bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of Auxins and Cytokinins in the Mutualistic Interaction Between Arabidopsis and Piriformospora indica
Authors: Vadassery Jyothilakshmi , Ritter Claudia , Venus Yvonne , Camehl Iris , Varma Ajit , Shahollari Bationa , Novák Ondrej , Strnad Miroslav , Ludwig-Müller Jutta , Oelmüller Ralf
Published: September 2008

Arabidopsis growth and reproduction are stimulated by the endophytic fungus Piriformospora indica. The fungus produces low amounts of auxins, but the auxin levels and the expression of auxin-regulated genes are not altered in colonized roots. Also, mutants with reduced auxin levels (ilr1-1, nit1-3, tfl2, cyp79 b2b3) respond to P. indica. However, the fungus rescues the dwarf phenotype of the auxin overproducer sur1-1 by converting free auxin into conjugates, which also results in the downregulation of the auxin-induced IAA6 and the upregulation of the P. indica–induced LRR1 gene. The fungus produces relatively high levels of cytokinins, and the cytokinin levels are higher in colonized roots compared with the uncolonized controls. trans-Zeatin cytokinin biosynthesis and the CRE1/AHK2 receptor combination are crucial for P. indica–mediated growth stimulation, while mutants lacking cis-zeatin, impaired in other cytokinin receptor combinations, or containing reduced cytokinin levels respond to the fungus. Since root colonization is not affected in the cytokinin mutants, we propose that cytokinins are required for P. indica–induced growth promotion. Finally, a comparative analysis of the phytohormone mutants allows the conclusion that the response to P. indica is independent of the architecture and size of the roots.
Effect of Southern Root-knot Nematode (Meloidogyne incognita race 3) on Corn Yields in Alabama
Authors: Bowen K. L. , Hagan A. K. , Campbell H. L. , Nightengale S.
Published: September 2008

In Alabama and other southeastern states, corn is frequently planted in rotation with cotton and peanut in order to minimize soil-borne pest problems. Even in areas where peanut is not grown, corn is increasingly being planted in rotation with cotton. However, one root-knot nematode, Meloidogyne incognita race 3, causes damage to both cotton and corn. In this study, we determined levels of corn grain loss when southern root-knot nematodes are present in soil. Losses were 3.8 to 11.4% based on preceding years' counts and 2.2 to 7.0% with current years' counts for every 100 2nd-stage juvenile root-knot nematodes in 100 cm ≥ of soil. Knowledge of the percent loss in corn grain due to southern rootknot nematode populations can provide additional guidance for use of risk thresholds when growers are making management decisions.

Accepted for publication 21 June 2008. Published 10 September 2008.
Plant-Pathogenic Bacteria as Biological Weapons – Real Threats?
Authors: Young J. M. , Allen C. , Coutinho T. , Denny T. , Elphinstone J. , Fegan M. , Gillings M. , Gottwald T. R. , Graham J. H. , Iacobellis N. S. , Janse J. D. , Jacques M.-A. , Lopez M. M. , Morris C. E. , Parkinson N. , Prior P. , Pruvost O. , Neto J. Rodrigues , Scortichini M. , Takikawa Y. , Upper C. D.
Published: September 2008

At present, much attention is being given to the potential of plant pathogens, including plant-pathogenic bacteria, as biological weapons/bioterror weapons. These two terms are sometimes used interchangeably and there is need for care in their application. It has been claimed that clandestine introduction of certain plant-pathogenic bacteria could cause such crop losses as to impact so significantly on a national economy and thus constitute a threat to national security. As a separate outcome, it is suggested that they could cause serious public alarm, perhaps constituting a source of terror. Legislation is now in place to regulate selected plant-pathogenic bacteria as potential weapons. However, we consider it highly doubtful that any plant-pathogenic bacterium has the requisite capabilities to justify such a classification. Even if they were so capable, the differentiation of pathogens into a special category with regulations that are even more restrictive than those currently applied in quarantine legislation of most jurisdictions offers no obvious benefit. Moreover, we believe that such regulations are disadvantageous insofar as they limit research on precisely those pathogens most in need of study. Whereas some human and animal pathogens may have potential as biological or bioterror weapons, we conclude that it is unlikely that any plant-pathogenic bacterium realistically falls into this category.
Occurrence and Characterization of a Phytophthora sp. Pathogenic to Asparagus (Asparagus officinalis) in Michigan
Authors: Saude C. , Hurtado-Gonzales O. P. , Lamour K. H. , Hausbeck M. K.
Published: September 2008

