bluuebunny/crossref_metadata_2025 huggingface dataset:
Inactivated Orf virus (Parapoxvirus ovis) elicits antifibrotic activity in models of liver fibrosis
Authors: Nowatzky Janina , Knorr Andreas , Hirth‐Dietrich Claudia , Siegling Angela , Volk Hans‐Dieter , Limmer Andreas , Knolle Percy , Weber Olaf
Published: September 2012

Aim

Inactivated Orf virus (ORFV, Parapoxvirus ovis) demonstrates strong antiviral activity in animal models including a human hepatitis B virus (HBV)‐transgenic mouse. In addition, expression of interferon (IFN)‐γ and interleukin‐10 (IL‐10) was induced after administration of inactivated ORFV in these mice. IFN‐γ and IL‐10 are known to elicit antifibrotic activity. We therefore aimed to study antifibrotic activity of inactivated ORFV in models of liver fibrosis.

Methods

We characterized ORFV‐induced hepatic cytokine expression in rats. We then studied ORFV in two models of liver fibrosis in rats, pig serum‐induced liver fibrosis and carbon tetrachloride (CCL4)‐induced liver fibrosis.

Results

ORFV induced hepatic expression of IFN‐γ and IL‐10 in rats. ORFV mediated antifibrotic activity when administrated concomitantly with the fibrosis‐inducing agents in both models of liver fibrosis. Importantly, when CCL4‐induced liver fibrosis was already established, ORFV application still showed significant antifibrotic activity. In addition, we were able to demonstrate a direct antifibrotic effect of ORFV on stellate cells.

Conclusion

These results establish a potential novel antifibrotic therapeutic approach that not only prevents but also resolves established liver fibrosis. Further studies are required to unravel the details of the mechanisms involved.
Role of gut microbiota in liver diseases
Authors: Miyake Yasuhiro , Yamamoto Kazuhide
Published: September 2012

The liver constantly encounters food‐derived antigens and bacterial components such as lipopolysaccharide translocated from the gut into the portal vein. Bacterial components stimulate Toll‐like receptors (TLR), which are expressed on Kupffer cells, biliary epithelial cells, hepatocytes, hepatic stellate cells, endothelial cells and dendritic cells and recognize specific pathogen‐associated molecular patterns. The signaling of TLR to its main ligand triggers inflammation. Usually, in order to protect against hyperactivation of the immune system and to prevent organ failure by persistent inflammation, TLR tolerance to repeated stimuli is induced. In chronic liver diseases, a breakdown in TLR tolerance occurs. Furthermore, Kupffer cells, hepatic stellate cells and natural killer T cells are key components of innate immunity. Decreased numbers and impaired ability of these cells lead to failures in immune tolerance, resulting in persistent inflammation. Recently, the activation of inflammasome was revealed to control the secretion of pro‐inflammatory cytokines such as interleukin‐1β in response to bacterial pathogens. Innate immunity seems to be an important contributor to the pathogenesis of fatty liver disease and autoimmune liver disease. Recently, probiotics were reported to affect various liver diseases via shifts in gut microbiota and the stability of intestinal permeability. However, many unresolved questions remain. Further analysis will be needed to gain a more comprehensive understanding of the association of innate immunity with the pathogenesis of various liver diseases.
What Do FAS 157 “Fair Values” Really Measure: Value Or Risk?
Authors: Ronen Joshua
Published: September 2012

FAS 157, the U.S. accounting standard that prescribes how fair values of assets and liabilities are to be measured when other U.S. GAAP standards require fair valuation, stipulates that fair values be measured as the exit values of assets and liabilities—the proceeds for assets hypothetically sold on the date of the financial report, and, correspondingly, the amount required to settle liabilities on the date of the financial report. This conceptual article argues that exit values do not reflect the value of the net assets of the firm to shareholders, which is best reflected by discounted cash flows to maturity. Moreover, exit values—biasing fair values downward when markets are illiquid—have a pernicious, systemic risk effect; specifically, they give rise to write‐downs that in turn cause contagion: prices of equities and other financial instruments of peers react negatively, leading to further write‐downs by those peers. This may have aggravated the recent financial crisis. However, while exit values are not proper measures of value to shareholders, they are useful measures of downside risk when prospects turn sour for a firm. Thus, both exit values and discounted cash flows should be presented in financial statements.
Timing of Sale, Pricing, and Cost Information: Evidence from the Airline Industry*
Authors: Hsu Sylvia H. , Lee Johnny Jiungyee
Published: September 2012

This study examines the association between when an airline sells its passenger seats and the pricing method (marginal cost or full cost) it employs. Prior literature suggests that when firms are able to change prices during the selling period, the optimality of full cost pricing or marginal cost pricing depends on when demand information is revealed during the period between capacity commitment decisions and time of sale. Full cost‐based pricing is appropriate in determining capacity commitment and prices simultaneously, while marginal cost provides more relevant information for pricing when capacity has been committed. Using the price and cost data from a sample of four U.S. domestic airlines, we find that full cost explains price variations of first‐day sales robustly. The adjusted R2 of the marginal cost pricing model is larger in the sample of sales two days prior to departure than in the sample of first‐day sales. In the analysis of the sample of sales two days prior to departure, we find that, based on the adjusted R2 of the full cost pricing and marginal cost pricing models, the explanatory power of marginal cost pricing is relatively weaker than full cost pricing. Our results document the use of different cost information along the dynamic change of price and provide implications in understanding the role of cost information in setting prices.
Two Reasons Why Epistemic Reasons Are Not Object‐Given Reasons
Authors: BOOTH ANTHONY ROBERT
Published: September 2012

