein spots. The proteins characterized in the analysis by ESI-MS/MS were keratin, type II cytoskeletal 8, parvalbumin beta, parvalbumin-2, ubiquitin-40S ribosomal S27a, 39S ribosomal protein L36 mitochondrial, hemoglobin subunit beta, and hemoglobin subunit beta-A/B. The results suggest that proteins such as ubiquitin-40S ribosomal protein S27a, which have specific domains, possibly zinc finger, can be used as biomarkers of mercury, whereas mercury and zinc present characteristics of soft acids.

Improved simulation method of automotive spot weld failure with an account of the mechanical properties of spot welds

Science.gov (United States)

Wu, H.; Meng, X. M.; Fang, R.; Huang, Y. F.; Zhan, S.

2017-12-01

In this paper, the microstructure and mechanical properties of spot weld were studied, the hardness of nugget and heat affected zone (HAZ) were also tested by metallographic microscope and microhardness tester. The strength of the spot weld with the different parts' area has been characterized. According to the experiments result, CAE model of spot weld with HAZ structure was established, and simulation results of different lap-shear CAE models were analyzed. The results show that the spot weld model which contained the HAZ has good performance and more suitable for engineering application in spot weld simulation.

Proteomic changes in Debaryomyces hansenii upon exposure to NaCl stress

DEFF Research Database (Denmark)

Gori, Klaus; Hébraud, Michel; Chambon, Christophe

2007-01-01

The proteome of the highly NaCl-tolerant yeast Debaryomyces hansenii was investigated by two-dimensional polyacrylamide gel electrophoresis (2D PAGE), and 47 protein spots were identified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) followed by mass spectrometry (MS...... 7% and 4% of the rate in medium without NaCl. In addition, the number of protein spots detected on 2D gels prepared from cells exposed to 8% and 12% (w/v) NaCl exceeded less than 28% of the number of protein spots detected on 2D gels prepared from cells without added NaCl. Several proteins were...

Tomato Spotted Wilt Virus NSs Protein Supports Infection and Systemic Movement of a Potyvirus and Is a Symptom Determinant.

Science.gov (United States)

Garcia-Ruiz, Hernan; Gabriel Peralta, Sergio M; Harte-Maxwell, Patricia A

2018-03-14

Plant viruses are inducers and targets of antiviral RNA silencing. To condition susceptibility, most plant viruses encode silencing suppressor proteins that interfere with antiviral RNA silencing. The NSs protein is an RNA silencing suppressor in orthotospoviruses, such as the tomato spotted wilt virus (TSWV). The mechanism of RNA silencing suppression by NSs and its role in virus infection and movement are poorly understood. Here, we cloned and tagged TSWV NSs and expressed it from a GFP-tagged turnip mosaic virus (TuMV-GFP) carrying either a wild-type or suppressor-deficient (AS9) helper component proteinase (HC-Pro). When expressed in cis, NSs restored pathogenicity and promoted systemic infection of suppressor-deficient TuMV-AS9-GFP in Nicotiana benthamiana and Arabidopsis thaliana . Inactivating mutations were introduced in NSs RNA-binding domain one. A genetic analysis with active and suppressor-deficient NSs, in combination with wild-type and mutant plants lacking essential components of the RNA silencing machinery, showed that the NSs insert is stable when expressed from a potyvirus. NSs can functionally replace potyviral HC-Pro, condition virus susceptibility, and promote systemic infection and symptom development by suppressing antiviral RNA silencing through a mechanism that partially overlaps that of potyviral HC-Pro. The results presented provide new insight into the mechanism of silencing suppression by NSs and its effect on virus infection.

TV spots' impact.

Science.gov (United States)

El-bakly, S

1994-09-01

The Information, Education and Communication (IEC) Center of the State Information Service was established in 1979 for the purpose of providing information to the people on the population issue. The Ministry of Information has accorded the State Information Service free TV and radio air time for family planning dramas and spots. In the early years information campaigns were organized to make people aware of the population problem by slogans, songs, and cartoons. Around 1984 misconceptions about family planning and contraceptives were attacked through a number of TV and radio spots. A few years later 21 spots on specific contraceptive methods were broadcast which were aired for three years over 3000 times. They were extremely successful. The impact of these TV spots was one of the major reasons why the contraceptive prevalence rate increased from 30% in 1984 to 38% in 1988 and 47% in 1992. Spots were also broadcast about the social implications of large families. The TV soap opera "And The Nile Flows On", with the family planning message interwoven into it, was very well received by the target audience. A program entitled "Wedding of the Month" features couples who know family planning well. The most successful radio program is a 15-20 minute long quiz show for residents of the villages where the Select Villages Project is being implemented. The State Information Service has 60 local information centers in the 26 governorates of Egypt that make plans for the family planning campaign. In 1992 the Minya Initiative, a family planning project was implemented in the Minya Governorate. As a result, the contraceptive prevalence rate rose from 22% to 30% over 18 months. A new project, the Select Village Project, was developed in 1993 that replicates the Minya Initiative on the village level in other governorates. This new project that was implemented in sixteen governorates.

Black-spot poison ivy.

Science.gov (United States)

Schram, Sarah E; Willey, Andrea; Lee, Peter K; Bohjanen, Kimberly A; Warshaw, Erin M

2008-01-01

In black-spot poison ivy dermatitis, a black lacquerlike substance forms on the skin when poison ivy resin is exposed to air. Although the Toxicodendron group of plants is estimated to be the most common cause of allergic contact dermatitis in the United States, black-spot poison ivy dermatitis is relatively rare.

Proteome Changes in biceps femoris Muscle of Iranian One-Humped Camel and Their Effect on Meat Quality Traits

Directory of Open Access Journals (Sweden)

Mohammad-Javad Varidi

2016-01-01

Full Text Available In this study physicochemical and quality traits of biceps femoris and longissimus thoracis muscles of male and female Iranian one-humped camel were determined during 14 days of refrigeration storage. Analysis of variance of the results showed that only shear force and temperature were affected by the gender (p<0.05. Anatomical location of the muscle influenced the meat properties except for drip loss (p<0.05. Also, except for cooking loss, ageing influenced the physicochemical and quality properties of meat; during 14 days of storage, proteolysis resulted in an increase of L* and b* values, drip loss and myofibrillar fragmentation index, and the decrease of a* value, expressed juice, shear force and sarcomere length. Proteome changes (myofi brillar proteins during storage were investigated. Gel analysis revealed that 19 protein spots were signifi cantly changed during 24, 72 and 168 h post-mortem. Fifteen spots were identified by MALDI-TOF/TOF mass spectrometer. Correlation analysis revealed significant correlations of actin, troponin T, capping protein, heat shock proteins (HSP and desmin with physicochemical and quality properties of meat (p<0.05. Actin might be a potential protein marker for colour, tenderness and water-holding capacity, and HSP27 and desmin are good candidate markers for colour and tenderness, respectively.

7 CFR 1421.11 - Spot checks.

Science.gov (United States)

2010-01-01

... 7 Agriculture 10 2010-01-01 2010-01-01 false Spot checks. 1421.11 Section 1421.11 Agriculture... ASSISTANCE LOANS AND LOAN DEFICIENCY PAYMENTS FOR 2008 THROUGH 2012 General § 1421.11 Spot checks. (a) CCC... CCC access to the farm and storage facility as necessary to conduct collateral inspections, or “spot...

21 CFR 886.1435 - Maxwell spot.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Maxwell spot. 886.1435 Section 886.1435 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Diagnostic Devices § 886.1435 Maxwell spot. (a) Identification. A Maxwell spot is an AC...

Newborn screening blood spot analysis in the UK: influence of spot size, punch location and haematocrit.

Science.gov (United States)

Lawson, A J; Bernstone, L; Hall, S K

2016-03-01

In dried blood spot analysis, punch location and variations in applied sample volume and haematocrit can produce different measured concentrations of analytes. We investigated the magnitude of these effects in newborn screening in the UK. Heparinized blood spiked with thyroid stimulating hormone (TSH), phenylalanine, tyrosine, leucine, methionine, octanoyl carnitine (C8), and immunoreactive trypsinogen (IRT) was spotted onto filter paper: (i) at a constant haematocrit of 50% at various volumes, and (ii) at a range of haematocrits using a constant volume. Subpunches (3.2 mm) of the dried blood spots were then analysed. Compared with a central punch from a 50 µL blood spot with 50% haematocrit, 10 µL spots can have significantly lower measured concentrations of all analytes, with decreases of 15% or more observed for leucine, methionine, phenylalanine, and tyrosine. Punching at the edge of a spot can increase measured concentrations up to 35%. Higher haematocrit decreased measured TSH and C8 yet increased amino acids and IRT by 15% compared with 50% haematocrit. Lower haematocrits had the opposite effect, but only with higher concentrations of some analytes. Differences in blood spot size, haematocrit and punch location substantially affect measured concentrations for analytes used in the UK newborn screening programme, and this could affect false positive and negative rates. To minimize analytical bias, these variables should be controlled or adjusted for where possible. © The Author(s) 2015.

Protein change in plant evolution: tracing one thread connecting molecular and phenotypic diversity

Directory of Open Access Journals (Sweden)

Madelaine eBartlett

2013-10-01

Full Text Available Proteins change over the course of evolutionary time. New protein-coding genes and gene families emerge and diversify, ultimately affecting an organism’s phenotype and interactions with its environment. Here we survey the range of structural protein change observed in plants and review the role these changes have had in the evolution of plant form and function. Verified examples tying evolutionary change in protein structure to phenotypic change remain scarce. We will review the existing examples, as well as draw from investigations into domestication, and quantitative trait locus (QTL cloning studies searching for the molecular underpinnings of natural variation. The evolutionary significance of many cloned QTL has not been assessed, but all the examples identified so far have begun to reveal the extent of protein structural diversity tolerated in natural systems. This molecular (and phenotypic diversity could come to represent part of natural selection’s source material in the adaptive evolution of novel traits. Protein structure and function can change in many distinct ways, but the changes we identified in studies of natural diversity and protein evolution were predicted to fall primarily into one of six categories: altered active and binding sites; hypomorphic and hypermorphic alleles; altered protein-protein interactions; altered domain content; altered protein stability; and altered activity as an activator or repressor. Variability was also observed in the evolutionary scale at which particular changes were observed. Some changes were detected at both micro- and macroevolutionary timescales, while others were observed primarily at deep or shallow phylogenetic levels. This variation might be used to determine the trajectory of future investigations in structural molecular evolution.

Controlling total spot power from holographic laser by superimposing a binary phase grating.

Science.gov (United States)

Liu, Xiang; Zhang, Jian; Gan, Yu; Wu, Liying

2011-04-25

By superimposing a tunable binary phase grating with a conventional computer-generated hologram, the total power of multiple holographic 3D spots can be easily controlled by changing the phase depth of grating with high accuracy to a random power value for real-time optical manipulation without extra power loss. Simulation and experiment results indicate that a resolution of 0.002 can be achieved at a lower time cost for normalized total spot power.