A homothallic Phytophthora sp. was recovered from asparagus (Asparagus officinalis) spears, storage roots, crowns, and stems in northwest and central Michigan in 2004 and 2005. Isolates (n = 131) produced ovoid, nonpapillate, noncaducous sporangia 45 μm long × 26 μm wide and amphigynous oospores of 25 to 30 μm diameter. Mycelial growth was optimum at 25°C with no growth at 5 and 30°C. All isolates were sensitive to 100 ppm mefenoxam. Pathogenicity studies confirmed the ability of the isolates to infect asparagus as well as cucurbits. Amplified fragment length polymorphism analysis of 99 isolates revealed identical fingerprints, with 12 clearly resolved fragments present and no clearly resolved polymorphic fragments, suggesting a single clonal lineage. The internal transcribed spacer regions of representative isolates were homologous with a Phytophthora sp. isolated from diseased asparagus in France and a Phytophthora sp. from agave in Australia. Phylogenetic analysis supports the conclusion that the Phytophthora sp. isolated from asparagus in Michigan is a distinct species, and has been named Phytophthora asparagi.
Proteomic Analysis Reveals Suppression of Bark Chitinases and Proteinase Inhibitors in Citrus Plants Affected by the Citrus Sudden Death Disease
Authors: Cantú M. D. , Mariano A. G. , Palma M. S. , Carrilho E. , Wulff N. A.
Published: September 2008

Citrus sudden death (CSD) is a disease of unknown etiology that greatly affects sweet oranges grafted on Rangpur lime rootstock, the most important rootstock in Brazilian citriculture. We performed a proteomic analysis to generate information related to this plant pathogen interaction. Protein profiles from healthy, CSD-affected and CSD-tolerant stem barks, were generated using two-dimensional gel electrophoresis. The protein spots were well distributed over a pI range of 3.26 to 9.97 and a molecular weight (MW) range from 7.1 to 120 kDa. The patterns of expressed proteins on 2-DE gels made it possible to distinguish healthy barks from CSD-affected barks. Protein spots with MW around 30 kDa and pI values ranging from 4.5 to 5.2 were down-regulated in the CSD-affected root-stock bark. This set of protein spots was identified as chitinases. Another set of proteins, ranging in pI from 6.1 to 9.6 with an MW of about 20 kDa, were also suppressed in CSD-affected rootstock bark; these were identified as miraculin-like proteins, potential trypsin inhibitors. Down-regulation of chitinases and proteinase inhibitors in CSD-affected plants is relevant since chitinases are well-known pathogenesis-related protein, and their activity against plant pathogens is largely accepted.
Nitrite Reductase Gene Upregulated During Conidiation Is Involved in Macroconidium Formation in Fusarium oxysporum
Authors: Iida Y. , Kurata T. , Harimoto Y. , Tsuge T.
Published: September 2008

Fusarium oxysporum produces three kinds of asexual spores, microconidia, macroconidia, and chlamydospores. We previously found that the transcript level of the nitrite reductase gene of F. oxysporum, named FoNIIA, was markedly upregulated during conidiation compared with during vegetative growth. FoNIIA was also found to be positively regulated by Ren1 that is a transcription regulator controlling development of microconidia and macroconidia. In this study, we analyzed the function of FoNIIA in conidiation of F. oxysporum. Conidiation cultures showed markedly higher level of accumulation of FoNiiA protein as well as FoNIIA mRNA than vegetative growth cultures. FoNIIA protein was significantly decreased in cultures of the REN1 disruption mutant compared with that of the wild type. These results confirmed that FoNIIA expression is upregulated during conidiation and is positively regulated by REN1. The FoNIIA disruption mutants produced microconidia, macroconidia, and chlamydospores, which were morphologically indistinguishable from those of the wild type. The mutants, however, produced significantly fewer macroconidia than the wild type, although the wild type and mutant strains produced similar numbers of microconidia and chlamydospores. These results demonstrate that nitrite reductase is involved in quantitative control of macroconidium formation as well as nitrate utilization in F. oxysporum.
Assessment of Common Scab-Inducing Pathogen Effects on Potato Underground Organs Via Computed Tomography Scanning
Authors: Han L. , Dutilleul P. , Prasher S. O. , Beaulieu C. , Smith D. L.
Published: September 2008