In this paper I discuss two claims; the first is the claim that state‐given reasons for belief are of a radically different kind to object‐given reasons for belief. The second is that, where this last claim is true, epistemic reasons are object‐given reasons for belief (EOG). I argue that EOG has two implausible consequences: (i) that suspension of judgement can never be epistemically justified, and (ii) that the reason that epistemically justifies a belief that p can never be the reason for which one believes that p.
Accounting for missing data when assessing availability in animal population surveys: an application to ice‐associated seals in theBeringSea
Authors: Conn Paul B. , Johnson Devin S. , London Josh M. , Boveng Peter L.
Published: September 2012

Population ecologists often use telemetry to estimate the probability that an animal is available for sampling (availability probability). Such estimates are paramount for generating reliable estimates of animal abundance and are sometimes of biological interest in their own right (e.g. for characterizing pinniped haul‐out behaviour or landbird singing frequency).

We consider the estimation of availability probability from telemetry data when there are missing records. When records are missing‐completely‐at‐random (i.e. not influenced by whether an animal is available for sampling or other covariates), approaches that censor missing records result in unbiased estimates of availability probability. However, censoring such records can result in bias when data are not missing‐completely‐at‐random.

We present a novelBayesian temporal availability model that can be used to explicitly account for the process by which missing data arise. Our approach couples an underlying model for the partially observed availability process together with an observation model that allows state‐dependent probabilities of missingness. This approach provides relatively unbiased estimates of availability probability even in the face of violations of the missing‐at‐random assumption. A new R package,TempOcc, is introduced to automate estimation.

We demonstrate the utility of our approach by analysing both simulated data and hourly satellite telemetry records for 157 ice‐associated seals in theBeringSea, for which 27% of records were missing. In this case, we were interested in generating unbiased estimates of haul‐out probabilities. Such probabilities are necessary when estimating abundance from aerial survey counts and assessing possible changes in seal phenology.

Our analysis indicates that the missing‐at‐random assumption is indeed violated in telemetry studies of phocid seals in theBeringSea. However, estimates of availability probability do not depend critically on this assumption.

We recommend that ecologists routinely consider the implications of violating the missing‐at‐random assumption when estimating availability probabilities. Where possible, the modelling framework articulated in this study can be used to diagnose and correct for violations of the missing‐at‐random assumption.
Active halogen compounds and proteinaceous material: loss of activity of topical anti-infectives by halogen consumption
Authors: Gottardi Waldemar , Nagl Markus
Published: September 2012

Objectives

The activity of oxidants, such as halogens and active halogen compounds, decreases generally in the presence of proteinaceous material. A quantification of consumption effects was performed to judge the suitability of different representatives as antiseptics and their compatibility with pharmaceutical additives.

Methods

An iodometric approach served to assess the temporal loss of oxidation capacity in the presence of peptone and fetal calf serum (FCS). The tested agents comprised active halogen compounds, well-known and in particular novel chloramine-based agents indicated for the topical treatment of infections.

Key findings

The decrease in oxidation capacity was higher in the presence of FCS than of peptone and correlated with the reactivity of the oxidants in both cases. The highest consumption rates were for active bromine compounds followed by hypochlorous acid and heterocyclic chlorimides, such as dichloro-isocyanuric acid, while N-chlorotaurine and related amine-based analogues were least consumed. The pH dependence was only remarkable for chloramine T.

Conclusions

The observed consumption effects are the result of the differing oxidizing (chlorinating) potencies. Since consumption and irritation are founded on the very same reactions, representatives with low reactivity (N-chloro amino acids) are regarded as more tolerable and retain more oxidative capacity, which provides a more sustained antimicrobial activity.
An assessment of linkage disequilibrium in Holstein cattle using a Bayesian network
Authors: Morota G. , Valente B.D. , Rosa G.J.M. , Weigel K.A. , Gianola D.
Published: September 2012

Linkage disequilibrium (LD) is defined as a non‐random association of the distributions of alleles at different loci within a population. This association between loci is valuable in prediction of quantitative traits in animals and plants and in genome‐wide association studies. A question that arises is whether standard metrics such as D′ and r2 reflect complex associations in a genetic system properly. It seems reasonable to take the view that loci associate and interact together as a system or network, as opposed to in a simple pairwise manner. We used a Bayesian network (BN) as a representation of choice for an LD network. A BN is a graphical depiction of a probability distribution and can represent sets of conditional independencies. Moreover, it provides a visual display of the joint distribution of the set of random variables in question. The usefulness of BN for linkage disequilibrium was explored and illustrated using genetic marker loci found to have the strongest effects on milk protein in Holstein cattle based on three strategies for ranking marker effect estimates: posterior means, standardized posterior means and additive genetic variance. Two different algorithms, Tabu search (a local score–based algorithm) and incremental association Markov blanket (a constraint‐based algorithm), coupled with the chi‐square test, were used for learning the structure of the BN and were compared with the reference r2 metric represented as an LD heat map. The BN captured several genetic markers associated as clusters, implying that markers are inter‐related in a complicated manner. Further, the BN detected conditionally dependent markers. The results confirm that LD relationships are of a multivariate nature and that r2 gives an incomplete description and understanding of LD. Use of an LD Bayesian network enables inferring associations between loci in a systems framework and provides a more accurate picture of LD than that resulting from the use of pairwise metrics.
L1CAM and its cell‐surface mutants: new mechanisms and effects relevant to the physiology and pathology of neural cells
Authors: Tagliavacca Luigina , Colombo Federico , Racchetti Gabriella , Meldolesi Jacopo
Published: September 2012