Sequence charge decoration dictates coil-globule transition in intrinsically disordered proteins.

Science.gov (United States)

Firman, Taylor; Ghosh, Kingshuk

2018-03-28

We present an analytical theory to compute conformations of heteropolymers-applicable to describe disordered proteins-as a function of temperature and charge sequence. The theory describes coil-globule transition for a given protein sequence when temperature is varied and has been benchmarked against the all-atom Monte Carlo simulation (using CAMPARI) of intrinsically disordered proteins (IDPs). In addition, the model quantitatively shows how subtle alterations of charge placement in the primary sequence-while maintaining the same charge composition-can lead to significant changes in conformation, even as drastic as a coil (swelled above a purely random coil) to globule (collapsed below a random coil) and vice versa. The theory provides insights on how to control (enhance or suppress) these changes by tuning the temperature (or solution condition) and charge decoration. As an application, we predict the distribution of conformations (at room temperature) of all naturally occurring IDPs in the DisProt database and notice significant size variation even among IDPs with a similar composition of positive and negative charges. Based on this, we provide a new diagram-of-states delineating the sequence-conformation relation for proteins in the DisProt database. Next, we study the effect of post-translational modification, e.g., phosphorylation, on IDP conformations. Modifications as little as two-site phosphorylation can significantly alter the size of an IDP with everything else being constant (temperature, salt concentration, etc.). However, not all possible modification sites have the same effect on protein conformations; there are certain "hot spots" that can cause maximal change in conformation. The location of these "hot spots" in the parent sequence can readily be identified by using a sequence charge decoration metric originally introduced by Sawle and Ghosh. The ability of our model to predict conformations (both expanded and collapsed states) of IDPs at a high

Epidemiological Features and Trends of Brown Spot of Pear Disease Based on the Diversity of Pathogen Populations and Climate Change Effects.

Science.gov (United States)

Moragrega, Concepció; Puig, Mireia; Ruz, Lídia; Montesinos, Emilio; Llorente, Isidre

2018-02-01

Brown spot of pear, caused by the fungus Stemphylium vesicarium, is an emerging disease of economic importance in several pear-growing areas in Europe. In recent years, new control strategies combining sanitation practices and fungicide applications according to developed forecasting models have been introduced to manage the disease. However, the pathogenic and saprophytic behavior of this pathogen makes it difficult to manage the disease. In addition, climate change can also result in variations in the severity and geographical distribution of the disease. In this study, ecological and epidemiological aspects of brown spot of pear disease related to inoculum characterization and climate change impact were elucidated. The pathogenic variation in S. vesicarium populations from pear orchards and its relationship to inoculum sources (air samples, leaf debris, and infected host and nonhost tissues) was determined using multivariate analysis. In total, six variables related to infection and disease development on cultivar Conference pear detached leaves of 110 S. vesicarium isolates were analyzed. A high proportion of isolates (42%) were nonpathogenic to pear; 85% of these nonpathogenic isolates were recovered from air samples. Most isolates recovered from lesions (93%) and pseudothecia (83%) were pathogenic to pear. A group of pathogenic isolates rapidly infected cultivar Conference pear leaves resulted in disease increase that followed a monomolecular model, whereas some S. vesicarium isolates required a period of time after inoculation to initiate infection and resulted in disease increase that followed a logistic model. The latter group was mainly composed of isolates recovered from pseudothecia on leaf debris, whereas the former group was mainly composed of isolates recovered from lesions on pear fruit and leaves. The relationship between the source of inoculum and pathogenic/aggressiveness profile was confirmed by principal component analysis. The effect of

Quinone-induced protein handling changes: Implications for major protein handling systems in quinone-mediated toxicity

International Nuclear Information System (INIS)

Xiong, Rui; Siegel, David; Ross, David

2014-01-01

Para-quinones such as 1,4-Benzoquinone (BQ) and menadione (MD) and ortho-quinones including the oxidation products of catecholamines, are derived from xenobiotics as well as endogenous molecules. The effects of quinones on major protein handling systems in cells; the 20/26S proteasome, the ER stress response, autophagy, chaperone proteins and aggresome formation, have not been investigated in a systematic manner. Both BQ and aminochrome (AC) inhibited proteasomal activity and activated the ER stress response and autophagy in rat dopaminergic N27 cells. AC also induced aggresome formation while MD had little effect on any protein handling systems in N27 cells. The effect of NQO1 on quinone induced protein handling changes and toxicity was examined using N27 cells stably transfected with NQO1 to generate an isogenic NQO1-overexpressing line. NQO1 protected against BQ–induced apoptosis but led to a potentiation of AC- and MD-induced apoptosis. Modulation of quinone-induced apoptosis in N27 and NQO1-overexpressing cells correlated only with changes in the ER stress response and not with changes in other protein handling systems. These data suggested that NQO1 modulated the ER stress response to potentiate toxicity of AC and MD, but protected against BQ toxicity. We further demonstrated that NQO1 mediated reduction to unstable hydroquinones and subsequent redox cycling was important for the activation of the ER stress response and toxicity for both AC and MD. In summary, our data demonstrate that quinone-specific changes in protein handling are evident in N27 cells and the induction of the ER stress response is associated with quinone-mediated toxicity. - Highlights: • Unstable hydroquinones contributed to quinone-induced ER stress and toxicity

Quinone-induced protein handling changes: Implications for major protein handling systems in quinone-mediated toxicity

Energy Technology Data Exchange (ETDEWEB)

Xiong, Rui; Siegel, David; Ross, David, E-mail: david.ross@ucdenver.edu

2014-10-15

Para-quinones such as 1,4-Benzoquinone (BQ) and menadione (MD) and ortho-quinones including the oxidation products of catecholamines, are derived from xenobiotics as well as endogenous molecules. The effects of quinones on major protein handling systems in cells; the 20/26S proteasome, the ER stress response, autophagy, chaperone proteins and aggresome formation, have not been investigated in a systematic manner. Both BQ and aminochrome (AC) inhibited proteasomal activity and activated the ER stress response and autophagy in rat dopaminergic N27 cells. AC also induced aggresome formation while MD had little effect on any protein handling systems in N27 cells. The effect of NQO1 on quinone induced protein handling changes and toxicity was examined using N27 cells stably transfected with NQO1 to generate an isogenic NQO1-overexpressing line. NQO1 protected against BQ–induced apoptosis but led to a potentiation of AC- and MD-induced apoptosis. Modulation of quinone-induced apoptosis in N27 and NQO1-overexpressing cells correlated only with changes in the ER stress response and not with changes in other protein handling systems. These data suggested that NQO1 modulated the ER stress response to potentiate toxicity of AC and MD, but protected against BQ toxicity. We further demonstrated that NQO1 mediated reduction to unstable hydroquinones and subsequent redox cycling was important for the activation of the ER stress response and toxicity for both AC and MD. In summary, our data demonstrate that quinone-specific changes in protein handling are evident in N27 cells and the induction of the ER stress response is associated with quinone-mediated toxicity. - Highlights: • Unstable hydroquinones contributed to quinone-induced ER stress and toxicity.

Plasmonic electromagnetic hot spots temporally addressed by photoinduced molecular displacement.

Energy Technology Data Exchange (ETDEWEB)

Juan, M. L.; Plain, J.; Bachelot, R.; Vial, A.; Royer, P.; Gray, S. K.; Montgomery, J. M.; Wiederrecht, G. P.; Univ. de Technologie de Troyes

2009-04-23

We report the observation of temporally varying electromagnetic hot spots in plasmonic nanostructures. Changes in the field amplitude, position, and spatial features are induced by embedding plasmonic silver nanorods in the photoresponsive azo-polymer. This polymer undergoes cis?trans isomerization and wormlike transport within resonant optical fields, producing a time-varying local dielectric environment that alters the locations where electromagnetic hot spots are produced. Finite-difference time-domain and Monte Carlo simulations that model the induced field and corresponding material response are presented to aid in the interpretation of the experimental results. Evidence for propagating plasmons induced at the ends of the rods is also presented.

Optimised 'on demand' protein arraying from DNA by cell free expression with the 'DNA to Protein Array' (DAPA) technology.

Science.gov (United States)

Schmidt, Ronny; Cook, Elizabeth A; Kastelic, Damjana; Taussig, Michael J; Stoevesandt, Oda

2013-08-02

We have previously described a protein arraying process based on cell free expression from DNA template arrays (DNA Array to Protein Array, DAPA). Here, we have investigated the influence of different array support coatings (Ni-NTA, Epoxy, 3D-Epoxy and Polyethylene glycol methacrylate (PEGMA)). Their optimal combination yields an increased amount of detected protein and an optimised spot morphology on the resulting protein array compared to the previously published protocol. The specificity of protein capture was improved using a tag-specific capture antibody on a protein repellent surface coating. The conditions for protein expression were optimised to yield the maximum amount of protein or the best detection results using specific monoclonal antibodies or a scaffold binder against the expressed targets. The optimised DAPA system was able to increase by threefold the expression of a representative model protein while conserving recognition by a specific antibody. The amount of expressed protein in DAPA was comparable to those of classically spotted protein arrays. Reaction conditions can be tailored to suit the application of interest. DAPA represents a cost effective, easy and convenient way of producing protein arrays on demand. The reported work is expected to facilitate the application of DAPA for personalized medicine and screening purposes. Copyright © 2013 Elsevier B.V. All rights reserved.

The fractal nature of vacuum arc cathode spots

International Nuclear Information System (INIS)

Anders, Andre

2005-01-01

Cathode spot phenomena show many features of fractals, for example self-similar patterns in the emitted light and arc erosion traces. Although there have been hints on the fractal nature of cathode spots in the literature, the fractal approach to spot interpretation is underutilized. In this work, a brief review of spot properties is given, touching the differences between spot type 1 (on cathodes surfaces with dielectric layers) and spot type 2 (on metallic, clean surfaces) as well as the known spot fragment or cell structure. The basic properties of self-similarity, power laws, random colored noise, and fractals are introduced. Several points of evidence for the fractal nature of spots are provided. Specifically power laws are identified as signature of fractal properties, such as spectral power of noisy arc parameters (ion current, arc voltage, etc) obtained by fast Fourier transform. It is shown that fractal properties can be observed down to the cutoff by measurement resolution or occurrence of elementary steps in physical processes. Random walk models of cathode spot motion are well established: they go asymptotically to Brownian motion for infinitesimal step width. The power spectrum of the arc voltage noise falls as 1/f 2 , where f is frequency, supporting a fractal spot model associated with Brownian motion

Establishment of a high-resolution 2-D reference map of human spermatozoal proteins from 12 fertile sperm-bank donors.