Common scab caused by Streptomyces scabies is a major bacterial disease of potato (Solanum tuberosum). Its best known symptom is superficial lesions on the surface of progeny potato tubers, observed at harvesting. In this study, effects of S. scabies on space occupancy by underground organs and on structural complexity of root systems are investigated during growth via computed tomography (CT) scanning. Two groups of potato plants were grown in a greenhouse in middle-sized plastic pots. Using a high-resolution X-ray CT scanner formerly used for medical applications, their underground organs and surrounding medium (sieved and autoclaved homogeneous sand) were submitted to CT scanning 4, 6, and 8 weeks after planting. For one group, sand was inoculated with the common scab-inducing pathogen (S. scabies EF-35) at potting. Space occupancy by underground organs was estimated via curve fitting applied to histograms of CT scan data, while three-dimensional skeletal images were used for fractal analysis. Root systems of diseased plants were found to be less complex than those of healthy plants 4 weeks after planting, and the relative growth rates derived from space occupancy measures were of different sign between the two groups from week 4 to week 8.
Antibiosis and Acidification byPantoea agglomeransStrain E325 May Contribute to Suppression ofErwinia amylovora
Authors: Pusey P. L. , Stockwell V. O. , Rudell D. R.
Published: September 2008

Pantoea agglomerans strain E325, a commercially available antagonist for fire blight of apple and pear, was originally selected through screening based on suppression of Erwinia amylovora on flower stigmas, but specific mechanisms of antagonism were unknown. Bacterial modification of pH was evaluated as a possible mechanism by analyzing stigma exudates extracted from ‘Gala’ apple stigmas. The pH values for field samples were only slightly lower than controls, but indicated a range (pH 5 to 6) conducive for antibiotic activity according to subsequent assays. Under low-phosphate and low-pH conditions, an antibacterial product of E325 with high specificity to E. amylovora was effective at low concentrations. A minimum of 20 to 40 ng of a ninhydrin-reactive compound purified using RP-HPLC caused visible inhibition in assays. Activity was heat stable and unaffected by amino acids, iron, or enzymes known to affect antibiotics of P. agglomerans. Antibiosis was diminished, however, under basic conditions, and with increasing phosphate concentrations at pH 6 and 7. Inhibition was not observed in media containing phosphate concentrations commonly used in antibiosis assays. We propose that E325 suppresses the fire blight pathogen not only by competing for nutrients on the stigma, but by producing an antibiotic specific to E. amylovora. Further work is necessary to substantiate that the compound is produced and active on flower stigmas.
Functional Specialization of the TRANSPARENT TESTA GLABRA1 Network Allows Differential Hormonal Control of Laminal and Marginal Trichome Initiation in Arabidopsis Rosette Leaves
Authors: Maes Lies , Inzé Dirk , Goossens Alain
Published: September 2008

Trichome initiation in Arabidopsis (Arabidopsis thaliana) is controlled by the TRANSPARENT TESTA GLABRA1 (TTG1) network that consists of R2R3- and R1-type MYB-related transcription factors, basic helix-loop-helix (bHLH) proteins, and the WD40 protein TTG1. An experimental method was designed to investigate the molecular mechanisms by which jasmonates, cytokinins, and gibberellins modulate Arabidopsis leaf trichome formation. All three phytohormones provoked a seemingly common effect on cell patterning by promoting trichome initiation but caused strikingly distinct effects on cell and trichome maturation. The phytohormonal control was mediated by transcriptional regulation of the established TTG1 complex and depended on the R2R3-MYB factor GLABRA1. However, unsuspected degrees of functional specialization of the bHLH factors and a resultant differential molecular regulation of trichome initiation on leaf lamina and leaf margins were revealed. Trichome formation on leaf lamina relied entirely on GLABRA3 and ENHANCER OF GLABRA3. Conversely, TRANSPARENT TESTA8 (TT8) was particularly important for marginal trichome development. This hitherto unknown role for TT8 in trichome formation further underscored the functional redundancy between the three TTG1-dependent bHLH proteins.
Complex I Dysfunction Redirects Cellular and Mitochondrial Metabolism in Arabidopsis
Authors: Garmier Marie , Carroll Adam J. , Delannoy Etienne , Vallet Corinne , Day David A. , Small Ian D. , Millar A. Harvey
Published: September 2008