The L1 syndrome, a genetic disease that affects 1/30 000 newborn males, is sustained by numerous missense mutations of L1 cell adhesion molecule (L1CAM), an adhesion surface protein active also in transmembrane signaling, essential for the development and function of neurons. To investigate the cell biology of L1CAM, we employed a high RE1‐silencing transcription (factor) clone of the pheochromocytoma PC12 line, defective in L1CAM expression and neurite outgrowth. The clone was transfected with wild‐type L1CAM and four missense, disease‐inducing point mutants encoding proteins distributed to the cell surface. The mutant‐expressing cells, defective in adhesion to extracellular matrix proteins and in migration, exhibited unchanged proliferation. The nerve growth factor (NGF)‐induced neurite outgrowth was re‐established in defective clone cells transfected with the wild‐type and the H210Q and I219T L1CAMs mutants, but not in the others. The stimulated outgrowth was confirmed in a second defective PC12 clone over‐expressing the NGF receptor TrkA, treated with NGF and/or a recombinant L1CAM chimera. These results revealed a new function of L1CAM, a positive, robust and dose‐dependent modulation of the TrkA receptor activated spontaneously or by NGF. The variable effects observed with the different L1CAM mutants suggest that this function contributes to the marked heterogeneity of symptoms and severity observed in the patients affected by the L1 syndrome.
Pen‐2 is dispensable for endoproteolysis of presenilin 1, and nicastrin‐Aph subcomplex is important for both γ‐secretase assembly and substrate recruitment
Authors: Mao Guozhang , Cui Mei‐Zhen , Li Tong , Jin Yipeng , Xu Xuemin
Published: September 2012

γ‐secretase is a protease complex with at least four components: presenilin, nicastrin (NCT), anterior pharynx‐defective 1 (Aph‐1), and presenilin enhancer 2 (Pen‐2). In this study, using knockout cell lines and small interfering RNA technology, our data demonstrated that the disappeared presenilin 1 C‐terminal fragment (PS1C) caused by knockdown of pen‐2 or knockout of NCT or Aph‐1 was recovered by the addition of proteasome inhibitors, indicating that Pen‐2, as well as NCT and Aph‐1α, is dispensable for presenilin endoproteolysis. Our data also demonstrate that the formation of the nicastrin‐Aph‐1 subcomplex plays not only an important role in γ‐secretase complex assembly but also in recruiting substrate C‐terminal fragment of amyloid precursor protein generated by β‐cleavage. Ablating any one component resulted in the instability of other components of the γ‐secretase complex, and the presence of all three of the other components is required for full maturation of NCT.
Impact of Grapes Heterogeneity According to Sugar Level on both Physical and Mechanical Berries Properties and Their Anthocyanins Extractability at Harvest
Authors: Zouid I. , Siret R. , Jourjon F. , Mehinagic E. , Rolle L.
Published: September 2012

Cabernet Franc grapes belonging to six different classes of sugar level content (from 176.5 to 258.5 g/L) at harvest were studied. Their physical proprieties and mechanical behavior were measured using compression and puncture tests. The evolution of anthocyanins extractability was determined. Significant differences were found in grape textural properties and anthocyanins content due to the sugar level content in the different samples of berries tested. It appears that the sweetest berries are characterized by a higher extraction yield of anthocyanins (up to +3.55%) with respect to those with more low sugar content. Furthermore, significant relationships exist between the extractability of anthocyanins and the mechanical and physical attributes of the grape berries. The best correlations were obtained with physical parameters such as the berry weight and the surface : volume ratio (r = −0.99 and 0.97, respectively) and with double compression test parameters such as hardness, chewiness and springiness (r = −0.90, −0.91 and 0.92, respectively).

Practical Applications

The purpose and the originality of this work were to determine the real impact of the heterogeneity of grapes at harvest. Among the berries with different sugar contents, were assessed the real differences in anthocyanin contents and extractability at harvest. The results acquired, also in function of different distribution percentage of the berries after floatation, suggest that each class of berries may influence in unlike way the final quality of wine. Therefore, automatic densimetric berry‐sorting machines recently developed and proposed for use in wineries could be indeed useful to select the grapes after the harvest.
ArchaealJAB1/MPN/MOV34 metalloenzyme (HvJAMM1) cleaves ubiquitin‐like small archaeal modifier proteins (SAMPs) from protein‐conjugates
Authors: Hepowit Nathaniel L. , Uthandi Sivakumar , Miranda Hugo V. , Toniutti Micaela , Prunetti Laurence , Olivarez Oliver , De Vera Ian Mitchelle S. , Fanucci Gail E. , Chen Sixue , Maupin‐Furlow Julie A.
Published: September 2012