Science.gov (United States)

Li, Ling-Wei; Fan, Li-Qing; Zhu, Wen-Bing; Nien, Hong-Chuan; Sun, Bo-Lan; Luo, Ke-Li; Liao, Ting-Ting; Tang, Le; Lu, Guang-Xiu

2007-05-01

To extend the analysis of the proteome of human spermatozoa and establish a 2-D gel electrophoresis (2-DE) reference map of human spermatozoal proteins in a pH range of 3.5-9.0. In order to reveal more protein spots, immobilized pH gradient strips (24 cm) of broad range of pH 3-10 and the narrower range of pH 6-9, as well as different overlapping narrow range pH immobilized pH gradient (IPG) strips, including 3.5-4.5, 4.0-5.0, 4.5-5.5, 5.0-6.0 and 5.5-6.7, were used. After 2-DE, several visually identical spots between the different pH range 2-D gel pairs were cut from the gels and confirmed by mass spectrometry and used as landmarks for computer analysis. The 2-D reference map with pH value from 3.5 to 9.0 was synthesized by using the ImageMaster analysis software. The overlapping spots were excluded, so that every spot was counted only once. A total of 3872 different protein spots were identified from the reference map, an approximately 3-fold increase compared to the broad range pH 3-10 IPG strip (1306 spots). The present 2-D pattern is a high resolution 2-D reference map for human fertile spermatozoal protein spots. A comprehensive knowledge of the protein composition of human spermatozoa is very meaningful in studying dysregulation of male fertility.

Modeling co-occurrence of northern spotted and barred owls: accounting for detection probability differences

Science.gov (United States)

Bailey, Larissa L.; Reid, Janice A.; Forsman, Eric D.; Nichols, James D.

2009-01-01

Barred owls (Strix varia) have recently expanded their range and now encompass the entire range of the northern spotted owl (Strix occidentalis caurina). This expansion has led to two important issues of concern for management of northern spotted owls: (1) possible competitive interactions between the two species that could contribute to population declines of northern spotted owls, and (2) possible changes in vocalization behavior and detection probabilities of northern spotted owls induced by presence of barred owls. We used a two-species occupancy model to investigate whether there was evidence of competitive exclusion between the two species at study locations in Oregon, USA. We simultaneously estimated detection probabilities for both species and determined if the presence of one species influenced the detection of the other species. Model selection results and associated parameter estimates provided no evidence that barred owls excluded spotted owls from territories. We found strong evidence that detection probabilities differed for the two species, with higher probabilities for northern spotted owls that are the object of current surveys. Non-detection of barred owls is very common in surveys for northern spotted owls, and detection of both owl species was negatively influenced by the presence of the congeneric species. Our results suggest that analyses directed at hypotheses of barred owl effects on demographic or occupancy vital rates of northern spotted owls need to deal adequately with imperfect and variable detection probabilities for both species.

Protein and metabolite composition of xylem sap from field-grown soybeans (Glycine max).

Science.gov (United States)

Krishnan, Hari B; Natarajan, Savithiry S; Bennett, John O; Sicher, Richard C

2011-05-01

The xylem, in addition to transporting water, nutrients and metabolites, is also involved in long-distance signaling in response to pathogens, symbionts and environmental stresses. Xylem sap has been shown to contain a number of proteins including metabolic enzymes, stress-related proteins, signal transduction proteins and putative transcription factors. Previous studies on xylem sap have mostly utilized plants grown in controlled environmental chambers. However, plants in the field are subjected to high light and to environmental stress that is not normally found in growth chambers. In this study, we have examined the protein and metabolite composition of xylem sap from field-grown cultivated soybean plants. One-dimensional gel electrophoresis of xylem sap from determinate, indeterminate, nodulating and non-nodulating soybean cultivars revealed similar protein profiles consisting of about 8-10 prominent polypeptides. Two-dimensional gel electrophoresis of soybean xylem sap resulted in the visualization of about 60 distinct protein spots. A total of 38 protein spots were identified using MALDI-TOF MS and LC-MS/MS. The most abundant proteins present in the xylem sap were identified as 31 and 28 kDa vegetative storage proteins. In addition, several proteins that are conserved among different plant species were also identified. Diurnal changes in the metabolite profile of xylem sap collected during a 24-h cycle revealed that asparagine and aspartate were the two predominant amino acids irrespective of the time collected. Pinitol (D-3-O-methyl-chiro-inositol) was the most abundant carbohydrate present. The possible roles of xylem sap proteins and metabolites as nutrient reserves for sink tissue and as an indicator of biotic stress are also discussed.

Topographic hot spot before descemet stripping automated endothelial keratoplasty is associated with postoperative hyperopic shift.

Science.gov (United States)

Shimizu, Tsutomu; Yamaguchi, Takefumi; Satake, Yoshiyuki; Shimazaki, Jun

2015-03-01

The aim of this study was to investigate topographic "hot spots" on the anterior corneal surface before Descemet stripping automated endothelial keratoplasty (DSAEK) and their effects on postoperative visual acuity and hyperopic shift. Twenty-seven eyes of 27 patients with bullous keratopathy, who underwent DSAEK were studied. We defined a hot spot as a focal area with relatively high refractive power on the anterior corneal surface in eyes with bullous keratopathy. Refractive spherical equivalent, keratometric value, and corneal topography were retrospectively evaluated using anterior segment optical coherence tomography (AS-OCT). Hot spots were identified in 11 eyes (42.3%) before DSAEK and disappeared in 9 eyes of these eyes (81.8%) at 6 months after DSAEK. AS-OCT revealed focal epithelial thickening in the same areas as the hot spots. There was no significant difference in the postoperative visual acuity between eyes with and without hot spots (P > 0.05). The keratometric value of the anterior corneal surface significantly flattened from 45.7 ± 2.7 diopters (D) before DSAEK to 44.2 ± 2.7 D 1 month after DSAEK in eyes with hot spots (P = 0.01), whereas in eyes without hot spots, there were no significant differences in the keratometric values before and after DSAEK. At 6 months, the refractive change was +1.1 ± 1.3 D in eyes with hot spots and -0.2 ± 0.6 D in eyes without hot spots (P = 0.034). In eyes with focal epithelial thickening, topographic hot spots on the anterior corneal surface were observed using AS-OCT. The hot spots disappeared after DSAEK and had no influence on the postoperative visual acuity.

Laser reflection spot as a pattern in a diamond coating – a microscopic study

Directory of Open Access Journals (Sweden)

GORDANA S. RISTIĆ

2009-07-01

Full Text Available Diamond coatings were deposited by the synchronous and coupled action of a hot filament CVD method and a pulsed CO2 laser in spectro-absorbing and spectro-non-absorbing diamond precursor atmospheres. The obtained coatings were structured/patterned, i.e., they were comprised of uncovered, bare locations. An extra effect observed only in the spectro-active diamond precursor atmosphere was the creation of another laser spot in the coating – a reflection spot. In order to establish the practical usability of the latter one, extensive microscopic investigations were performed with consideration of the morphology changes in the spot of the direct laser beam. Normal incidence SEM images of this spot showed a smooth surface, without any pulse radiation damage. AFM imaging revealed the actual surface condition and gave precise data on the surface characteristics.

The Influence of spot size on the expansion dynamics of nanosecond-laser-produced copper plasmas in atmosphere

International Nuclear Information System (INIS)

Li, Xingwen; Wei, Wenfu; Wu, Jian; Jia, Shenli; Qiu, Aici

2013-01-01

Laser produced copper plasmas of different spot sizes in air were investigated using fast photography and optical emission spectroscopy (OES). The laser energy was 33 mJ. There were dramatic changes in the plasma plume expansion into the ambient air when spot sizes changed from ∼0.1 mm to ∼0.6 mm. A stream-like structure and a hemispherical structure were, respectively, observed. It appeared that the same spot size resulted in similar expansion dynamics no matter whether the target was located in the front of or behind the focal point, although laser-induced air breakdown sometimes occurred in the latter case. Plasma plume front positions agree well with the classic blast wave model for the large spot-size cases, while an unexpected stagnation of ∼80 ns occurred after the laser pulse ends for the small spot size cases. This stagnation can be understood in terms of the evolution of enhanced plasma shielding effects near the plasma front. Axial distributions of plasma components by OES revealed a good confinement effect. Electron number densities were estimated and interpreted using the recorded Intensified Charge Coupled Device (ICCD) images.

Proteomic analysis of seed storage proteins in wild rice species of the Oryza genus.

Science.gov (United States)

Jiang, Chunmiao; Cheng, Zaiquan; Zhang, Cheng; Yu, Tengqiong; Zhong, Qiaofang; Shen, J Qingxi; Huang, Xingqi

2014-01-01

The total protein contents of rice seeds are significantly higher in the three wild rice species (Oryza rufipogon Grill., Oryza officinalis Wall. and Oryza meyeriana Baill.) than in the cultivated rice (Oryza sativa L.). However, there is still no report regarding a systematic proteomic analysis of seed proteins in the wild rice species. Also, the relationship between the contents of seed total proteins and rice nutritional quality has not been thoroughly investigated. The total seed protein contents, especially the glutelin contents, of the three wild rice species were higher than those of the two cultivated rice materials. Based on the protein banding patterns of SDS-PAGE, O. rufipogon was similar to the two cultivated rice materials, followed by O. officinalis, while O. meyeriana exhibited notable differences. Interestingly, O. meyeriana had high contents of glutelin and low contents of prolamine, and lacked 26 kDa globulin band and appeared a new 28 kDa protein band. However, for O. officinali a 16 kDa protein band was absent and a row of unique 32 kDa proteins appeared. In addition, we found that 13 kDa prolamine band disappeared while special 14 kDa and 12 kDa protein bands were present in O. officinalis. Two-dimensional gel electrophoresis (2-DE) analysis revealed remarkable differences in protein profiles of the wild rice species and the two cultivated rice materials. Also, the numbers of detected protein spots of the three wild rice species were significantly higher than those of two cultivated rice. A total of 35 differential protein spots were found for glutelin acidic subunits, glutelin precursors and glutelin basic subunits in wild rice species. Among those, 18 protein spots were specific and 17 major spots were elevated. Six differential protein spots for glutelin acidic subunits were identified, including a glutelin type-A 2 precursor and five hypothetical proteins. This was the first report on proteomic analysis of the three wild rice species

Laser Spot Detection Based on Reaction Diffusion

OpenAIRE

Alejandro Vázquez-Otero; Danila Khikhlukha; J. M. Solano-Altamirano; Raquel Dormido; Natividad Duro

2016-01-01

Center-location of a laser spot is a problem of interest when the laser is used for processing and performing measurements. Measurement quality depends on correctly determining the location of the laser spot. Hence, improving and proposing algorithms for the correct location of the spots are fundamental issues in laser-based measurements. In this paper we introduce a Reaction Diffusion (RD) system as the main computational framework for robustly finding laser spot centers. The method presente...

7 CFR 28.415 - Low Middling Light Spotted Color.

Science.gov (United States)

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Low Middling Light Spotted Color. 28.415 Section 28... Spotted Color. Low Middling Light Spotted Color is color which in spot or color, or both, is between Low Middling Color and Low Middling Spotted Color. ...

