bluuebunny/crossref_metadata_2025 huggingface dataset:
Coronary artery disease is associated with higher epicardial Retinol‐binding protein 4 (RBP4) and lower glucose transporter (GLUT) 4 levels in epicardial and subcutaneous adipose tissue
Authors: Salgado‐Somoza Antonio , Teijeira‐Fernández Elvis , Rubio José , Couso Elena , González‐Juanatey José R. , Eiras Sonia
Published: June 2011

Objective  Retinol‐binding protein 4 (RBP4), produced by adipocytes and hepatocytes, contributes to an unfavourable lipid profile and insulin resistance, which can contribute to the development of coronary artery disease (CAD). Recently, several studies have shown that epicardial adipose tissue (EAT) differs from subcutaneous adipose tissue (SAT) and plays a role on the physiopathology of CAD because of its proximity to the coronary arteries. We aimed to study the expression and secretion levels of RBP4 in both fat tissues and explore its possible association with CAD.

Research Design and Methods  Fifty‐eight patients undergoing heart surgery were included in the study. We analysed RBP4 mRNA expression by real‐time PCR, protein expression by Western blot and immunohistochemistry, and secretion of EAT and SAT explants from CAD and non‐CAD patients by Enzyme Immunoassay.

Results  Retinol‐binding protein 4 is expressed at similar levels in EAT and SAT, mainly from adipocytes. Protein levels were higher in EAT from CAD than non‐CAD patients (0·63 ± 0·09 arbitrary units (a.u).; n = 10) vs (0·41 ± 0·04 a.u.; n = 13, P = 0·039). In contrast, GLUT4 mRNA levels were lower in EAT from CAD than non‐CAD patients (6·55 ± 0·16 a.u.; n = 13) vs (7·21 ± 0·18 a.u.; n = 14, P = 0·012). We also found differential expression in SAT between samples from CAD and non‐CAD patients [(6·63 ± 0·16 a.u.; n = 14) vs (7·21 ± 0·14 a.u.; n = 14, P = 0·009)]. Besides, EAT releases higher RBP4 levels than SAT after 3, 6, 24 and 48 h of culture. These levels were independent of CAD but significantly higher in diabetic than nondiabetic patients.

Conclusion  Retinol‐binding protein 4 levels behave differently in EAT and SAT with respect to CAD. However, both adipose tissues have lower GLUT4 levels in patients with CAD. These findings suggest a differential regulation of RBP4 production in EAT and SAT that may be influenced by local factors.
Atmospheric nitrogen deposition explains patterns of plant species loss
Authors: McCLEAN COLIN J. , van den BERG LEON J. L. , ASHMORE MIKE R. , PRESTON CHRIS D.
Published: June 2011

Atmospheric nitrogen (N) deposition across Europe increased substantially from the 1950s to the 1990s. Targeted surveys suggest a negative correlation between N deposition and species richness within quadrats in sensitive habitats. However, it remains unclear whether plant species losses at national recording scales are correlated with nitrogen deposition. We relate plant species losses before 1987 in Great Britain to reduced and oxidized N deposition, land use change and climate change. The mean Ellenberg fertility (N) indices of plant species lost in each 100 km2 cell before 1987 was compared with those of species that were recorded between 1987 and 1999. In 45% of squares, indices of species lost were significantly lower than those for species present after 1986. For 17%, primarily upland, squares, the opposite effect was found. A generalized least squares regression model, with difference in the mean Ellenberg N index between samples as the dependent variable, showed that higher deposition of reduced N was significantly associated with selective loss of species with a lower index. Arable land use and change in arable land use also demonstrated this positive relationship. Rough grazing, change in rough grazing, change in pasture and change in annual precipitation showed negative effects. Difference in Ellenberg R index was highly correlated with difference in Ellenberg N and was negatively correlated with oxidized N deposition, suggesting that the lack of a significant effect of oxidized N deposition on Ellenberg N was because it had effects through both acidification and eutrophication, while the effect of reduced N deposition was primarily through eutrophication. Our results suggest that N deposition, along with land use and precipitation changes, has been a significant driver of local plant extinctions. With N deposition increasing in many parts of the world, local extinctions of plant species may be experienced in other regions.
Methane emissions from sheep pasture, measured with an open‐path eddy covariance system
Authors: Dengel Sigrid , Levy Peter E. , Grace John , Jones Stephanie K. , Skiba Ute M.
Published: June 2011

Methane (CH4) is an important greenhouse gas, contributing 0.4–0.5 W m−2 to global warming. Methane emissions originate from several sources, including wetlands, rice paddies, termites and ruminating animals. Previous measurements of methane flux from farm animals have been carried out on animals in unnatural conditions, in laboratory chambers or fitted with cumbersome masks. This study introduces eddy covariance measurements of CH4, using the newly developed LI‐COR LI‐7700 open‐path methane analyser, to measure field‐scale fluxes from sheep grazing freely on pasture. Under summer conditions, fluxes of methane in the morning averaged 30 nmol m−2 s−1, whereas those in the afternoon were above 100 nmol m−2 s−1, and were roughly two orders of magnitude larger than the small methane emissions from the soil. Methane emissions showed no clear relationship with air temperature or photosynthetically active radiation, but some diurnal pattern was apparent, probably linked to sheep grazing behaviour and metabolism. Over the measurement period (days 60–277, year 2010), cumulative methane fluxes were 0.34 mol CH4 m−2, equating to 134.3 g CO2 equivalents m−2. By comparison, a carbon dioxide (CO2) sink of 819 g CO2 equivalents m−2 was measured over the same period, but it is likely that much of this would be released back to the atmosphere during the winter or as off‐site losses (through microbial and animal respiration). By dividing methane fluxes by the number of sheep in the field each day, we calculated CH4 emissions per head of livestock as 7.4 kg CH4 sheep−1 yr−1, close to the published IPCC emission factor of 8 kg CH4 sheep−1 yr−1.
Molecular diagnosis of haemophilia A at Centro Hospitalar de Coimbra in Portugal: study of 103 families – 15 new mutations
Authors: SILVA PINTO C. , FIDALGO T. , SALVADO R. , MARQUES D. , GONÇALVES E. , MARTINHO P. , MARKOFF A. , MARTINS N. , LETÍCIA RIBEIRO M.
Published: June 2011