Proteins withJAB1/MPN/MOV34 metalloenzyme (JAMM/MPN+) domains are widespread among all domains of life, yet poorly understood. Here we report the purification and characterization of an archaealJAMM/MPN+ domain protein (HvJAMM1) fromHaloferax volcaniithat cleaves ubiquitin‐like small archaeal modifier proteins (SAMP1/2) from protein conjugates.HvJAMM1 cleavedSAMP1/2 conjugates generated inH. volcaniias well as isopeptide‐ and linear‐linkedSAMP1–MoaEin purified form. Cleavage of linear linkedSAMP1–MoaEwas dependent on the presence of theSAMPdomain and theC‐terminalVSGGmotif of this domain. WhileHvJAMM1 was inhibited by size exclusion chromatography and metal chelators, its activity could be restored by addition of excessZnCl2.HvJAMM1 residues (Glu31,His88,His90,Ser98 andAsp101) that were conserved with theJAMM/MPN+ active‐site motif were required for enzyme activity. Together, these results provide the first example of aJAMM/MPN+ zinc metalloprotease that independently catalyses the cleavage of ubiquitin‐like (isopeptide and linear) bonds from target proteins. In archaea,HvJAMM1 likely regulates sampylation and the pools of ‘free’SAMPavailable for protein modification.HvJAMM1‐type proteins are thought to release theSAMPs from proteins modified post‐translationally as well as those synthesized as domain fusions.
Inflammatory histopathogenesis of nasopalatine duct cyst: a clinicopathological study of 41 cases
Authors: Tsuneki M , Maruyama S , Yamazaki M , Abé T , Adeola HA , Cheng J , Nishiyama H , Hayashi T , Kobayashi T , Takagi R , Funayama A , Saito C , Saku T
Published: September 2012

Objective

The aim of this study is to characterize immunohistochemical profiles of lining epithelia of nasopalatine duct cyst (NPC) as well as to correlate those findings with their clinicopathological features to understand the histopathogenesis of NPC.

Materials and methods

Forty‐one surgical specimens from NPC were examined for clinical profiles and expression of keratin‐7, 13, MUC‐1, and P63 by immunohistochemistry, compared to radicular cyst (RC) and maxillary sinusitis.

Results

Nasopalatine duct cyst was clinically characterized by male predominant occurrence: 44% of the cases involved tooth roots, and 70% with inflammatory backgrounds. Lining epithelia of NPCs without daughter cysts were immunohistochemically distinguished into three layers: a keratin 7‐positive (+) ciliated cell layer in the surface, a keratin‐13+ middle layer, and a MUC‐1+/P63+ lower half, indicating that they were not respiratory epithelia, and the same layering pattern was observed in RC. However, those immunolocalization patterns of the main cyst lining with daughter cyst were exactly the same as those of daughter cyst linings as well as duct epithelia of mucous glands.

Conclusions

Two possible histopathogeneses of NPC were clarified: one was inflammatory cyst like RC and the other was salivary duct cyst‐like mucocele.
An evaluation of vitamin E and pycnogenol in children suffering from oral mucositis during cancer chemotherapy
Authors: Khurana H , Pandey RK , Saksena AK , Kumar A
Published: September 2012

Objective

The purpose of the present study was to evaluate and compare the effectiveness of Vitamin E (E) and Pycnogenol (P) in treatment of Chemotherapy‐Related Oral Mucositis (ChROM) in children.

Materials and Methods

A total of 72 children, aged between 6 and 15 years, with ChROM were selected and randomly divided into three groups after assessment of oral mucositis (OM) by WHO grading system. Glycerine (control), E, and P were topically applied in group I, II, and III, respectively, and the prognosis of OM was assessed by functional, objective, and subjective parameters.

Results

Patients of group II and III showed significant improvement when ChROM was analyzed through scoring systems – WHO grading, Oral Mucositis Assessment Scale (OMAS), and Children's International Mucositis Evaluation Scale (ChIMES) as compared to group I (P < 0.001); however, there was no significant difference between groups II and III.

Conclusion

Both the drugs E and P per se are effective for treatment of OM with P being not effective for the treatment of severe mucositis (grade 4). Combination of E and P and additional agents may be tried for better results.
A randomized trial comparing the Ambu®Aura‐i™ with the air‐Q™ intubating laryngeal airway as conduits for tracheal intubation in children
Authors: Jagannathan Narasimhan , Sohn Lisa E. , Sawardekar Amod , Gordon Jason , Shah Ravi D. , Mukherji Isabella I. , Roth Andrew G. , Suresh Santhanam
Published: September 2012

Objectives

To assess the clinical performance of the Ambu Aura‐i (Aura‐i) in children.

Aim

To compare the Aura‐i with the air‐Q intubating laryngeal airway (air‐Q) for the purposes of fiberoptic‐guided tracheal intubation.

Background

The Aura‐i is a new supraglottic airway designed for tracheal intubation.

Materials/Methods

One hundred twenty children, ages 1 month to 6 years, were randomized to receive either the Aura‐i or air‐Q, and divided into three equal subgroups (Group 1, 2, 3) based on weight. The time for successful tracheal intubation was primarily assessed. The ease, time, and number of attempts for successful device insertion, leak pressures, fiberoptic grade of laryngeal view, number of attempts time for removal of the device after tracheal intubation, and complications were secondarily assessed.

Results

Device placement, tracheal intubation, and removal after tracheal intubation were successful in all patients. There were no differences in the time to successful tracheal intubation through the Aura‐i (32.9 ± 13.3 s), and the air‐Q (33.9 ± 13 s; P = 0.68), or fiberoptic grade of view between devices. There was not a statistically significant correlation between the time to intubation and the fiberoptic grade of laryngeal view in any of the groups. There were no statistically significant differences in the overall leak pressures, air‐Q (18.3 ± 6.1 cm H2O) vs Aura‐i (16 ± 5.1 cm H2O; P = 0.05). In Group 1 (5–10 kg), leak pressures were higher with the air‐Q (23.4 ± 7.2 cm H2O) than the Aura‐i (16.1 ± 5.2 cm H2O; P = 0.001). There were no statistically significant differences in the time for removal between the two devices (P = 0.11). However, with the size 1.5 Aura‐i, the pilot balloon of the tracheal tube was removed in order to facilitate the removal of the device after tracheal intubation.