Spotted alfalfa aphid, Therioaphis trifolii (Monell) (Hemiptera: Aphididae): Pest on alfalfa in Serbia

OpenAIRE

Jovičić, Ivana; Radonjić, Anđa; Petrović-Obradović, Olivera

2017-01-01

Spotted alfalfa aphid Therioaphis trifolii (Monell) (Hemiptera, Aphididae) is one of the most important alfalfa pest on the world. Also, it is the most abundant alfalfa aphid in Serbia. This aphid cause damage to alfalfa directly by feeding and indirectly by vectoring plant-pathogenic viruses. Some notes of morphology, host plants, damage, biology, vector role and distribution of spotted alfalfa aphid are given. Abundance of this aphid on alfalfa, influence of climates changes on its abundanc...

Facial skin photo-aging and development of hyperpigmented spots from children to middle-aged Japanese woman.

Science.gov (United States)

Takahashi, Y; Fukushima, Y; Kondo, K; Ichihashi, M

2017-11-01

Facial skin hyperpigmention caused by chronic sun exposure is a major skin complaint, however, its characteristics and influential factors are still limitedly known. A cross-sectional survey in healthy Japanese women aged from 6 to 62 years (n=169) was conducted using a facial image analyzer VISIA ™ for knowing onset age of hyperpigmented spot formation, its chronological changes, and influence of environmental factors. UV Pigmented Spot (PS) Score was positively correlated with age (R=.487, P=.000). Hyperpigmented spots appeared first around 18 years old in most subjects, and PS score remarkably increased at 20s then gradually increased by ages. The subjects with Skin Type I, one of the three grades of Japanese Skin Type (JST), whose melanin formation is genetically lower, showed higher PS score. A woman aged 31 years was subjected a weekly VISIA measurement for 2 years, and found no changes in the number, place, size and intensity of the pigment spots in this duration. Hyperpigmented spots developed in women over 20 years of age due to chronic sun exposure without sun protection during childhood and adolescent and it was stable afterwards, whose intensity was influenced by age and skin type. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

PyTranSpot: A tool for multiband light curve modeling of planetary transits and stellar spots

Science.gov (United States)

Juvan, Ines G.; Lendl, M.; Cubillos, P. E.; Fossati, L.; Tregloan-Reed, J.; Lammer, H.; Guenther, E. W.; Hanslmeier, A.

2018-02-01

Several studies have shown that stellar activity features, such as occulted and non-occulted starspots, can affect the measurement of transit parameters biasing studies of transit timing variations and transmission spectra. We present PyTranSpot, which we designed to model multiband transit light curves showing starspot anomalies, inferring both transit and spot parameters. The code follows a pixellation approach to model the star with its corresponding limb darkening, spots, and transiting planet on a two dimensional Cartesian coordinate grid. We combine PyTranSpot with a Markov chain Monte Carlo framework to study and derive exoplanet transmission spectra, which provides statistically robust values for the physical properties and uncertainties of a transiting star-planet system. We validate PyTranSpot's performance by analyzing eleven synthetic light curves of four different star-planet systems and 20 transit light curves of the well-studied WASP-41b system. We also investigate the impact of starspots on transit parameters and derive wavelength dependent transit depth values for WASP-41b covering a range of 6200-9200 Å, indicating a flat transmission spectrum.

Comparative proteomic analysis of Xanthomonas citri ssp. citri periplasmic proteins reveals changes in cellular envelope metabolism during in vitro pathogenicity induction.

Science.gov (United States)

Artier, Juliana; da Silva Zandonadi, Flávia; de Souza Carvalho, Flávia Maria; Pauletti, Bianca Alves; Leme, Adriana Franco Paes; Carnielli, Carolina Moretto; Selistre-de-Araujo, Heloisa Sobreiro; Bertolini, Maria Célia; Ferro, Jesus Aparecido; Belasque Júnior, José; de Oliveira, Julio Cezar Franco; Novo-Mansur, Maria Teresa Marques

2018-01-01

Citrus canker is a plant disease caused by Gram-negative bacteria from the genus Xanthomonas. The most virulent species is Xanthomonas citri ssp. citri (XAC), which attacks a wide range of citrus hosts. Differential proteomic analysis of the periplasm-enriched fraction was performed for XAC cells grown in pathogenicity-inducing (XAM-M) and pathogenicity-non-inducing (nutrient broth) media using two-dimensional electrophoresis combined with liquid chromatography-tandem mass spectrometry. Amongst the 40 proteins identified, transglycosylase was detected in a highly abundant spot in XAC cells grown under inducing condition. Additional up-regulated proteins related to cellular envelope metabolism included glucose-1-phosphate thymidylyltransferase, dTDP-4-dehydrorhamnose-3,5-epimerase and peptidyl-prolyl cis-trans-isomerase. Phosphoglucomutase and superoxide dismutase proteins, known to be involved in pathogenicity in other Xanthomonas species or organisms, were also detected. Western blot and quantitative real-time polymerase chain reaction analyses for transglycosylase and superoxide dismutase confirmed that these proteins were up-regulated under inducing condition, consistent with the proteomic results. Multiple spots for the 60-kDa chaperonin and glyceraldehyde-3-phosphate dehydrogenase were identified, suggesting the presence of post-translational modifications. We propose that substantial alterations in cellular envelope metabolism occur during the XAC infectious process, which are related to several aspects, from defence against reactive oxygen species to exopolysaccharide synthesis. Our results provide new candidates for virulence-related proteins, whose abundance correlates with the induction of pathogenicity and virulence genes, such as hrpD6, hrpG, hrpB7, hpa1 and hrpX. The results present new potential targets against XAC to be investigated in further functional studies. © 2016 BSPP AND JOHN WILEY & SONS LTD.

Rocky Mountain spotted fever in Argentina.

Science.gov (United States)

Paddock, Christopher D; Fernandez, Susana; Echenique, Gustavo A; Sumner, John W; Reeves, Will K; Zaki, Sherif R; Remondegui, Carlos E

2008-04-01

We describe the first molecular confirmation of Rickettsia rickettsii, the cause of Rocky Mountain spotted fever (RMSF), from a tick vector, Amblyomma cajennense, and from a cluster of fatal spotted fever cases in Argentina. Questing A. cajennense ticks were collected at or near sites of presumed or confirmed cases of spotted fever rickettsiosis in Jujuy Province and evaluated by polymerase chain reaction assays for spotted fever group rickettsiae. DNA of R. rickettsii was amplified from a pool of A. cajennense ticks and from tissues of one of four patients who died during 2003-2004 after illnesses characterized by high fever, severe headache, myalgias, and petechial rash. The diagnosis of spotted fever rickettsiosis was confirmed in the other patients by indirect immunofluorescence antibody and immunohistochemical staining techniques. These findings show the existence of RMSF in Argentina and emphasize the need for clinicians throughout the Americas to consider RMSF in patients with febrile rash illnesses.

Crack imaging by pulsed laser spot thermography

International Nuclear Information System (INIS)

Li, T; Almond, D P; Rees, D A S; Weekes, B

2010-01-01

A surface crack close to a spot heated by a laser beam impedes lateral heat flow and produces alterations to the shape of the thermal image of the spot that can be monitored by thermography. A full 3D simulation has been developed to simulate heat flow from a laser heated spot in the proximity of a crack. The modelling provided an understanding of the ways that different parameters affect the thermal images of laser heated spots. It also assisted in the development of an efficient image processing strategy for extracting the scanned cracks. Experimental results show that scanning pulsed laser spot thermography has considerable potential as a remote, non-contact crack imaging technique.

Spot Welding of Honeycomb Structures

Science.gov (United States)

Cohal, V.

2017-08-01

Honeycomb structures are used to prepare meals water jet cutting machines for textile. These honeycomb structures are made of stainless steel sheet thickness of 0.1-0.2 mm. Corrugated sheet metal strips are between two gears with special tooth profile. Hexagonal cells for obtaining these strips are welded points between them. Spot welding device is three electrodes in the upper part, which carries three welding points across the width of the strip of corrugated sheet metal. Spot welding device filled with press and advance mechanisms. The paper presents the values of the regime for spot welding.

7 CFR 28.412 - Strict Middling Light Spotted Color.

Science.gov (United States)

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Light Spotted Color. 28.412 Section 28... Light Spotted Color. Strict Middling Light Spotted Color is color which in spot or color, or both, is between Strict Middling Color and Strict Middling Spotted Color. ...

7 CFR 28.411 - Good Middling Light Spotted Color.

Science.gov (United States)

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Good Middling Light Spotted Color. 28.411 Section 28... Light Spotted Color. Good Middling Light Spotted Color is color which in spot or color, or both, is between Good Middling Color and Good Middling Spotted Color. ...

Persistence at distributional edges: Columbia spotted frog habitat in the arid Great Basin, USA.

Science.gov (United States)

Arkle, Robert S; Pilliod, David S

2015-09-01

A common challenge in the conservation of broadly distributed, yet imperiled species is understanding which factors facilitate persistence at distributional edges, locations where populations are often vulnerable to extirpation due to changes in climate, land use, or distributions of other species. For Columbia spotted frogs (Rana luteiventris) in the Great Basin (USA), a genetically distinct population segment of conservation concern, we approached this problem by examining (1) landscape-scale habitat availability and distribution, (2) water body-scale habitat associations, and (3) resource management-identified threats to persistence. We found that areas with perennial aquatic habitat and suitable climate are extremely limited in the southern portion of the species' range. Within these suitable areas, native and non-native predators (trout and American bullfrogs [Lithobates catesbeianus]) are widespread and may further limit habitat availability in upper- and lower-elevation areas, respectively. At the water body scale, spotted frog occupancy was associated with deeper sites containing abundant emergent vegetation and nontrout fish species. Streams with American beaver (Castor canadensis) frequently had these structural characteristics and were significantly more likely to be occupied than ponds, lakes, streams without beaver, or streams with inactive beaver ponds, highlighting the importance of active manipulation of stream environments by beaver. Native and non-native trout reduced the likelihood of spotted frog occupancy, especially where emergent vegetation cover was sparse. Intensive livestock grazing, low aquatic connectivity, and ephemeral hydroperiods were also negatively associated with spotted frog occupancy. We conclude that persistence of this species at the arid end of its range has been largely facilitated by habitat stability (i.e., permanent hydroperiod), connectivity, predator-free refugia, and a commensalistic interaction with an ecosystem

Detection of spatial hot spots and variation for the neon flying squid Ommastrephes bartramii resources in the northwest Pacific Ocean

Science.gov (United States)

Feng, Yongjiu; Chen, Xinjun; Liu, Yan

2017-07-01

With the increasing effects of global climate change and fishing activities, the spatial distribution of the neon flying squid ( Ommastrephes bartramii) is changing in the traditional fishing ground of 150°-160°E and 38°-45°N in the northwest Pacific Ocean. This research aims to identify the spatial hot and cold spots (i.e. spatial clusters) of O. bartramii to reveal its spatial structure using commercial fishery data from 2007 to 2010 collected by Chinese mainland squid-jigging fleets. A relatively strongly-clustered distribution for O. bartramii was observed using an exploratory spatial data analysis (ESDA) method. The results show two hot spots and one cold spot in 2007 while only one hot and one cold spots were identified each year from 2008 to 2010. The hot and cold spots in 2007 occupied 8.2% and 5.6% of the study area, respectively; these percentages for hot and cold spot areas were 5.8% and 3.1% in 2008, 10.2% and 2.9% in 2009, and 16.4% and 11.9% in 2010, respectively. Nearly half (>45%) of the squid from 2007 to 2009 reported by Chinese fleets were caught in hot spot areas while this percentage reached its peak at 68.8% in 2010, indicating that the hot spot areas are central fishing grounds. A further change analysis shows the area centered at 156°E/43.5°N was persistent as a hot spot over the whole period from 2007 to 2010. Furthermore, the hot spots were mainly identified in areas with sea surface temperature (SST) in the range of 15-20°C around warm Kuroshio Currents as well as with the chlorophyll- a (chl- a) concentration above 0.3 mg/m3. The outcome of this research improves our understanding of spatiotemporal hotspots and its variation for O. bartramii and is useful for sustainable exploitation, assessment, and management of this squid.