Summary.  Haemophilia A (HA), the most commonly inherited bleeding disorder, has well known phenotype heterogeneity, influenced by the type of mutation, modulating factors and development of inhibitors. Nowadays, new technologies in association with bioinformatics tools allow a better genotype/phenotype correlation. With the main objective of identifying familial carrier women and to offer prenatal diagnosis, 141 HA patients belonging to 103 families, followed or referred to the Haemophilia Centre of CHC, E.P.E., were studied. Molecular diagnosis strategy was based on HA severity: IVS22 and IVS1 inversions, direct sequencing and MLPA technique. New missense and splicing mutations were further analyzed using molecular modelling. Genotype/phenotype correlation was assessed taking into account the known modulating factors. During this study, mutations were detected in 102/103 families, carrier status was determined in 83 women and 14 prenatal diagnoses were performed. In a total of 46 different mutations identified, 15 have not been reported previously by the HAMSTeRS and HGMD. Genotype/phenotype correlation revealed two cases with a clinical picture less severe than expected by the type of mutation identified. Six patients developed inhibitors: five severe (IVS22, IVS1, large deletion) and one mild (p. Gln2265Lys). The adopted strategy allowed the identification of 99% of the molecular alterations underlying the HA phenotype (98% detection rate for severe and 100% for moderate and mild). Evaluation of genotype–phenotype correlation was complemented with structural protein modelling of newly identified missense mutations, contributing to better understanding of the disease‐causing mechanisms and to deepening knowledge on protein structure‐function.
The contribution of high pressure carbon dioxide in the inactivation kinetics and structural alteration of myrosinase
Authors: Yang Yingjie , Liao Xiaojun , Hu Xiaosong , Zhang Yan , Chen Fang , Wu Jihong
Published: June 2011

High pressure carbon dioxide (HPCD) has been verified to be an efficient way of inactivating enzyme activity. This work investigates the influence of temperature (T), pressure (P), exposure times (t) on the activity of commercial myrosinase (MYR) submitted to HPCD. Results showed that only 1.00% of MYR activity retained at 22 MPa and 65 °C for 5 min. Moreover, the first‐order reaction kinetic data of MYR inactivation as influenced by pressure of HPCD were analysed. With the pressure rising from 8 to 22 MPa at 55 °C, the inactivation rate constant (k) increased from 0.015 to 0.024 min−1, while the decimal reduction time (D) decreased from 157.2 to 96.1 min. Additionally, a series of exploratory experiments were conducted to investigate the contribution of the HPCD parameters (T, P and CO2 dissolution), with analysing circular dichroism spectroscopy and tryptophan fluorescence spectra, illustrate that CO2 dissolution plays a dominant role in MYR inactivation and structural alteration.
Effect of heat treatment on the properties of soy protein‐stabilised emulsions
Authors: Li Fang , Kong Xiangzhen , Zhang Caimeng , Hua Yufei
Published: June 2011

The effect of heat treatment on the properties of soy protein‐stabilised emulsions was investigated. Emulsions were prepared with unheated and heat‐treated soy protein (NSP and HSP) dispersions. Heating on soy protein dispersions at 95 °C for 30 min resulted in smaller average oil droplet size, lower tendency for oil droplet flocculation, higher protein adsorption and lower viscosity. The properties of emulsions were significantly influenced by the protein concentration. The sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) profiles showed that the heat treatment on soy protein dispersions increased the protein adsorption at O/W interface. The viscosity of all samples at low shear rate was inversely proportional to the d32, suggesting a positive relation to the total interfacial area per unit volume. Emulsions showed shear‐thinning behaviour. The relaxation time was found to increase with aqueous phase viscosity determined by the Cross viscosity model.
Influence of tragacanth gum exudates from specie of Astragalus gossypinus on rheological and physical properties of whey protein isolate stabilised emulsions
Authors: Samavati Vahid , Emam‐Djomeh Zahra , Mohammadifar Mohammad Amin , Omid Mahmoud , Mehdinia Ali
Published: June 2011

The aim of this work was to investigate the effects of Iranian tragacanth gum (Astragalus gossypinus) (0.5, 1 wt.%), Whey protein isolate (WPI) (2, 4 wt.%) and acid oleic‐phase volume fraction (5, 10% v/v) on droplet size distribution, creaming index and rheological properties of emulsions with various compositions. Rheological investigations showed that both loss and storage modules increased with gum and oil contents. However, the viscoelastic behaviour was mainly governed by the gum concentration. Delta degree (storage and loss modules ratio) increased with frequency indicated that liquid like viscose behaviour dominates over solid like elastic behaviour. The shear‐thinning behaviour of all dispersions was successfully modelled with power law and Ellis models and Ellis model was founded as the better model to describe the flow behaviour of dispersions. Droplet size distribution was measured by light scattering; microscopic observations revealed a flocculated system. Increase in gum, WPI and oil contents resulted in decrease in creaming index of emulsions with dominant effect of gum concentration.
Effect of ageing on different egg yolk fractions on surface properties at the air–water interface
Authors: Freschi Jérôme , Razafindralambo Hary , Danthine Sabine , Blecker Christophe
Published: June 2011