Conclusions

Both devices served as effective conduits for fiberoptic‐guided tracheal intubation. The limitation of the narrower proximal airway tube of the size 1.5 Aura‐i should be considered if cuffed tracheal tubes are to be utilized.
Mitophagy or how to control the Jekyll and Hyde embedded in mitochondrial metabolism: implications for melanoma progression and drug resistance
Authors: Soengas María S.
Published: September 2012

Proteins and pathways that control cell fate are placed under intense scrutiny. The same tight regulation applies to essential organelles that can both sustain cell survival or promote self‐degradation programs. Mitochondria are perhaps the prime example of cellular machineries with split functions (personalities). As a main source of ATP, mitochondria represent the main powerhouse of eukaryotic cells. However, mitochondrial respiration has the hidden complication of the production of potentially harmful reactive oxygen species (ROS). Moreover, mitochondria holds an armamentarium of stress‐response factors, which depending on the context, may lead to pro‐inflammatory signals, and to various forms of cell death, ranging from apoptosis to necrosis. A main clearance mechanism to eliminate superfluous, damaged or hyperactive mitochondria is selective mitophagy. Mitophagy, in fact, is emerging as a key quality‐control mechanism in cancer cells. Specifically, malignant transformation has been found to induce marked changes in mitochondrial dynamics and structure. Moreover, a key hallmark of tumor progression is metabolic reprogramming, which further deregulates ROS content and renders cells more susceptible to mitochondrial perturbations. Despite its increasing relevance in cancer biology, the field of mitophagy remains virtually unexplored in melanoma. However, given unique antioxidant mechanisms in melanocytic cells (e.g., linked to melanin) and the idiosyncratic interplay between ROS and hypoxia (both mitophagy inducers) in melanoma, this tumor type represents an ideal scenario for physiological studies of mitochondrial turnover. This perspective summarizes proof of concept for in‐depth basic and translational studies of mitophagy in melanoma. Particular emphasis is dedicated to new opportunities for gene discovery and drug design in this still aggressive disease.
LBD18 acts as a transcriptional activator that directly binds to the EXPANSIN14 promoter in promoting lateral root emergence of Arabidopsis
Authors: Lee Han Woo , Kim Min‐Jung , Kim Nan Young , Lee Sung Haeng , Kim Jungmook
Published: September 2012

Lateral root formation, a developmental process under the control of the plant hormone auxin, is a major determinant of root architecture, and defines the ability of a plant to acquire nutrients and water. The LATERAL ORGAN BOUNDARIES DOMAIN/ASYMMETRIC LEAVES2‐LIKE (LBD/ASL) proteins play an important role in the lateral organ development of plants, including lateral root formation. However, their downstream components and signalling mechanisms are largely unknown. Here, we show that auxin‐responsive LBD18/ASL20 acts as a specific DNA‐binding transcriptional activator that directly regulates EXPANSIN14 (EXP14), a gene encoding a cell wall‐loosening factor that promotes lateral root emergence in Arabidopsis thaliana. We showed that LBD18 possesses transcription‐activating function in both yeast and Arabidopsis protoplasts. We isolated putative LBD18 target genes by microarray analysis, and identified EXP14 as a direct target of LBD18. Dexamethasone‐induced expression of LBD18 under the CaMV 35S promoter in transgenic Arabidopsis resulted in enhanced expression of GUS fused to the EXP14 promoter in primordium and overlaying tissues. In contrast, GUS expression under the EXP14 promoter in the lbd18 mutant background was significantly reduced in the same tissues. Experiments using a variety of molecular techniques demonstrated that LBD18 activates EXP14 by directly binding to a specific promoter element in vitro and in vivo. Overexpression of EXP14 in Arabidopsis resulted in the stimulation of emerged lateral roots, but not primordia, whereas EXP14 loss‐of‐function plants had reduced auxin‐stimulated lateral root formation. This study revealed the molecular function of LBD18 as a specific DNA‐binding transcription factor that activates EXP14 expression by directly binding to its promoter.
Activation of R‐mediated innate immunity and disease susceptibility is affected by mutations in a cytosolic O‐acetylserine (thiol) lyase in Arabidopsis
Authors: Tahir Jibran , Watanabe Mutsumi , Jing Hai‐Chun , Hunter Donald A. , Tohge Takayuki , Nunes‐Nesi Adriano , Brotman Yariv , Fernie Alisdair R. , Hoefgen Rainer , Dijkwel Paul P.
Published: September 2012