Plasmodium falciparum HRP2 ELISA for analysis of dried blood spot samples in rural Zambia.

Science.gov (United States)

Gibson, Lauren E; Markwalter, Christine F; Kimmel, Danielle W; Mudenda, Lwiindi; Mbambara, Saidon; Thuma, Philip E; Wright, David W

2017-08-23

Dried blood spots are commonly used for sample collection in clinical and non-clinical settings. This method is simple, and biomolecules in the samples remain stable for months at room temperature. In the field, blood samples for the study and diagnosis of malaria are often collected on dried blood spot cards, so development of a biomarker extraction and analysis method is needed. A simple extraction procedure for the malarial biomarker Plasmodium falciparum histidine-rich protein 2 (HRP2) from dried blood spots was optimized to achieve maximum extraction efficiency. This method was used to assess the stability of HRP2 in dried blood spots. Furthermore, 328 patient samples made available from rural Zambia were analysed for HRP2 using the developed method. These samples were collected at the initial administration of artemisinin-based combination therapy and at several points following treatment. An average extraction efficiency of 70% HRP2 with a low picomolar detection limit was achieved. In specific storage conditions HRP2 was found to be stable in dried blood spots for at least 6 months. Analysis of patient samples showed the method to have a sensitivity of 94% and a specificity of 89% when compared with microscopy, and trends in HRP2 clearance after treatment were observed. The dried blood spot ELISA for HRP2 was found to be sensitive, specific and accurate. The method was effectively used to assess biomarker clearance characteristics in patient samples, which prove it to be ideal for gaining further insight into the disease and epidemiological applications.

Effect of ambient humidity on the rate at which blood spots dry and the size of the spot produced.

Science.gov (United States)

Denniff, Philip; Woodford, Lynsey; Spooner, Neil

2013-08-01

For shipping and storage, dried blood spot (DBS) samples must be sufficiently dry to protect the integrity of the sample. When the blood is spotted the humidity has the potential to affect the size of the spot created and the speed at which it dries. The area of DBS produced on three types of substrates were not affected by the humidity under which they were generated. DBS samples reached a steady moisture content 150 min after spotting and 90 min for humidities less than 60% relative humidity. All packaging materials examined provided some degree of protection from external extreme conditions. However, none of the packaging examined provided a total moisture barrier to extreme environmental conditions. Humidity was shown not to affect the spot area and DBS samples were ready for shipping and storage 2 h after spotting. The packing solutions examined all provided good protection from external high humidity conditions.

PROTEIN EXTRACTION OF MAIZE (ZEA MAYS FOR PROTEOMIC 2 – DE ANALYSIS

Directory of Open Access Journals (Sweden)

Edita Gregová

2014-12-01

Full Text Available A strength of two-dimensional polyacrylamide gel electrophoresis (2D PAGE is its ability to resolve and investigate the abundance of several thousand proteins in a single sample. Two different extraction procedures for two-dimensional electrophoresis of plant proteins are compared in this work. Phenol-based extraction methods have been mainly used to extract proteins from different organs or tissues on many species. We wanted to determine which of these protocols was optimal for starch plants in order to achieve both efficient protein extraction and high spot resolution on 2-D gels. The phenol-based protocol was superior to the sodium phosphatase methods, showing larger protein yields and greater spot resolution on 2-D gels.

Unblinding the dark matter blind spots

International Nuclear Information System (INIS)

Han, Tao; Kling, Felix

2017-01-01

The dark matter (DM) blind spots in the Minimal Supersymmetric Standard Model (MSSM) refer to the parameter regions where the couplings of the DM particles to the Z-boson or the Higgs boson are almost zero, leading to vanishingly small signals for the DM direct detections. In this paper, we carry out comprehensive analyses for the DM searches under the blind-spot scenarios in MSSM. Guided by the requirement of acceptable DM relic abundance, we explore the complementary coverage for the theory parameters at the LHC, the projection for the future underground DM direct searches, and the indirect searches from the relic DM annihilation into photons and neutrinos. We find that (i) the spin-independent (SI) blind spots may be rescued by the spin-dependent (SD) direct detection in the future underground experiments, and possibly by the indirect DM detections from IceCube and SuperK neutrino experiments; (ii) the detection of gamma rays from Fermi-LAT may not reach the desirable sensitivity for searching for the DM blind-spot regions; (iii) the SUSY searches at the LHC will substantially extend the discovery region for the blind-spot parameters. As a result, the dark matter blind spots thus may be unblinded with the collective efforts in future DM searches.

Evaluation of actual vs expected photodynamic therapy spot size.

Science.gov (United States)

Ranchod, Tushar M; Brucker, Alexander J; Liu, Chengcheng; Cukras, Catherine A; Hopkins, Tim B; Ying, Gui-Shuang

2009-05-01

To determine the accuracy of the photodynamic therapy (PDT) laser spot size on the retina as generated by 2 Food and Drug Administration (FDA)-approved lasers. Prospective observational case series. Fundus photographs were taken of 1 eye of each of 10 subjects with the WinStation 4000 fundus photography system (OIS; Ophthalmic Imaging Systems, Sacramento, California, USA); disc size was calculated using OIS software. Slit-lamp photographs were taken of the PDT laser spot focused on the retina adjacent to the optic disc, using various spot sizes in combination with 3 different contact lenses and 2 different lasers. Spot size at the retina was determined by measuring the ratio of disc diameter to spot diameter in Adobe Photoshop (San Jose, California, USA) and applying this ratio to the OIS disc measurements. Spot size at the retina averaged 87% of expected spot size for the Coherent Opal laser (Coherent Inc, Santa Clara, California, USA) and 104% of expected spot size for the Zeiss Visulas laser (Carl Zeiss Meditec Inc, Dublin, California, USA)(P = .002). Multivariate analysis demonstrated that percentage of expected spot size decreased with larger spot diameter (P = .01 for Coherent laser; P = .02 for Zeiss laser). PDT spot size at the retina appears to be consistently smaller than expected for the Coherent laser while the spot size was consistently within 10% of expected size for the Zeiss laser. The deviation from expected size increased with larger spot size using the Coherent laser.

Laser Spot Detection Based on Reaction Diffusion.

Science.gov (United States)

Vázquez-Otero, Alejandro; Khikhlukha, Danila; Solano-Altamirano, J M; Dormido, Raquel; Duro, Natividad

2016-03-01

Center-location of a laser spot is a problem of interest when the laser is used for processing and performing measurements. Measurement quality depends on correctly determining the location of the laser spot. Hence, improving and proposing algorithms for the correct location of the spots are fundamental issues in laser-based measurements. In this paper we introduce a Reaction Diffusion (RD) system as the main computational framework for robustly finding laser spot centers. The method presented is compared with a conventional approach for locating laser spots, and the experimental results indicate that RD-based computation generates reliable and precise solutions. These results confirm the flexibility of the new computational paradigm based on RD systems for addressing problems that can be reduced to a set of geometric operations.

Insights from ecological niche modeling on the taxonomic distinction and niche differentiation between the black-spotted and red-spotted tokay geckoes (Gekko gecko)

OpenAIRE

Zhang, Yueyun; Chen, Chongtao; Li, Li; Zhao, Chengjian; Chen, Weicai; Huang, Yong

2014-01-01

The black-spotted tokay and the red-spotted tokay are morphologically distinct and have largely allopatric distributions. The black-spotted tokay is characterized by a small body size and dark skin with sundry spots, while the red-spotted tokay has a relatively large body size and red spots. Based on morphological, karyotypic, genetic, and distribution differences, recent studies suggested their species status; however, their classifications remain controversial, and additional data such as e...

Small laser spot versus standard laser spot photodynamic therapy for idiopathic choroidal neovascularization: a randomized controlled study.

Science.gov (United States)

Li, Xiao-xin; Tao, Yong

2012-12-01

Idiopathic choroidal neovascularization (ICNV) affects young patients and thus may have a significant impact on vision and life quality over a patient's lifespan. This study was designed to compare the visual outcome and retinal pigment epithelium (RPE) damage after photodynamic therapy (PDT) with small laser spot and PDT with standard laser spot for idiopathic choroidal neovascularization (ICNV). This was a randomized controlled study. Fifty-two patients with ICNV were enrolled and randomly divided into a study group (small laser spot PDT, n = 27) and a control group (standard laser spot PDT, n = 25). Best corrected visual acuity (BCVA), optic coherence tomography (OCT) and fluorescein angiography (FA) findings were the main measurements. The patients were followed up 1 week, 1, 3, 6, 9 months and 1 year after PDT. BCVA improvement was statistically significantly higher in the study group than the control group at 6-month ((25.53 ± 15.01) letters vs. (14.71 ± 11.66) letters, P = 0.025) and 9-month follow-ups ((27.53 ± 17.78) letters vs. (15.59 ± 12.21) letters, P = 0.039). At 3- and 6-month follow-ups, the quadrants of RPE damage between the two groups varied significantly (P laser spot PDT group than in the standard laser spot PDT group for ICNV.

Application of a Label-Free Immunosensor for White Spot Syndrome Virus (WSSV) in Shrimp Cultivation Water.