The aim of this study was to evaluate the impact of egg ageing on the surface properties of whole and fractionated yolk at the air–water interface. Eggs were stored at 4°C for 24 h, 1 week and 3 weeks after laying. A laboratory scale fractionation process was then applied at each ageing time. Egg yolk was separated into two fractions, plasma composed of low‐density lipoproteins (LDLs) and livetins, and granules formed by high‐density lipoproteins (HDLs), phosvitins and LDLg (g  =  granule). Moreover, recombined plasma and granules fractions were investigated to highlight a potential synergic effect on surface properties. Results have shown the main contribution of LDLs on surface properties of yolk and an improvement of granules surface properties when they are disrupted. Moreover, ageing affected surface properties differently depending on the considered fractions. Broken LDLs and disrupted granules could explain this observed behaviour. Recombined fractions showed different compression isotherms at the air–water interface than whole yolk.
Iron content and availability studies in some Sri Lankan rice varieties
Authors: Herath Herath M. T. , Rajapakse Damitha , Wimalasena Sukumal , Weerasooriya M. K. Bandu
Published: June 2011

Thirty‐eight rice varieties, grown in Low Country Wet Zone, during Yala (2006) and Maha (2006/2007) seasons were screened for stable high iron rice varieties and were statistically analysed. Iron contents ranged 1.89–3.73 mg 100 g−1 and varied significantly (P < 0.05) with variety and season. Selected fifteen high iron rice varieties, at degree of polishing 8–10% were analysed for iron, zinc and phytate. High iron contents in endosperm were observed in Suduru Samba (0.47 mg 100 g−1), Basmati 370 (0.37 mg 100 g−1), Kalu Heenati (0.42 mg 100 g−1), Rathu Heenati (0.44 mg 100 g−1) and Sudu Heenati (0.37 mg 100 g−1). Phytate contents of polished rice varieties ranged from 200–300 mg 100 g−1. Large reduction in iron content (84.5–93.6%) was observed in polished rice while the reduction in phytate content (18.9–40.8%) was low. Percentage dialyzability of iron in selected endosperm high iron rice varieties ranged from 1.73 to 8.71. Dialysability of iron in cooked polished rice did not show a relation to the phytate content in raw rice.
Recycling preservation solutions in black ripe olive processing
Authors: Medina Eduardo , García Pedro , Romero Concepción , Brenes Manuel
Published: June 2011

The purpose of this study was to evaluate the reuse of the liquid employed to preserve black ripe olives in a new fermentation process. The fermentation of olives in these reused solutions (ozonated, acetic acid distillate and old storage solutions) produced similar results as with a fresh acetic acid solution. The pH was between 3.8 and 4.1, and the free acidity ranged from 1.4 to 2.6. Yeasts and acetic acid bacteria were the predominant microorganisms, while lactic acid bacteria grew occasionally after 6–8 months of storage. Off‐odours, fruit defects or any other spoilage were not detected in any case. Moreover, the quality of the darkened product (colour, firmness, flavour) was statistically similar for all treatments, although fruits preserved in a reused ozonated liquid showed less dark fruits (R700 = 9.7–10.7) than the others (R700 = 7.7–9.3). These results open the possibility of reusing the black olive solutions in a new preservation process.
Lysyl oxidase from jumbo squid (Dosidicus gigas) muscle: detection and partial purification
Authors: Torres‐Arreola Wilfrido , Ezquerra‐Brauer Josafat M. , Figueroa‐Soto Ciria G. , Valenzuela‐Soto Elisa M. , Garcia‐Sanchez Guillermina , Marquez‐Rios Enrique , Pacheco‐Aguilar Ramón
Published: June 2011

Lysyl oxidase (LOX) was detected and partially purified from jumbo squid (Dosidicus gigas) muscle, for the first time. A procedure for the purification of LOX from jumbo squid muscle which consisted of urea extraction, Sephadex G‐75 and anion exchange chromatography was developed. Activity of partially purified LOX was 390‐fold higher than the original extract. Two protein fractions with 32 and 24 kDa were detected by SDS‐PAGE. The enzyme was strongly inhibited by β‐aminopropionitrile fumarate, a specific LOX inhibitor. LOX was purified with 3.8% yield, showing a specific activity of 0.078 IU mg−1 protein. This knowledge will help understand the behaviour of jumbo squid protein during cool storage or manufacture.
Sterolic composition and triacylglycerols of Oueslati virgin olive oil: comparison among different geographic areas
Authors: Ouni Youssef , Flamini Guido , Youssef Nabil Ben , Guerfel Mokhtar , Zarrouk Mokhtar
Published: June 2011