O‐acetylserine (thiol) lyases (OASTLs) are evolutionarily conserved proteins among many prokaryotes and eukaryotes that perform sulfur acquisition and synthesis of cysteine. A mutation in the cytosolic OASTL‐A1 protein ONSET OF LEAF DEATH3 (OLD3) was previously shown to reduce the OASTL activity of the old3‐1 protein in vitro and cause auto‐necrosis in specific Arabidopsis accessions. Here we investigated why a mutation in this protein causes auto‐necrosis in some but not other accessions. The auto‐necrosis was found to depend on Recognition of Peronospora Parasitica 1 (RPP1)‐like disease resistance R gene(s) from an evolutionarily divergent R gene cluster that is present in Ler‐0 but not the reference accession Col‐0. RPP1‐like gene(s) show a negative epistatic interaction with the old3‐1 mutation that is not linked to reduced cysteine biosynthesis. Metabolic profiling and transcriptional analysis further indicate that an effector triggered‐like immune response and metabolic disorder are associated with auto‐necrosis in old3‐1 mutants, probably activated by an RPP1‐like gene. However, the old3‐1 protein in itself results in largely neutral changes in primary plant metabolism, stress defence and immune responses. Finally, we showed that lack of a functional OASTL‐A1 results in enhanced disease susceptibility against infection with virulent and non‐virulent Pseudomonas syringae pv. tomato DC3000 strains. These results reveal an interaction between the cytosolic OASTL and components of plant immunity.
A proteomics approach to investigate the process of Zn hyperaccumulation in Noccaea caerulescens (J & C. Presl) F.K. Meyer
Authors: Schneider Thomas , Persson Daniel Pergament , Husted Søren , Schellenberg Maja , Gehrig Peter , Lee Youngsook , Martinoia Enrico , Schjoerring Jan K. , Meyer Stefan
Published: September 2012

Zinc (Zn) is an essential trace element in all living organisms, but is toxic in excess. Several plant species are able to accumulate Zn at extraordinarily high concentrations in the leaf epidermis without showing any toxicity symptoms. However, the molecular mechanisms of this phenomenon are still poorly understood. A state‐of‐the‐art quantitative 2D liquid chromatography/tandem mass spectrometry (2D‐LC‐MS/MS) proteomics approach was used to investigate the abundance of proteins involved in Zn hyperaccumulation in leaf epidermal and mesophyll tissues of Noccaea caerulescens. Furthermore, the Zn speciation in planta was analyzed by a size‐exclusion chromatography/inductively coupled plasma mass spectrometer (SEC‐ICP‐MS) method, in order to identify the Zn‐binding ligands and mechanisms responsible for Zn hyperaccumulation. Epidermal cells have an increased capability to cope with the oxidative stress that results from excess Zn, as indicated by a higher abundance of glutathione S‐transferase proteins. A Zn importer of the ZIP family was more abundant in the epidermal tissue than in the mesophyll tissue, but the vacuolar Zn transporter MTP1 was equally distributed. Almost all of the Zn located in the mesophyll was stored as Zn–nicotianamine complexes. In contrast, a much lower proportion of the Zn was found as Zn–nicotianamine complexes in the epidermis. However, these cells have higher concentrations of malate and citrate, and these organic acids are probably responsible for complexation of most epidermal Zn. Here we provide evidence for a cell type‐specific adaptation to excess Zn conditions and an increased ability to transport Zn into the epidermal vacuoles.
Arabidopsis RABA1 GTPases are involved in transport between the trans‐Golgi network and the plasma membrane, and are required for salinity stress tolerance
Authors: Asaoka Rin , Uemura Tomohiro , Ito Jun , Fujimoto Masaru , Ito Emi , Ueda Takashi , Nakano Akihiko
Published: September 2012

RAB GTPases are key regulators of membrane traffic. Among them, RAB11, a widely conserved sub‐group, has evolved in a unique way in plants; plant RAB11 members show notable diversity, whereas yeast and animals have only a few RAB11 members. Fifty‐seven RAB GTPases are encoded in the Arabidopsis thaliana genome, 26 of which are classified in the RAB11 group (further divided into RABA1–RABA6 sub‐groups). Although several plant RAB11 members have been shown to play pivotal roles in plant‐unique developmental processes, including cytokinesis and tip growth, molecular and physiological functions of the majority of RAB11 members remain unknown. To reveal precise functions of plant RAB11, we investigated the subcellular localization and dynamics of the largest sub‐group of Arabidopsis RAB11, RABA1, which has nine members. RABA1 members reside on mobile punctate structures adjacent to the trans‐Golgi network and co‐localized with VAMP721/722, R‐SNARE proteins that operate in the secretory pathway. In addition, the constitutive‐active mutant of RABA1b, RABA1bQ72L , was present on the plasma membrane. The RABA1b ‐containing membrane structures showed actin‐dependent dynamic motion . Vesicles labeled by GFP–RABA1b moved dynamically, forming queues along actin filaments. Interestingly, Arabidopsis plants whose four major RABA1 members were knocked out, and those expressing the dominant‐negative mutant of RABA1B, exhibited hypersensitivity to salinity stress. Altogether, these results indicate that RABA1 members mediate transport between the trans‐Golgi network and the plasma membrane, and are required for salinity stress tolerance.
Variability in the Distribution of Genes Encoding Virulence Factors and Putative Extracellular Proteins of Streptococcus pyogenes in India, a Region with High Streptococcal Disease Burden, and Implication for Development of a Regional Multisubunit Vaccine
Authors: Sagar Vivek , Bergmann René , Nerlich Andreas , McMillan David J. , Nitsche Schmitz D. Patric , Chhatwal Gursharan S.
Published: September 2012