Science.gov (United States)

Waiyapoka, Thanyaporn; Deachamag, Panchalika; Chotigeat, Wilaiwan; Bunsanong, Nittaya; Kanatharana, Proespichaya; Thavarungkul, Panote; Loyprasert-Thananimit, Suchera

2015-10-01

White spot syndrome virus (WSSV) is a major pathogen affecting the shrimp industry worldwide. In a preliminary study, WSSV binding protein (WBP) was specifically bound to the VP26 protein of WSSV. Therefore, we have developed the label-free affinity immunosensor using the WBP together with anti-GST-VP26 for quantitative detection of WSSV in shrimp pond water. When the biological molecules were immobilized on a gold electrode to form a self-assembled monolayer, it was then used to detect WSSV using a flow injection system with optimized conditions. Binding between the different copies of WSSV and the immobilized biological molecules was detected by an impedance change (ΔZ″) in real time. The sensitivity of the developed immunosensor was in the linear range of 1.6 × 10(1)-1.6 × 10(6) copies/μl. The system was highly sensitive for the analysis of WSSV as shown by the lack of impedance change when using yellow head virus (YHV). The developed immunosensor could be reused up to 37 times (relative standard deviation (RSD), 3.24 %) with a good reproducibility of residual activity (80-110 %). The immunosensor was simple to operate, reliable, reproducible, and could be applied for the detection and quantification of WSSV in water during shrimp cultivation.

A comparative study of x-ray emission from laser spots in laser-heated hohlraums relative to spots on simple disk targets

International Nuclear Information System (INIS)

Ze, F.; Langer, S.H.; Kauffman, R.L.; Kilkenny, J.D.; Landen, O.; Ress, D.; Rosen, M.D.; Suter, L.J.; Wallace, R.J.; Wiedwald, J.D.

1997-01-01

In this paper we report the results of experiments that compare the x-ray emission from a laser spot in a radiation-filled hohlraum to that from a similar laser spot on a simple disk target. The studies were done using the Nova laser facility [J. D. Lindl, Phys. Plasmas 2, 3933 (1995)] in its 0.35 μm wavelength, 1 ns square pulse configuration. Focal spot intensities were 2 endash 3.5x10 15 W/cm 2 . X-ray images measured x-ray conversion in a hohlraum and from an isolated disk simultaneously. A laser spot inside a hohlraum emitted more x rays, after subtracting the background emission from the hohlraum walls, than a spot on a disk. Numerical models suggest the enhanced spot emission inside the hohlraum is due to an increase in lateral transport relative to the disk. Filamentation in the hohlraum will also increase the spot size. The models agree fairly well with the results on spot spreading but do not explain the overall increase in conversion efficiency. copyright 1997 American Institute of Physics

Mercury-induced biochemical and proteomic changes in rice roots.

Science.gov (United States)

Chen, Yun-An; Chi, Wen-Chang; Huang, Tsai-Lien; Lin, Chung-Yi; Quynh Nguyeh, Thi Thuy; Hsiung, Yu-Chywan; Chia, Li-Chiao; Huang, Hao-Jen

2012-06-01

Mercury (Hg) is a serious environmental pollution threats to the planet. Accumulation of Hg in plants disrupts many cellular-level functions and inhibits growth and development, but the mechanism is not fully understood. We investigated cellular, biochemical and proteomic changes in rice roots under Hg stress. Root growth rate was decreased and Hg, reactive oxygen species (ROS), and malondialdehyde (MDA) content and lipoxygenase activity were increased significantly with increasing Hg concentration in roots. We revealed a time-dependent alteration in total glutathione content and enzymatic activity of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and peroxidase (POD) during Hg stress. 2-D electrophoresis revealed differential expression of 25 spots with Hg treatment of roots: 14 spots were upregulated and 11 spots downregulated. These differentially expressed proteins were identified by ESI-MS/MS to be involved in cellular functions including redox and hormone homeostasis, chaperone activity, metabolism, and transcription regulation. These results may provide new insights into the molecular basis of the Hg stress response in plants. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

Deciphering the Metabolic Changes Associated with Diapause Syndrome and Cold Acclimation in the Two-Spotted Spider Mite Tetranychus urticae

Science.gov (United States)

Khodayari, Samira; Moharramipour, Saeid; Larvor, Vanessa; Hidalgo, Kévin; Renault, David

2013-01-01

Diapause is a common feature in several arthropod species that are subject to unfavorable growing seasons. The range of environmental cues that trigger the onset and termination of diapause, in addition to associated hormonal, biochemical, and molecular changes, have been studied extensively in recent years; however, such information is only available for a few insect species. Diapause and cold hardening usually occur together in overwintering arthropods, and can be characterized by recording changes to the wealth of molecules present in the tissue, hemolymph, or whole body of organisms. Recent technological advances, such as high throughput screening and quantification of metabolites via chromatographic analyses, are able to identify such molecules. In the present work, we examined the survival ability of diapausing and non-diapausing females of the two-spotted spider mite, Tetranychus urticae, in the presence (0 or 5°C) or absence of cold acclimation. Furthermore, we examined the metabolic fingerprints of these specimens via gas chromatography-mass spectrophotometry (GC-MS). Partial Least Square Discriminant Analysis (PLS-DA) of metabolites revealed that major metabolic variations were related to diapause, indicating in a clear cut-off between diapausing and non-diapausing females, regardless of acclimation state. Signs of metabolic depression were evident in diapausing females, with most amino acids and TCA cycle intermediates being significantly reduced. Out of the 40 accurately quantified metabolites, seven metabolites remained elevated or were accumulated in diapausing mites, i.e. cadaverine, gluconolactone, glucose, inositol, maltose, mannitol and sorbitol. The capacity to accumulate winter polyols during cold-acclimation was restricted to diapausing females. We conclude that the induction of increased cold hardiness in this species is associated with the diapause syndrome, rather than being a direct effect of low temperature. Our results provide novel

PENGARUH KURS SPOT DAN KURS FORWARD DALAM MEMPREDIKSI FUTURE SPOT PADA PASAR VALAS KAWASAN ASIA TENGGARA

Directory of Open Access Journals (Sweden)

I Gusti Ayu Kade Diana Yanthi

2013-10-01

Full Text Available The importance of the activity in foreign exchange occur as the result of the development of international trading and increased international money and capital movement. The vast development of international economic  caused  economic relationship  among  countries  will  be  linked  to  each other  and  subsequently improve trading activities in their goods, money and capital flows. The variables used in this study are spot, forward  and future  spot. Based  on  the result  of Regression  Estimation  applied in  hypothesis on foreign currencies exchange which done simultaneously are proven able to predict future spot. The investor and international economic  actor could use spot  exchange rate and forward  exchange rate as a  short terms predictor for the next trimester in 2011.

Lipid-protein interaction induced domains: Kinetics and conformational changes in multicomponent vesicles

Science.gov (United States)

Sreeja, K. K.; Sunil Kumar, P. B.

2018-04-01

The spatio-temporal organization of proteins and the associated morphological changes in membranes are of importance in cell signaling. Several mechanisms that promote the aggregation of proteins at low cell surface concentrations have been investigated in the past. We show, using Monte Carlo simulations, that the affinity of proteins for specific lipids can hasten their aggregation kinetics. The lipid membrane is modeled as a dynamically triangulated surface with the proteins defined as in-plane fields at the vertices. We show that, even at low protein concentrations, strong lipid-protein interactions can result in large protein clusters indicating a route to lipid mediated signal amplification. At high protein concentrations, the domains form buds similar to that seen in lipid-lipid interaction induced phase separation. Protein interaction induced domain budding is suppressed when proteins act as anisotropic inclusions and exhibit nematic orientational order. The kinetics of protein clustering and resulting conformational changes are shown to be significantly different for the isotropic and anisotropic curvature inducing proteins.

Spreaders for immersion nucleate boiling cooling of a computer chip with a central hot spot

International Nuclear Information System (INIS)

Ali, Amir F.; El-Genk, Mohamed S.

2012-01-01

Highlights: ► The paper introduces a spreader concept for cooling high power chip with a hot spot. ► Spreader is comprised of a Cu substrate and copper micro-porous surface. ► Spreaders surface is cooled by nucleate boiling of PF-5060 dielectric liquid. ► Analysis demonstrated spreader effectiveness for mitigating hot spot effect. - Abstract: This paper numerically investigates the performance of composite spreaders comprised of Cu substrates and Cu micro-porous surfaces of different thicknesses for immersion cooling of 10 × 10 mm underlying computer chip with a 2 × 2 mm central hot spot. The local heat flux at the hot spot is three times the chip’s surface average outside the hot spot. The thickness of the Cu substrate changes from 1.6 to 3.2 mm and that of the Cu micro-porous surface changes from 80 to 197 μm. The spreaders are cooled by saturation nucleate boiling of PF-5060 dielectric liquid. The local values of the nucleate boiling heat transfer coefficients on the various Cu micro-porous surfaces are based on pool boiling experimental measurements. Results demonstrated the effectiveness of immersion cooling nucleate boiling for mitigating the effect of the hot spot. The spreaders decrease the maximum surface temperature and the temperature gradient on the chip surface and increase the dissipated thermal power by the chip and removed from the spreader surface. Increasing the thickness of the Cu substrate and/or decreasing the thickness of the Cu micro-porous surface increases the total thermal power removed, the chip surface temperature and the spreader’s footprint area.

Laser Spot Detection Based on Reaction Diffusion

Directory of Open Access Journals (Sweden)

Alejandro Vázquez-Otero

2016-03-01

Full Text Available Center-location of a laser spot is a problem of interest when the laser is used for processing and performing measurements. Measurement quality depends on correctly determining the location of the laser spot. Hence, improving and proposing algorithms for the correct location of the spots are fundamental issues in laser-based measurements. In this paper we introduce a Reaction Diffusion (RD system as the main computational framework for robustly finding laser spot centers. The method presented is compared with a conventional approach for locating laser spots, and the experimental results indicate that RD-based computation generates reliable and precise solutions. These results confirm the flexibility of the new computational paradigm based on RD systems for addressing problems that can be reduced to a set of geometric operations.

Fish proteins as targets of ferrous-catalyzed oxidation: identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry.

Science.gov (United States)

Pazos, Manuel; da Rocha, Angela Pereira; Roepstorff, Peter; Rogowska-Wrzesinska, Adelina

2011-07-27

Protein oxidation in fish meat is considered to affect negatively the muscle texture. An important source of free radicals taking part in this process is Fenton's reaction dependent on ferrous ions present in the tissue. The aim of this study was to investigate the susceptibility of cod muscle proteins in sarcoplasmic and myofibril fractions to in vitro metal-catalyzed oxidation and to point out protein candidates that might play a major role in the deterioration of fish quality. Extracted control proteins and proteins subjected to free radicals generated by Fe(II)/ascorbate mixture were labeled with fluorescein-5-thiosemicarbazide (FTSC) to tag carbonyl groups and separated by two-dimensional gel electrophoresis. Consecutive visualization of protein carbonyl levels by capturing the FTSC signal and total protein levels by capturing the SyproRuby staining signal allowed us to quantify the relative change in protein carbonyl levels corrected for changes in protein content. Proteins were identified using MALDI-TOF/TOF mass spectrometry and homology-based searches. The results show that freshly extracted cod muscle proteins exhibit a detectable carbonylation background and that the incubation with Fe(II)/ascorbate triggers a further oxidation of both sarcoplasmic and myofibril proteins. Different proteins exhibited various degrees of sensitivity to oxidation processes. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), nucleoside diphosphate kinase B (NDK), triosephosphate isomerase, phosphoglycerate mutase, lactate dehydrogenase, creatine kinase, and enolase were the sarcoplasmic proteins most vulnerable to ferrous-catalyzed oxidation. Moreover, NDK, phosphoglycerate mutase, and GAPDH were identified in several spots differing by their pI, and those forms showed different susceptibilities to metal-catalyzed oxidation, indicating that post-translational modifications may change the resistance of proteins to oxidative damage. The Fe(II)/ascorbate treatment significantly

Rocky Mountain spotted fever, Colombia.