The main triglycerides (TG) identified in the Oueslati virgin olive oil were 1,2,3‐trioleylglycerol (OOO), 2,3‐dioleyl‐1‐palmitoylglycerol (POO) and 2,3‐dioleyl‐1‐linoleylglycerol (LOO) representing more than 80% of the total area of peaks in the chromatogram. Other minor triacylglycerols were 2,3‐dioleyl‐1‐stearoylglycerol (SOO), 2‐oleyl‐3‐palmitoyl‐1‐stearoylglycerol (SOP), 1‐linolenoyl‐2‐oleyl‐3‐palmitoylglycerol (LnOP), 1, 2‐dilinoleyl‐3‐palmitoylglycerol (LLP), 1, 3‐dioleyl‐2‐linolenoylglycerol (OLnO), 1‐linolenoyl‐2‐linoleyl‐3‐oleylglycerol (LnLO) and 1,2,3‐trilinoleylglycerol (LLL). The sterol profile of Tunisian virgin olive oils produced from Oueslati cultivar was established by gas chromatography using a flame ionisation detector (GC‐MS). More than 10 compounds were identified and characterised. As expected for virgin olive oil, the main sterols found in all Oueslati olive oils were β‐sitosterol, Δ5‐avenasterol, campesterol and stigmasterol. Cholesterol, 24‐methylenecholesterol, clerosterol, campestanol, sitostanol, Δ7‐stigmastenol, Δ5, 24‐stigmastadienol and Δ7‐avenasterol were also found in all samples, but in lower amounts. Most of these compounds are significantly affected by the geographical origin of the oil. Besides the sterol components, two triterpene dialcohols, erythrodiol and uvaol, were also detected.
Recovery and fractionation of major apple and grape polyphenols from model solutions and crude plant extracts using ion exchange and adsorbent resins
Authors: Kammerer Judith , Schweizer Christian , Carle Reinhold , Kammerer Dietmar R.
Published: June 2011

Model systems composed of phenolic compounds, amino acids and mixtures thereof mimicking apple pomace and grape skin extracts were used to examine adsorption and ion exchange behaviour of one adsorbent as well as four anion and four cation exchange resins in a column system at pH 1, 5 and 7. Furthermore, apple pomace and grape skin extracts were loaded onto the resins to evaluate the transferability of the results obtained in the model experiments. In addition, different eluents were applied for the successive recovery of phenolic compounds from the resins. The results provide valuable information on highly complex adsorption and exchange systems. The data indicate both enhanced and decreased polyphenol binding rates as a result of amino acid addition depending on the composition of the solutions applied to the resins. The application of appropriate process parameters during adsorption and desorption was shown to allow selective recovery of compound classes or even individual compounds. As the model experiments did not allow complete prediction of polyphenol binding behaviour, further extract compounds not investigated in the present study presumably also interfere with polyphenol recovery. The results help to systematically optimise industrial processes, thus improving cost‐efficiency and selectivity of polyphenol recovery.
Optimisation of formulations and infrared–microwave combination baking conditions of chestnut–rice breads
Authors: Demirkesen Ilkem , Sumnu Gulum , Sahin Serpil , Uysal Nalan
Published: June 2011

The main objective of this study was to design gluten‐free breads containing chestnut and rice flour and xanthan–guar gum blend to be baked in infrared–microwave combination oven. Response surface methodology (RSM) was used to optimise gluten‐free bread formulations and processing conditions. Weight loss, firmness, specific volume and colour change of the breads were determined. Rice flour mixed with different proportions of chestnut flour and different emulsifier contents were used to prepare breads. The gluten‐free formulations were baked using different upper halogen lamp powers, microwave powers and baking time which were varied from 40% to 80%, 30% to 70% and 9 to 17 min, respectively. Gluten‐free breads and wheat breads baked in conventional oven were used for comparison. Breads containing 46.5% chestnut flour and 0.62% emulsifier and baked using 40% infrared and 30% microwave power for 9 min had statistically comparable quality with conventionally baked ones.
Microencapsulation of Bifidobacterium bifidum F‐35 in reinforced alginate microspheres prepared by emulsification/internal gelation
Authors: Zou Qiang , Zhao Jianxin , Liu Xiaoming , Tian Fengwei , Zhang He‐ping , Zhang Hao , Chen Wei
Published: June 2011

Alginate microspheres containing Bifidobacterium bifidum F‐35 prepared by emulsification/internal gelation were reinforced by blending with pectin or starch or coating with chitosan or poly‐L‐lysine to provide extra protection for the strain. The influence of these treatments on the size of microspheres, encapsulation yield (EY) and protective effect of microencapsulation on the cells was studied. No difference was detected in EY with different types of reinforcement, which was approximately 43–50%. The mean diameter of reinforced alginate microspheres ranged from 117 to 178 μm, reaching a maximum value when starch was incorporated in the alginate matrix. It was observed that the protective effects varied with the type of reinforcement. However, chitosan‐coated alginate microspheres provided the best protection for microencapsulated cells in simulated gastrointestinal tract and during 1 month of storage at 4 °C, and this system could be the comparatively effective vector of bifidobacteria for intestinal delivery.
Plant diversity partitioning in grazed Mediterranean grassland at multiple spatial and temporal scales
Authors: Golodets Carly , Kigel Jaime , Sternberg Marcelo
Published: June 2011

1. Grazing by large ungulates may affect plant species richness and diversity at multiple spatial and/or temporal scales, because grazing affects small‐scale resource heterogeneity and plant interactions at the local scale, while effects at the landscape scale are related to grazing intensity and timing.

2. We used diversity partitioning to analyse long‐ and short‐term effects of cattle grazing on plant species richness and diversity in an experimental spatial hierarchy in Mediterranean annual grassland. Short‐term changes during secondary succession in grazed plots (2003–2005) at two grazing intensities (heavy and moderate) were analysed and compared with long‐term protected vegetation. We applied Hill’s q‐diversity metrics at q = 0 (species richness) and q = 2 (reciprocal Simpson diversity) to examine the partitioning of species richness and diversity between their alpha (α) and beta (β) components in the different treatments at four spatial scales: quadrats, within exclosures, within plots, within treatments.