Streptococcus pyogenescauses a wide variety of human diseases and is a significant cause of morbidity and mortality. Attempts to develop a vaccine were hampered by the genetic diversity ofS. pyogenesacross different regions of the world. This study sought to identify streptococcal antigens suitable for a region-specific vaccine in India. We used a two-step approach, first performing epidemiological analysis to identify the conserved antigens among Indian isolates. The second step consisted of validating the identified antigens by serological analysis. The 201 streptococcal clinical isolates from India used in this study represented 69 differentemmtypes, withemm12 being the most prevalent. Virulence profiling of the North and South IndianS. pyogenesisolates with a custom-designed streptococcal virulence microarray identified seven conserved putative vaccine candidates. Collagen-like surface protein (SCI), putative secreted 5′-nucleotidase (PSNT), and C5a peptidase were found in 100% of the isolates, while R28, a putative surface antigen (PSA), and a hypothetical protein (HYP) were found in 90% of the isolates. A fibronectin binding protein, SfbI, was present in only 78% of the isolates. In order to validate the identified potential vaccine candidates, 185 serum samples obtained from patients with different clinical manifestations were tested for antibodies. Irrespective of clinical manifestations, serum samples showed high antibody titers to all proteins except for SCI and R28. Thus, the data indicate that PSNT, C5a peptidase, PSA, HYP, and SfbI are promising candidates for a region-specific streptococcal vaccine for the different parts of India.
Western Blotting Is an Efficient Tool for Differential Diagnosis of Paracoccidioidomycosis and Pulmonary Tuberculosis
Authors: Bertoni Thâmara Aline , Perenha-Viana Maysa Cláudia Zolin , Patussi Eliana Valéria , Cardoso Rosilene Fressatti , Svidzinski Terezinha Inez Estivalet
Published: September 2012

Sputum and sera from 134 patients screened for tuberculosis (TB) were analyzed to investigate TB and paracoccidioidomycosis (PCM). Of these patients, 11 (8.2%) were confirmed to have TB, but six (4.5%) were positive only for PCM. All patients with PCM presented anti-43-kDa-component antibodies in Western blotting (WB) assays, while in the TB-positive patients these antibodies did not appear. This preliminary study suggests WB as a potential tool for differential laboratory diagnosis between TB and PCM.
Misdiagnosis of Late-Onset Lyme Arthritis by Inappropriate Use of Borrelia burgdorferi Immunoblot Testing with Synovial Fluid
Authors: Barclay Sam S. , Melia Michael T. , Auwaerter Paul G.
Published: September 2012

The primary objective of this study was to determine whether patients with putative late-onset Lyme arthritis based upon synovial fluidBorrelia burgdorferiIgM and IgG immunoblot testing offered by commercial laboratories satisfied conventional criteria for the diagnosis of Lyme arthritis. Secondary objectives included assessing the prior duration and responsiveness of associated antibiotic therapy. We conducted a retrospective analysis of 11 patients referred to an academic medical center infectious disease clinic during the years 2007 to 2009 with a diagnosis of Lyme disease based upon previously obtained synovial fluidB. burgdorferiimmunoblot testing. Ten of the 11 (91%) patients with a diagnosis of late-onset Lyme arthritis based upon interpretation of synovial fluidB. burgdorferiimmunoblot testing were seronegative and did not satisfy published criteria for the diagnosis of late-onset Lyme arthritis. None of the 10 patients had a clinical response to previously received antibiotics despite an average course of 72 days. Diagnosis of Lyme arthritis should not be based on synovial fluidB. burgdorferiimmunoblot testing. This unvalidated test does not appear useful for the diagnosis of Lyme disease, and this study reinforces the longstanding recommendation to useB. burgdorferiimmunoblot testing only on serum samples and not other body fluids. Erroneous interpretations of “positive” synovial fluid immunoblots may lead to inappropriate antibiotic courses and delays in diagnosis of other joint diseases.
Measles Virus IgG Avidity Assay for Use in Classification of Measles Vaccine Failure in Measles Elimination Settings
Authors: Mercader Sara , Garcia Philip , Bellini William J.
Published: September 2012

In regions where endemic measles virus has been eliminated, diagnostic assays are needed to assist in correctly classifying measles cases irrespective of vaccination status. A measles IgG avidity assay was configured using a commercially available measles-specific IgG enzyme immunoassay by modifying the protocol to include three 5-min washes with diethylamine (60 mM; pH 10.25) following serum incubation; serum was serially diluted, and the results were expressed as the end titer avidity index. Receiver operating characteristic analysis was used for evaluation and validation and to establish low (≤30%) and high (≥70%) end titer avidity thresholds. Analysis of 319 serum specimens expected to contain either high- or low-avidity antibodies according to clinical and epidemiological data indicated that the assay is highly accurate, with an area under the curve of 0.998 (95% confidence interval [CI], 0.978 to 1.000), sensitivity of 91.9% (95% CI, 83.2% to 97.0%), and specificity of 98.4% (95% CI, 91.6% to 100%). The assay is rapid (<2 h) and precise (standard deviation [SD], 4% to 7%). In 18 samples from an elimination setting outbreak, the assay identified 2 acute measles cases with low-avidity results; both were IgM-positive samples. Additionally, 11 patients (15 samples) with modified measles who were found to have high-avidity IgG results were classified as secondary vaccine failures; one sample with an intermediate-avidity result was not interpretable. In elimination settings, measles IgG avidity assays can complement existing diagnostic tools in confirming unvaccinated acute cases and, in conjunction with adequate clinical and epidemiologic investigation, aid in the classification of vaccine failure cases.
Two Outer Membrane Lipoproteins from Histophilus somni Are Immunogenic in Rabbits and Sheep and Induce Protection against Bacterial Challenge in Mice
Authors: Guzmán-Brambila Carolina , Rojas-Mayorquín Argelia E. , Flores-Samaniego Beatriz , Ortuño-Sahagún Daniel
Published: September 2012