Science.gov (United States)

Hidalgo, Marylin; Orejuela, Leonora; Fuya, Patricia; Carrillo, Pilar; Hernandez, Jorge; Parra, Edgar; Keng, Colette; Small, Melissa; Olano, Juan P; Bouyer, Donald; Castaneda, Elizabeth; Walker, David; Valbuena, Gustavo

2007-07-01

We investigated 2 fatal cases of Rocky Mountain spotted fever that occurred in 2003 and 2004 near the same locality in Colombia where the disease was first reported in the 1930s. A retrospective serosurvey of febrile patients showed that > 21% of the serum samples had antibodies aaainst spotted fever group rickettsiae.

Age Spots

Science.gov (United States)

... for Every Season How to Choose the Best Skin Care Products In This Section Dermatologic Surgery What is dermatologic ... for Every Season How to Choose the Best Skin Care Products Age Spots Treatment Options Learn more about treatment ...

An improved microculture-hemolytic spot assay for the study of carrier-specific antibody responses.

Science.gov (United States)

Kotkes, P; Weisman, Z; Mozes, E; Bentwich, Z

1984-11-30

A microculture system based on limiting dilution and a hemolytic spot assay was adapted for study of the carrier-specific anti-hapten response in vitro. Spleen or lymph node cells from normal mice or mice immunized with NIP-ovalbumin (NIP-OVA) or NIP-human thyroglobulin (NIP-Tg) were cultured for 5 days by the microculture technique. The anti-hapten (anti-NIP) response was measured by assaying the supernatants of the microcultures in a hemolytic spot test with NIP coupled to sheep red blood cells. A micro-ELISA reader was adapted to read the degree of lysis in the spot assay which gives an objective quantitation of the degree of lysis and thus reduces the number of culture replicates. In vivo induced specific helper cells in mice immunized with the carrier protein, human thyroglobulin, as well as carrier-specific T cell factors, gave rise to carrier-specific anti-NIP responses. The microculture system may enhance the expression of T-cell helper function when suppressor cells or their precursors are present in the initial cell preparation.

Anisotropic multi-spot DBR porous silicon chip for the detection of human immunoglobin G.

Science.gov (United States)

Cho, Bomin; Um, Sungyong; Sohn, Honglae

2014-07-01

Asymmetric porous silicon multilayer (APSM)-based optical biosensor was developed to specify human Immunoglobin G (Ig G). APSM chip was generated by an electrochemical etching of silicon wafer using an asymmetric electrode configuration in aqueous ethanolic HF solution and constituted with nine arrayed porous silicon multilayer. APSM prepared from anisotropic etching conditions displayed a sharp reflection resonance in the reflectivity spectrum. Each spot displayed single reflection resonance at different wavelengths as a function of the lateral distance from the Pt counter electrode. The sensor system was consisted of the 3 x 3 spot array of APSM modified with protein A. The system was probed with an aqueous human Ig G. Molecular binding and specificity was monitored as a shift in wavelength of reflection resonance.

Splitting of turbulent spot in transitional pipe flow

Science.gov (United States)

Wu, Xiaohua; Moin, Parviz; Adrian, Ronald J.

2017-11-01

Recent study (Wu et al., PNAS, 1509451112, 2015) demonstrated the feasibility and accuracy of direct computation of the Osborne Reynolds' pipe transition problem without the unphysical, axially periodic boundary condition. Here we use this approach to study the splitting of turbulent spot in transitional pipe flow, a feature first discovered by E.R. Lindgren (Arkiv Fysik 15, 1959). It has been widely believed that spot splitting is a mysterious stochastic process that has general implications on the lifetime and sustainability of wall turbulence. We address the following two questions: (1) What is the dynamics of turbulent spot splitting in pipe transition? Specifically, we look into any possible connection between the instantaneous strain rate field and the spot splitting. (2) How does the passive scalar field behave during the process of pipe spot splitting. In this study, the turbulent spot is introduced at the inlet plane through a sixty degree wide numerical wedge within which fully-developed turbulent profiles are assigned over a short time interval; and the simulation Reynolds numbers are 2400 for a 500 radii long pipe, and 2300 for a 1000 radii long pipe, respectively. Numerical dye is tagged on the imposed turbulent spot at the inlet. Splitting of the imposed turbulent spot is detected very easily. Preliminary analysis of the DNS results seems to suggest that turbulent spot slitting can be easily understood based on instantaneous strain rate field, and such spot splitting may not be relevant in external flows such as the flat-plate boundary layer.

Effects of Nickel, Chlorpyrifos and Their Mixture on the Dictyostelium discoideum Proteome

Science.gov (United States)

Boatti, Lara; Robotti, Elisa; Marengo, Emilio; Viarengo, Aldo; Marsano, Francesco

2012-01-01

Mixtures of chemicals can have additive, synergistic or antagonistic interactions. We investigated the effects of the exposure to nickel, the organophosphate insecticide chlorpyrifos at effect concentrations (EC) of 25% and 50% and their binary mixture (Ec25 + EC25) on Dictyostelium discoideum amoebae based on lysosomal membrane stability (LMS). We treated D. discoideum with these compounds under controlled laboratory conditions and evaluated the changes in protein levels using a two-dimensional gel electrophoresis (2DE) proteomic approach. Nickel treatment at EC25 induced changes in 14 protein spots, 12 of which were down-regulated. Treatment with nickel at EC50 resulted in changes in 15 spots, 10 of which were down-regulated. Treatment with chlorpyrifos at EC25 induced changes in six spots, all of which were down-regulated; treatment with chlorpyrifos at EC50 induced changes in 13 spots, five of which were down-regulated. The mixture corresponding to EC25 of each compound induced changes in 19 spots, 13 of which were down-regulated. The data together reveal that a different protein expression signature exists for each treatment, and that only a few proteins are modulated in multiple different treatments. For a simple binary mixture, the proteomic response does not allow for the identification of each toxicant. The protein spots that showed significant differences were identified by mass spectrometry, which revealed modulations of proteins involved in metal detoxification, stress adaptation, the oxidative stress response and other cellular processes. PMID:23443088

Enhanced processing of SPOT multispectral satellite imagery for environmental monitoring and modelling

Energy Technology Data Exchange (ETDEWEB)

Clark, B.

2010-07-01

The Taita Hills in southeastern Kenya form the northernmost part of Africa's Eastern Arc Mountains, which have been identified by Conservation International as one of the top ten biodiversity hotspots on Earth. As with many areas of the developing world, over recent decades the Taita Hills have experienced significant population growth leading to associated major changes in land use and land cover (LULC), as well as escalating land degradation, particularly soil erosion. Multi-temporal medium resolution multispectral optical satellite data, such as imagery from the SPOT HRV, HRVIR, and HRG sensors, provides a valuable source of information for environmental monitoring and modelling at a landscape level at local and regional scales. However, utilization of multi-temporal SPOT data in quantitative remote sensing studies requires the removal of atmospheric effects and the derivation of surface reflectance factor (rho{sub s}). Furthermore, for areas of rugged terrain, such as the Taita Hills, topographic correction is necessary to derive comparable (rho{sub s}) throughout a SPOT scene. Reliable monitoring of LULC change over time and modelling of land degradation and human population distribution and abundance are of crucial importance to sustainable development, natural resource management, biodiversity conservation, and understanding and mitigating climate change and its impacts. The main purpose of this thesis was to develop and validate enhanced processing of SPOT satellite imagery for use in environmental monitoring and modelling at a landscape level, in regions of the developing world with limited ancillary data availability. The Taita Hills formed the application study site, whilst the Helsinki metropolitan region was used as a control site for validation and assessment of the applied atmospheric correction techniques, where multiangular (rho{sub s}) field measurements were taken and where horizontal visibility meteorological data concurrent with image

Increase in local protein concentration by field-inversion gel electrophoresis

Directory of Open Access Journals (Sweden)

Paulus Aran

2007-09-01

Full Text Available Abstract Background Proteins that migrate through cross-linked polyacrylamide gels (PAGs under the influence of a constant electric field experience negative factors, such as diffusion and non-specific trapping in the gel matrix. These negative factors reduce protein concentrations within a defined gel volume with increasing migration distance and, therefore, decrease protein separation efficiency. Enhancement of protein separation efficiency was investigated by implementing pulsed field-inversion gel electrophoresis (FIGE. Results Separation of model protein species and large protein complexes was compared between FIGE and constant field electrophoresis (CFE in different percentages of PAGs. Band intensities of proteins in FIGE with appropriate ratios of forward and backward pulse times were superior to CFE despite longer running times. These results revealed an increase in band intensity per defined gel volume. A biphasic protein relative mobility shift was observed in percentages of PAGs up to 14%. However, the effect of FIGE on protein separation was stochastic at higher PAG percentage. Rat liver lysates subjected to FIGE in the second-dimension separation of two-dimensional polyarcylamide gel electrophoresis (2D PAGE showed a 20% increase in the number of discernible spots compared with CFE. Nine common spots from both FIGE and CFE were selected for peptide sequencing by mass spectrometry (MS, which revealed higher final ion scores of all nine protein spots from FIGE. Native protein complexes ranging from 800 kDa to larger than 2000 kDa became apparent using FIGE compared with CFE. Conclusion The present investigation suggests that FIGE under appropriate conditions improves protein separation efficiency during PAGE as a result of increased local protein concentration. FIGE can be implemented with minimal additional instrumentation in any laboratory setting. Despite the tradeoff of longer running times, FIGE can be a powerful protein

Increase in local protein concentration by field-inversion gel electrophoresis.