3. At q = 0, α‐diversity was always significantly lower, and β‐diversity significantly higher, than predicted by the randomised null model. Diversity partitioning at q = 2 showed a similar trend at the quadrat scale. At the exclosure scale, partitioning exhibited a similar trend during the first 2 years of secondary succession but did not deviate from the null model in the third year, as observed in protected vegetation in all years.

4. At q = 0, diversity decreased across all treatments in the short term. At q = 2, diversity was initially higher in grazed plots than in protected vegetation; α and β components both decreased during secondary succession, to the levels observed in the protected vegetation.

5. Synthesis and applications.. Lower dominance in grazed vegetation indicates that grazing affects competitive exclusion at the local, small scale and accentuates natural heterogeneity (e.g. patchiness of soil resources, presence of rocks in the landscape) at a larger scale. The results of this study emphasise the importance of grazing as a management tool for maintaining plant diversity at multiple scales. This is a major concern worldwide, as the area covered by natural ecosystems continues to dwindle, necessitating management of grasslands for multiple functions such as animal production, resource protection and wildlife enhancement.
Theatrical entertainments and kind words: nursing the insane in Western North Carolina, 1882–1907
Authors: STREETER C.
Published: June 2011

Accessible summary

At the end of the 19th century, US asylum superintendents viewed psychiatric nursing education among the most important needs at state and private hospitals. Well‐trained nurses were critical to successful implementation of moral therapy, the predominant therapeutic approach of the time.

Dr Patrick Murphy, superintendent of the State Hospital at Morganton (a rural western North Carolina town), began a nurse training school in 1895. The school trained a corps of nurses who were the first in their families to enter a work in health care, nearly all of their fathers were farmers and their mothers kept house.

The nurses at the State Hospital at Morganton played a key role in shaping the public image about asylum care. They not only treated patients at the hospital, but also worked as private nurses in the community and as ‘nerve’ nurses at boarding homes and resort towns nearby. These nurses demonstrated laudable achievements, and were hired by other asylum superintendents who were struggling to implement nursing reforms in their hospitals.

This paper argues that at the turn of the 19th century, nurses at the State Hospital in Morganton, North Carolina (now called Broughton Hospital) played critical roles in successfully implementing the best‐known therapeutic methods of the time. They were also instrumental in developing the hospital's visibility and acceptance in rural western North Carolina. When the Hospital established its first nurse training school in 1895, this corps of first‐generation western North Carolinians practising institutional nursing was highly esteemed in their field. Their skills not only served the community outside of the Hospital's walls, but were also sought out by other private and state asylums.
New natural host plants of ‘Candidatus Phytoplasma pini’ in Poland and the Czech Republic
Authors: Kamińska M. , Berniak H. , Obdrzalek J.
Published: June 2011

The presence of phytoplasmas in seven coniferous plant species (Abies procera, Pinus banksiana, P. mugo, P. nigra, P. sylvestris, P. tabuliformis and Tsuga canadensis) was demonstrated using nested PCR with the primer pairs P1/P7 followed by R16F2n/R16R2. The phytoplasmas were detected in pine trees with witches’ broom symptoms growing in natural forest ecosystems and also in plants propagated from witches’ brooms. Identification of phytoplasmas was done using restriction fragment length polymorphism analysis (RFLP) of the 16S rDNA gene fragment with AluI, MseI and RsaI endonucleases. All samples showed RFLP patterns similar to the theoretical pattern of ‘Candidatus Phytoplasma pini’, based on the sequence of the reference isolate Pin127S. Nested PCR‐amplified products, obtained with primers R16F2n/R16R2, were sequenced. Comparison of the 16S rDNAs obtained revealed high (99·8–100%) nucleotide sequence identity between the phytoplasma isolates. The isolates were also closely related to four other phytoplasma isolates found in pine trees previously. Based on the results of RFLP and sequence analyses, the phytoplasma isolates tested were classified as members of the ‘Candidatus Phytoplasma pini’, group 16SrXXI.
Expression of Interleukin‐22 in Sjögren’s Syndrome: Significant Correlation with Disease Parameters
Authors: Lavoie T. N. , Stewart C. M. , Berg K. M. , Li Y. , Nguyen C. Q.
Published: June 2011

Sjögren’s syndrome (SS) is an autoimmune disease targeting the exocrine glands resulting in xerostomia/keratoconjunctivitis sicca. Presently, we examined the levels and clinical correlations of IL‐22 in SS. Patients with SS together with normal controls were randomly selected. IL‐22 was detected at significantly higher levels in sera of patients with SS. The levels of IL‐22 present in sera showed statistically significant direct correlations with hyposalivation, anti‐SSB, anti‐SSA/SSB combined, hypergammaglobulinemia and rheumatoid factor. IL‐22 showed a direct correlation with major clinical parameters. The data suggest that IL‐22 plays a critical role in the development of SS, and further study is needed to examine its function in human SS.
Pattern of Pre‐existing IgG Subclass Responses to a Panel of Asexual Stage Malaria Antigens Reported During the Lengthy Dry Season in Daraweesh, Sudan
Authors: Nasr A. , Iriemenam N. C. , Troye‐Blomberg M. , Arnot D. , Theander T. G. , Berzins K. , Giha H. A. , ElGhazali G.
Published: June 2011