Histophilus somniis an economically important pathogen of cattle and other ruminants and is considered one of the key components of the bovine respiratory disease (BRD) complex, the leading cause of economic loss in the livestock industry. BRD is a multifactorial syndrome, in which a triad of agents, including bacteria, viruses, and predisposing factors or “stressors,” combines to induce disease. Although vaccines againstH. somnihave been used for many decades, traditional bacterins have failed to demonstrate effective protection in vaccinated animals. Hence, the BRD complex continues to produce strong adverse effects on the health and well-being of stock and feeder cattle. The generation of recombinant proteins may facilitate the development of more effective vaccines againstH. somni, which could confer better protection against BRD. In the present study, primers were designed to amplify, clone, express, and purify two recombinant lipoproteins fromH. somni, p31 (Plp4) and p40 (LppB), which are structural proteins of the outer bacterial membrane. The results presented here demonstrate, to our knowledge for the first time, that when formulated, an experimental vaccine enriched with these two recombinant lipoproteins generates high antibody titers in rabbits and sheep and exerts a protective effect in mice against septicemia induced byH. somnibacterial challenge.
Clonal Analysis of Neisseria meningitidis Serogroup B Strains in South Africa, 2002 to 2006: Emergence of New Clone ST-4240/6688
Authors: Moodley Chivonne , du Plessis Mignon , Ndlangisa Kedibone , de Gouveia Linda , Klugman Keith P. , von Gottberg Anne ,   for GERMS-SA (Group for Enteric, Respiratory and Meningeal Disease Surveillance in South Africa)
Published: September 2012

From August 1999 through July 2002, hyperinvasive
Neisseria meningitidis
serogroup B (MenB) clonal complexes (CCs), namely, ST-32/ET-5 (CC32) and ST-41/44/lineage 3 (CC41/44), were predominant in the Western Cape Province of South Africa. This study analyzed MenB invasive isolates from a national laboratory-based surveillance system that were collected from January 2002 through December 2006. Isolates were characterized by pulsed-field gel electrophoresis (PFGE) (
n
= 302), and multilocus sequence typing (MLST) and PorA and FetA typing were performed on randomly selected isolates (34/302, 11%). In total, 2,400 cases were reported, with the highest numbers from Gauteng Province (1,307/2,400, 54%) and Western Cape Province (393/2,400, 16%); 67% (1,617/2,400) had viable isolates and 19% (307/1,617) were identified as serogroup B. MenB incidence remained stable over time (
P
= 0.77) (average incidence, 0.13/100,000 population [range, 0.10 to 0.16/100,000 population]). PFGE (302/307, 98%) divided isolates (206/302, 68%) into 13 clusters and 96 outliers. The largest cluster, B1, accounted for 25% of isolates (76/302) over the study period; its prevalence decreased from 43% (20/47) in 2002 to 13% (8/62) in 2006 (
P
< 0.001), and it was common in the Western Cape (58/76, 76%). Clusters B2 and B3 accounted for 10% (31/302) and 6% (19/302), respectively, and showed no significant change over time and were predominant in Gauteng. Randomly selected isolates from clusters B1, B2, and B3 belonged to CC32, CC41/44, and the new CC4240/6688, respectively. Overall, 15 PorA and 12 FetA types were identified. MenB isolates were mostly diverse with no single dominant clone; however, CC32 and CC41/44 accounted for 35% and the new CC4240/6688 was the third most prevalent clone.
Molecular Typing of Treponema pallidum: a 5-Year Surveillance in Shanghai, China
Authors: Dai Ting , Li Kang , Lu Haikong , Gu Xin , Wang Qianqiu , Zhou Pingyu
Published: September 2012

Previously, a small study showed that 14f was the predominant subtype of
Treponema pallidum
in Shanghai, China. The result was quite different from the genotype distribution in other areas of China. This study aimed to identify the strain types of
Treponema pallidum
in samples collected over a 5-year period in Shanghai. From 2007 to 2011, genital swabs were collected from patients with syphilis from the Shanghai Skin Disease Hospital. Positive specimens were typed by the enhanced typing method by adding a
tp0548
gene to the existing
arp
and
tpr
genotype system. In total, 304 of the 372 enrolled patients yielded fully typeable DNA. Ten
arp
types (4, 6, 8, 9, 11, 12, 13, 14, 15, and 19), 3
tpr
types (a, d, and o), and 5
tp0548
types (a, c, f, g, and i) were identified. In total, 12 subtypes were identified with a combination of the
arp
and
tpr
genes. Subtype 14d was found in 270 samples (88.8%). When the combination included the
tp0548
gene, the 12 CDC subtypes identified were divided into 14 strain types. The predominant type was 14d/f (88.8%), followed by 15d/f (3.6%), 13d/f (1.3%), and 19d/c (1.3%). Two of the 44 14d/f-infected patients and both of the 19d/c-infected patients who underwent a lumbar puncture were diagnosed with neurosyphilis. This study showed that the predominant type in Shanghai was 14d/f. While this is in keeping with data from other areas in China, it is different from an earlier report showing that 14f is the most common genotype in Shanghai. Further studies are needed to better understand the association between strain types and neurosyphilis.