Science.gov (United States)

Tsai, Henghang; Low, Teck Yew; Freeby, Steve; Paulus, Aran; Ramnarayanan, Kalpana; Cheng, Chung-Pui Paul; Leung, Hon-Chiu Eastwood

2007-09-26

Proteins that migrate through cross-linked polyacrylamide gels (PAGs) under the influence of a constant electric field experience negative factors, such as diffusion and non-specific trapping in the gel matrix. These negative factors reduce protein concentrations within a defined gel volume with increasing migration distance and, therefore, decrease protein separation efficiency. Enhancement of protein separation efficiency was investigated by implementing pulsed field-inversion gel electrophoresis (FIGE). Separation of model protein species and large protein complexes was compared between FIGE and constant field electrophoresis (CFE) in different percentages of PAGs. Band intensities of proteins in FIGE with appropriate ratios of forward and backward pulse times were superior to CFE despite longer running times. These results revealed an increase in band intensity per defined gel volume. A biphasic protein relative mobility shift was observed in percentages of PAGs up to 14%. However, the effect of FIGE on protein separation was stochastic at higher PAG percentage. Rat liver lysates subjected to FIGE in the second-dimension separation of two-dimensional polyarcylamide gel electrophoresis (2D PAGE) showed a 20% increase in the number of discernible spots compared with CFE. Nine common spots from both FIGE and CFE were selected for peptide sequencing by mass spectrometry (MS), which revealed higher final ion scores of all nine protein spots from FIGE. Native protein complexes ranging from 800 kDa to larger than 2000 kDa became apparent using FIGE compared with CFE. The present investigation suggests that FIGE under appropriate conditions improves protein separation efficiency during PAGE as a result of increased local protein concentration. FIGE can be implemented with minimal additional instrumentation in any laboratory setting. Despite the tradeoff of longer running times, FIGE can be a powerful protein separation tool.

Proteomic Identification of Target Proteins of Thiodigalactoside in White Adipose Tissue from Diet-Induced Obese Rats

Directory of Open Access Journals (Sweden)

Hilal Ahmad Parray

2015-06-01

Full Text Available Previously, galectin-1 (GAL1 was found to be up-regulated in obesity-prone subjects, suggesting that use of a GAL1 inhibitor could be a novel therapeutic approach for treatment of obesity. We evaluated thiodigalactoside (TDG as a potent inhibitor of GAL1 and identified target proteins of TDG by performing comparative proteome analysis of white adipose tissue (WAT from control and TDG-treated rats fed a high fat diet (HFD using two dimensional gel electrophoresis (2-DE combined with MALDI-TOF-MS. Thirty-two spots from a total of 356 matched spots showed differential expression between control and TDG-treated rats, as identified by peptide mass fingerprinting. These proteins were categorized into groups such as carbohydrate metabolism, tricarboxylic acid (TCA cycle, signal transduction, cytoskeletal, and mitochondrial proteins based on functional analysis using Protein Annotation Through Evolutionary Relationship (PANTHER and Database for Annotation, Visualization, Integrated Discovery (DAVID classification. One of the most striking findings of this study was significant changes in Carbonic anhydrase 3 (CA3, Voltage-dependent anion channel 1 (VDAC1, phosphatidylethanolamine-binding protein 1 (PEBP1, annexin A2 (ANXA2 and lactate dehydrogenase A chain (LDHA protein levels between WAT from control and TDG-treated groups. In addition, we confirmed increased expression of thermogenic proteins as well as reduced expression of lipogenic proteins in response to TDG treatment. These results suggest that TDG may effectively prevent obesity, and TDG-responsive proteins can be used as novel target proteins for obesity treatment.

Stability of benzodiazepines and cocaine in blood spots stored on filter paper.

Science.gov (United States)

Alfazil, Abdulkareem A; Anderson, Robert A

2008-09-01

Previous studies have shown that drug concentrations in blood can change during storage, especially at room temperature, but even labile drugs such as cocaine may be stable in dried blood spots (DBS). A new method has been developed for the analysis of hydrolytically labile drugs in blood spots on filter paper in order to assess their degradation during a storage period of one month. The drugs selected included flunitrazepam, temazepam, oxazepam, lorazepam, nitrazepam, diazepam, and cocaine. A Guthrie card 903 was spotted with 100 microL of blood containing the drugs at concentrations of 1000 ng/mL and left overnight to dry at room temperature. The filter paper was suspended in extraction buffer for 1 h with ultrasonication. Drugs were then extracted from the buffer by solid-phase extraction using Clean Screen((R)) columns and analyzed by liquid chromatography-tandem mass spectrometry. Method validation showed that all calibration curves were linear over the concentration range 5-200 ng/spot with correlation coefficients of 0.994-0.999. Interday and intraday precisions at three concentrations (10, 50, and 100 ng/spot) were 1.6-18.3% and 2.8-14.7%, respectively. Limits of detection were 0.29-0.74 ng/spot, and lower limits of quantitation were 0.99-2.46 ng/spot. Recoveries of all analytes were in the range 81-106%. DBS were stored in duplicate at room temperature, 4 degrees C, and -20 degrees C for up to one month. Degradation of the drugs in DBS at all storage conditions was less than for the corresponding liquid blood samples stored under similar conditions and more than 80% of each analyte could be recovered from the samples.

Laser based spot weld characterization

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Jonietz, Florian; Myrach, Philipp; Rethmeier, Michael; Suwala, Hubert; Ziegler, Mathias

2016-02-01

Spot welding is one of the most important joining technologies, especially in the automotive industry. Hitherto, the quality of spot welded joints is tested mainly by random destructive tests. A nondestructive testing technique offers the benefit of cost reduction of the testing procedure and optimization of the fabrication process, because every joint could be examined. This would lead to a reduced number of spot welded joints, as redundancies could be avoided. In the procedure described here, the spot welded joint between two zinc-coated steel sheets (HX340LAD+Z100MB or HC340LA+ZE 50/50) is heated optically on one side. Laser radiation and flash light are used as heat sources. The melted zone, the so called "weld nugget" provides the mechanical stability of the connection, but also constitutes a thermal bridge between the sheets. Due to the better thermal contact, the spot welded joint reveals a thermal behavior different from the surrounding material, where the heat transfer between the two sheets is much lower. The difference in the transient thermal behavior is measured with time resolved thermography. Hence, the size of the thermal contact between the two sheets is determined, which is directly correlated to the size of the weld nugget, indicating the quality of the spot weld. The method performs well in transmission with laser radiation and flash light. With laser radiation, it works even in reflection geometry, thus offering the possibility of testing with just one-sided accessibility. By using heating with collimated laser radiation, not only contact-free, but also remote testing is feasible. A further convenience compared to similar thermographic approaches is the applicability on bare steel sheets without any optical coating for emissivity correction. For this purpose, a proper way of emissivity correction was established.

Advanced spot quality analysis in two-colour microarray experiments

Directory of Open Access Journals (Sweden)

Vetter Guillaume

2008-09-01

Full Text Available Abstract Background Image analysis of microarrays and, in particular, spot quantification and spot quality control, is one of the most important steps in statistical analysis of microarray data. Recent methods of spot quality control are still in early age of development, often leading to underestimation of true positive microarray features and, consequently, to loss of important biological information. Therefore, improving and standardizing the statistical approaches of spot quality control are essential to facilitate the overall analysis of microarray data and subsequent extraction of biological information. Findings We evaluated the performance of two image analysis packages MAIA and GenePix (GP using two complementary experimental approaches with a focus on the statistical analysis of spot quality factors. First, we developed control microarrays with a priori known fluorescence ratios to verify the accuracy and precision of the ratio estimation of signal intensities. Next, we developed advanced semi-automatic protocols of spot quality evaluation in MAIA and GP and compared their performance with available facilities of spot quantitative filtering in GP. We evaluated these algorithms for standardised spot quality analysis in a whole-genome microarray experiment assessing well-characterised transcriptional modifications induced by the transcription regulator SNAI1. Using a set of RT-PCR or qRT-PCR validated microarray data, we found that the semi-automatic protocol of spot quality control we developed with MAIA allowed recovering approximately 13% more spots and 38% more differentially expressed genes (at FDR = 5% than GP with default spot filtering conditions. Conclusion Careful control of spot quality characteristics with advanced spot quality evaluation can significantly increase the amount of confident and accurate data resulting in more meaningful biological conclusions.

Proteomics analysis of ram sperm by heavy ion radiation

International Nuclear Information System (INIS)

He Yuxuan; Li Hongyan; Zhang Hong

2013-01-01

The objective of this study was to investigate the proteome changes induced by heavy ion radiation using irradiated ram sperm by a two-dimensional electrophoresis (2-DE) analysis. The 2D gels were stained with Coomassie Brilliant Blue. Differentially expressed proteins were detected by PDQuest 8.0 software and subjected to ion trap mass spectrometer equipped with a surveyor HPLC system, and differential protein spots were identified. Results showed there are five differential protein spots in irradiated sperm gels, four up-regulated protein spots and one spot missed. The differentially expressed protein spots were identified to be two up-regulated proteins including enolase, and enolase 1. It was concluded there was proteome changes induced by heavy ion radiation in ram sperm, which may be useful to clarify the physiology state of ram sperm in heavy ion radiation and provide a theoretical basis for radiation ram breeding. (authors)

Detecting hot spots at hazardous-waste sites

International Nuclear Information System (INIS)

Zirschky, J.; Gilbert, R.O.

1984-01-01

Evaluating the need for remedial cleanup at a waste site involves both finding the average contaminant concentration and identifying highly contaminated areas, or hot spots. A nomographic procedure to determine the sample configuration needed to locate a hot spot is presented. The technique can be used to develop a waste-site sampling plant - to determine either the grid spacing required to detect a hot spot at a given level of confidence, or the probability of finding a hot spot of a certain size, given a particular grid spacing. The method and computer program (ELIPGRID) were developed for locating geologic deposits, but the basic procedure can also be used to detect hot spots at chemical- or nuclear-waste disposal sites. Nomographs based on the original program are presented for three sampling-grid configurations - square, rectangular and triangular

Perceptual changes and drivers of liking in high protein extruded snacks.

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Kreger, Joseph W; Lee, Youngsoo; Lee, Soo-Yeun

2012-04-01

Increasing the amount of protein in snack foods can add to their satiating ability, which aligns with many health-based trends currently seen in the food industry. Understanding the effect of adding high levels of protein in a food matrix is essential for product development. The objective for this research was to determine the effects of varying protein type and level on the sensory-related aspects of a model extruded snack food. Independent variables in the design of the snacks were the level of total protein and the protein type in the formulation. The level of protein ranged from 28% to 43% (w/w) in 5% increments. The protein type varied in the ratio of whey to soy protein ranging from 0: 100 to 100: 0, in 25% increments. Descriptive analysis was conducted on the samples to profile their sensory characteristics. Protein type was found to be the predominant variable in differentiating the sensory characteristics of the samples. Soy protein imparted nutty, grainy aromas-by-mouth, and increased expansion during processing, resulting in a lighter, crispier texture. Whey protein imparted dairy related aromas-by-mouth and inhibited expansion during processing, resulting in a more dense, crunchy texture. Separately, 100 consumers rated their acceptance of the samples using the 9-point hedonic scale. It was found that protein type was also the predominant variable in affecting acceptance, with some clusters of consumers preferring samples comprised of soy protein, and others preferring samples with whey. Food product developers can use these findings to predict changes in a similar food product by varying protein level or protein type. This work shows how the perceivable appearance, aroma, and texture characteristics of puffed snack foods change when adding protein or changing the protein type. The type of protein incorporated was shown to have major effects on the characteristics of the snacks, partially because of their impact on how much the snacks puffed during

Sequence charge decoration dictates coil-globule transition in intrinsically disordered proteins

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