The anti‐malarial IgG immune response during the lengthy and dry season in areas of low malaria transmission as in Eastern Sudan is largely unknown. In this study, ELISA was used for the measurement of pre‐existing total IgG and IgG subclasses to a panel of malaria antigens, MSP2‐3D7, MSP2‐FC27, AMA‐1 and Pf332‐C231. The results showed that the antibody responses were predominantly age dependent, antigen specific, and their lifespan was at least 5–6 month long. Generally, the IgG3 was most abundant IgG subclass, and the most recognized antigen was Pf332‐C231. Furthermore, the correlation between the levels of IgG subclasses was strongest between IgG1 and IgG3, which were more predictive to the total IgG levels. Finally, the response pattern of each of the IgG subclasses to the different test antigens that were spanning the dry season and the correlation between these responses were described in details for the first time.
Novel IFNγ ELISPOT Assay for Detection of Functional Carcinoembryonic Antigen‐Specific Chimeric Antigen Receptor‐Redirected T Cells
Authors: Elkord E. , Burt D. J.
Published: June 2011

We here describe the development of a novel ELISPOT assay for the detection and enumeration of IFNγ‐secreting functional chimeric antigen receptor (CAR)‐redirected T cells against carcinoembryonic antigen (CEA). This method is valuable for clinical trials to monitor the presence of functional CEA‐specific T cells transduced with a CAR. The same principle should be applicable for the detection of functional CAR‐redirected T cells against any other tumour‐associated antigens by immobilizing a particular biotinylated antigen to streptavidin‐coated beads.
Lysosomal Membrane Permeabilization Induces Cell Death in Human Mast Cells
Authors: Melo F. R. , Lundequist A. , Calounova G. , Wernersson S. , Pejler G.
Published: June 2011

Mast cells (MC) have pathogenic roles in numerous disorders, and strategies that stabilize MC or induce MC apoptosis are therefore emerging as possible therapeutic regimens. A typical feature of MC is their high content of secretory lysosomes (granules), containing numerous components such as biogenic amines, cytokines, serglycin proteoglycan and proteases. Damage to the secretory lysosomes will thus lead to leakage of these compounds, including the proteases, into the cytosol, and this could potentially trigger apoptosis. Here, we evaluated whether MC are sensitive to cell death induced by secretory lysosome destabilization, induced by the lysosomotropic agent Leu‐Leu‐OMe (LLME). Human MC were sensitive to LLME‐induced cell death. In contrast, fibroblasts and HEK‐293 cells were largely resistant. As judged by Annexin V/propidium iodide staining, LLME caused apoptotic cell death, and this was supported by induction of caspase‐3‐like activity, detection of activated caspase‐3 by immunoblot analysis and reduced cell death in the presence of a caspase inhibitor. In support of a role for serglycin in regulating LLME‐induced cell death, the survival rate of various cell types correlated negatively with the level of serglycin expression. In summary, this study introduces the concept of using lysosomotropic agents to induce cell death of human MC.
Type‐A response regulators are required for proper root apical meristem function through post‐transcriptional regulation of PIN auxin efflux carriers
Authors: Zhang Wenjing , To Jennifer P. C. , Cheng Chia‐Yi , Eric Schaller G. , Kieber Joseph J.
Published: June 2011

The phytohormones cytokinin and auxin regulate a diverse array of plant processes, often acting together to modulate growth and development. Although much has been learned with regard to how each of these hormones act individually, we are just beginning to understand how these signals interact to achieve an integrated response. Previous studies indicated that exogenous cytokinin has an effect on the transcription of several PIN efflux carriers. Here we show that disruption of type‐A Arabidopsis response regulators (ARRs), which are negative regulators of cytokinin signalling, alters the levels of PIN proteins and results in increased sensitivity to N‐1‐naphthylphthalamic acid, an inhibitor of polar auxin transport. Disruption of eight of the 10 type‐A ARR genes affects root development by altering the size of the apical meristem. Furthermore, we show that the effect of cytokinin on PIN abundance occurs primarily at the post‐transcriptional level. Alterations of PIN levels in the type‐A ARR mutants result in changes in the distribution of auxin in root tips as measured by a DR5::GFP reporter, and an altered pattern of cell division and differentiation in the stem cell niche in the root apical meristem. Together, these data indicate that cytokinin, acting through the type‐A ARRs, alters the level of several PIN efflux carriers, and thus regulates the distribution of auxin within the root tip.
ATG2, an autophagy‐related protein, negatively affects powdery mildew resistance and mildew‐induced cell death in Arabidopsis
Authors: Wang Yiping , Nishimura Marc T. , Zhao Ting , Tang Dingzhong
Published: June 2011

The molecular interactions between Arabidopsis and the pathogenic powdery mildew Golovinomyces cichoracearum were studied by characterizing a disease‐resistant Arabidopsis mutant atg2‐2. The atg2‐2 mutant showed enhanced resistance to powdery mildew and dramatic mildew‐induced cell death as well as early senescence phenotypes in the absence of pathogens. Defense‐related genes were constitutively activated in atg2‐2. In atg2‐2 mutants, spontaneous cell death, early senescence and disease resistance required the salicylic acid (SA) pathway, but interestingly, mildew‐induced cell death was not fully suppressed by inactivation of SA signaling. Thus, cell death could be uncoupled from disease resistance, suggesting that cell death is not sufficient for resistance to powdery mildew. ATG2 encodes autophagy‐related 2, a protein known to be involved in the early steps of autophagosome biogenesis. The atg2‐2 mutant exhibited typical autophagy defects in autophagosome formation. Furthermore, mutations in several other ATG genes, including ATG5, ATG7 and ATG10, exhibited similar powdery mildew resistance and mildew‐induced cell death phenotypes. Taken together, our findings provide insights into the role of autophagy in cell death and disease resistance, and may indicate general links between autophagy, senescence, programmed cell death and defense responses in plants.
Exome sequencing of two patients in a family with atypical X‐linked leukodystrophy
Authors: Tsurusaki Y , Okamoto N , Suzuki Y , Doi H , Saitsu H , Miyake N , Matsumoto N
Published: June 2011

Tsurusaki Y, Okamoto N, Suzuki Y, Doi H, Saitsu H, Miyake N, Matsumoto N. Exome sequencing of two patients in a family with atypical X‐linked leukodystrophy.

We encountered a family with two boys similarly showing brain atrophy with reduced white matter, hypoplasia of the brain stem and corpus callosum, spastic paralysis, and severe growth and mental retardation without speaking a word. The phenotype of these patients was not compatible with any known type of syndromic leukodystrophy. Presuming an X‐linked disorder, we performed next‐generation sequencing (NGS) of the transcripts of the entire X chromosome. A single lane of exome NGS in each patient was sufficient. Six potential mutations were found in both affected boys. Two missense mutations, including c.92T>C (p.V31A) in L1CAM, were potentially pathogenic, but this remained inconclusive. The other four could be excluded. Because the patients did not show adducted thumbs or hydrocephalus, the L1CAM change in this family can be interpreted as different scenarios. Personal genome analysis using NGS is certainly powerful, but interpretation of the data can be a substantial challenge requiring a lot of tasks.
A nasally applied cellulose powder in seasonal allergic rhinitis (SAR) in children and adolescents; reduction of symptoms and relation to pollen load
Authors: Åberg Nils , Dahl Å. , Benson M.
Published: June 2011

Åberg N, Dahl Å, Benson M. A nasally applied cellulose powder in seasonal allergic rhinitis (SAR) in children and adolescents; reduction of symptoms and relation to pollen load. Pediatr Allergy Immunol 2011; 22: 594–599.

Background:  A nasally applied cellulose powder is increasingly used in many countries as a remedy for allergic rhinitis. The absence of side effects makes the treatment particularly attractive in children. The efficacy in pollen allergic children, however, is not studied, nor is the relation to various pollen exposures.

Methods:  During the birch pollen season in 2009, a double blind, placebo‐controlled study was conducted in 53 subjects, aged 8–18 yr, with allergic rhinitis attributed to birch pollen. All children were on daily oral antihistamine. Reminders and reporting of symptom scores were made by SMS on mobile phones. Pollen was collected in a volumetric trap from which figures of pollen concentrations from 1979 to 2009 were available.

Results:  There was a significant reduction in total symptom scores from the nose (Placebo 7.29, Active 6.07, p = 0.033) and specifically for running nose (Placebo 2.56, Active 2.03, p = 0.017). All symptoms from the nose, eyes and lower airways were lower in the active group but reached significance only as earlier. The best effect was seen after days with low or moderate pollen counts (≤100/m3), the predominating pollen load over 31 yr in the area. No clinically significant adverse effects were seen.

Conclusions:  The product reduces symptoms of SAR in children and adolescents. Original data on pollen concentrations over 31 yr are presented with levels mainly in the low range favouring the observed efficacy profile. SMS communication on mobile phone for reminders and recording symptom scores was an excellent logistics tool.
Educational case series: Mechanisms of drug allergy
Authors: Caubet Jean‐Christoph , Pichler Werner J. , Eigenmann Philippe A.
Published: June 2011

To cite this article: Caubet J‐C, Pichler WJ, Eigenmann PA. Educational case series: Mechanisms of drug allergy. Pediatr Allergy Immunol 2011; 22: 559–567.

Once administered, a drug can activate the immune system by various mechanisms and lead to a large range of clinical manifestations closely related to the type of immune reaction elicited. Administration of the drug can classically result in an immunoglobulin E (IgE)‐type sensitization, but can also result in more complex activation of the immune system potentially resulting in severe syndromes, such as the drug‐induced hypersensitivity syndrome (DIHS). Although there has been a major increase in our knowledge over the last years, the exact mechanisms of drug allergy are not well understood for most clinical manifestations. A complex interaction between individual characteristics, environmental factors, and the drug itself is usually responsible for adverse reactions to drugs. In this educational review series, we described three cases of drug allergy: first, a child with a typical IgE‐mediated drug allergy, second, a child with a non‐immediate reaction to penicillin, and in the third patient, we will discuss the drug‐induced hypersensitivity syndrome, which is rare but potentially fatal. These cases are correlated to the immune mechanism potentially involved.
Cloning and characterization of barley caryopsis FCA
Authors: Kumar Santosh , Jiang Shiling , Jami Sravan K. , Hill Robert D.
Published: June 2011

The RNA binding protein, flowering control locus A, (FCA) regulates flowering in rice and Arabidopsis. FCA interacts with FY to auto‐regulate its own transcripts as well as to control flowering by downregulating flowering locus C (FLC). We report the cloning and characterization of the gamma (γ) isoform of FCA from barley (Hordeum vulgare cv. McLeod). The deduced protein contained two RNA recognition motifs (RRMs), a glycine‐rich region at the N‐terminal end, a polyglutamine region immediately downstream of a WW domain. Barley FCA had greater protein sequence homology to wheat and rice FCA than to its Arabidopsis homolog. In developing barley embryos, FCA transcripts could be detected from 2 days after pollination (DAP) up to 40 DAP. FCA transcript levels in mature barley embryo were more abundant in non‐germinated than in germinated seeds, with the levels declining as germination progressed. ABA inhibition of germination inhibited the decline of barley embryo FCA. Transient co‐expression of FCA or a truncated FCA (lacking RRM) with maize VP1 promoter or wheat Em gene promoter in barley aleurone protoplasts led to increased VP1 and Em gene promoter activity. Barley FCA or truncated FCA localized in the nucleus suggested its role in gene regulation.
Variations of DNA methylation in Eucalyptus urophylla×Eucalyptus grandis shoot tips and apical meristems of different physiological ages
Authors: Mankessi François , Saya Aubin R. , Favreau Bénédicte , Doulbeau Sylvie , Conéjéro Geneviève , Lartaud Marc , Verdeil Jean‐Luc , Monteuuis Olivier
Published: June 2011

Global DNA methylation was assessed by high‐performance liquid chromatography (HPLC) for the first time in Eucalyptus urophylla×Eucalyptus grandis shoot tips comparing three outdoor and one in vitro sources of related genotypes differing in their physiological age. The DNA methylation levels found were consistent with those reported for other Angiosperms using the same HPLC technology. Notwithstanding noticeable time‐related fluctuations within each source of plant material, methylation rate was overall higher for the mature clone (13.7%) than for the rejuvenated line of the same clone (12.6%) and for the juvenile offspring seedlings (11.8%). The in vitro microshoots of the mature clone were less methylated (11.3%) than the other outdoor origins, but the difference with the juvenile seedlings was not significant. Immunofluorescence investigations on shoot apices established that the mature source could be distinguished from the rejuvenated and juvenile origins by a higher density of cells with methylated nuclei in leaf primordia. Shoot apical meristems (SAMs) from the mature clone also showed a greater proportion and more methylated cells than SAMs from the rejuvenated and juvenile origins. The nuclei of these latter were characterized by fewer and more dispersed labeled spots than for the mature source. Our findings establish that physiological ageing induced quantitative and qualitative variations of DNA methylation at shoot tip, SAM and even cellular levels. Overall this DNA methylation increased with maturation and conversely decreased with rejuvenation to reach the lower scores and to show the immunolabeling patterns that characterized juvenile material nuclei.
Slowly developing drought stress increases photosynthetic acclimation of Catharanthus roseus
Authors: Kim Jongyun , van Iersel Marc W.
Published: June 2011

Our understanding of plant responses to drought has improved over the decades. However, the importance of the rate of drought imposition on the response is still poorly understood. To test the importance of the rate at which drought stress develops, whole‐plant photosynthesis (Pnet), respiration (Rdark), daily carbon gain (DCG), daily evapotranspiration (DET) and water use efficiency (WUE) of vinca (Catharanthus roseus), subjected to different drought imposition rates, were investigated. We controlled the rate at which the substrate dried out with an automated irrigation system that allowed pot weight to decrease gradually throughout the drying period. Fast, intermediate and slow drying treatments reached their final pot weight [500 g, substrate water content (θ) ≈ 0.10 m3 m−3] after 3.1, 6.6 and 10 days, respectively. Although all drying treatments decreased Pnet and Rdark, slow drying reduced Pnet and Rdark less than fast drying. At a θ < 0.10 m3 m−3, DCG and DET in the slow drying treatment were reduced by ≈50%, whereas DCG and DET in the fast drying treatment were reduced by 85 and 70% at a θ of 0.16 m3 m−3. Plants exposed to slow drought imposition maintained a high WUE, even at θ < 0.10 m3 m−3. Overall, physiological responses to low θ were less severe in plants subjected to slow drying as compared with fast drying, even though the final θ was lower for plants exposed to slow drying. This suggests that the rate at which drought stress develops has important implications for the level of acclimation that occurs.
Prediction of flowering time in Brassica oleracea using a quantitative trait loci‐based phenology model
Authors: Uptmoor R. , Li J. , Schrag T. , Stützel H.
Published: June 2011

Uniformly developing plants with a predictable time to harvest or flowering under unfavourable climate conditions are a major breeding goal in crop species. The main flowering regulators and their response to environmental signals have been identified in Arabidopsis thaliana and homologues of flowering genes have been mapped in many crop species. However, it remains unclear which genes determine within and across genotype flowering time variability in Brassica oleracea and how genetic flowering time regulation is influenced by environmental factors. The goal of this study is model‐based prediction of flowering time in a B. oleracea DH‐line population using genotype‐specific and quantitative trait loci (QTL) model input parameters. A QTL‐based phenology model accounting for genotypic differences in temperature responses during vernalisation and non‐temperature‐sensitive durations from floral transition to flowering was evaluated in two field trials. The model was parameterised using original genotype‐specific model input parameters and QTL effects. The genotype‐specific model parameterisation showed accurate predictability of flowering time if floral induction was promoted by low temperature (R2 = 0.81); unfavourably high temperatures reduced predictability (R2 = 0.65). Replacing original model input parameters by QTL effects reduced the capability of the model to describe across‐genotype variability (R2 = 0.59 and 0.50). Flowering time was highly correlated with a model parameter accounting for vernalisation effects. Within‐genotype variability was significantly correlated with the same parameter if temperature during the inductive phase was high. We conclude that flowering time variability across genotypes was largely due to differences in vernalisation response, although it has been shown elsewhere that the candidate FLOWERING LOCUS C (FLC) did not co‐segregate with flowering time in the same population. FLC independent vernalisation pathways have been described for several species, but not yet for B. oleracea.