Mitochondrial complex I is a major avenue for reduced NAD oxidation linked to oxidative phosphorylation in plants. However, the plant enzyme has structural and functional features that set it apart from its counterparts in other organisms, raising questions about the physiological significance of this complex in plants. We have developed an experimental model in which rotenone, a classic complex I inhibitor, has been applied to Arabidopsis (Arabidopsis thaliana) cell suspension cultures in order to dissect early metabolic adjustments involved in cell acclimation to mitochondrial dysfunction. Rotenone induced a transitory decrease in cellular respiration (0–4 h after treatment). Cell respiration then progressively recovered and reached a steady state at 10 to 12 h after treatment. Complex I inhibition by rotenone did not induce obvious oxidative stress or cell death but affected longer term cell growth. Integrated analyses of gene expression, the mitochondrial proteome, and changes in primary metabolism indicated that rotenone treatment caused changes in mitochondrial function via alterations in specific components. A physical disengagement of glycolytic activities associated with the mitochondrial outer membrane was observed, and the tricarboxylic acid cycle was altered. Amino acid and organic acid pools were also modified by rotenone treatment, with a marked early decrease of 2-oxoglutarate, aspartate, and glutamine pools. These data demonstrate that, in Arabidopsis cells, complex I inhibition by rotenone induces significant remodeling of metabolic pathways involving the mitochondria and other compartments and point to early metabolic changes in response to mitochondrial dysfunction.
Different Hormonal Regulation of Cellular Differentiation and Function in Nucellar Projection and Endosperm Transfer Cells: A Microdissection-Based Transcriptome Study of Young Barley Grains
Authors: Thiel Johannes , Weier Diana , Sreenivasulu Nese , Strickert Marc , Weichert Nicola , Melzer Michael , Czauderna Tobias , Wobus Ulrich , Weber Hans , Weschke Winfriede
Published: September 2008

Nucellar projection (NP) and endosperm transfer cells (ETC) are essential tissues in growing barley (Hordeum vulgare) grains, responsible for nutrient transfer from maternal to filial tissues, endosperm/embryo nutrition, and grain development. A laser microdissection pressure catapulting-based transcriptome analysis was established to study NP and ETC separately using a barley 12K macroarray. A major challenge was to isolate high-quality mRNA from preembedded, fixed tissue while maintaining tissue integrity. We show that probes generated from fixed and embedded tissue sections represent largely the transcriptome (>70%) of nonchemically treated and nonamplified references. In NP, the top-down gradient of cellular differentiation is reflected by the expression of C3HC4-type ubiquitin ligases and different histone genes, cell wall biosynthesis and expansin/extensin genes, as well as genes involved in programmed cell death-related proteolysis coupled to nitrogen remobilization, indicating distinct areas simultaneously undergoing mitosis, cell elongation, and disintegration. Activated gene expression related to gibberellin synthesis and function suggests a regulatory role for gibberellins in establishment of the differentiation gradient. Up-regulation of plasmalemma-intrinsic protein and tonoplast-intrinsic protein genes indicates involvement in nutrient transfer and/or unloading. In ETC, AP2/EREBP-like transcription factors and ethylene functions are transcriptionally activated, a response possibly coupled to activated defense mechanisms. Transcriptional activation of nucleotide sugar metabolism may be attributed to ascorbate synthesis and/or cell wall biosynthesis. These processes are potentially controlled by trehalose-6-P synthase/phosphatase, as suggested by expression of their respective genes. Up-regulation of amino acid permeases in ETC indicates important roles in active nutrient uptake from the apoplastic space into the endosperm.
Detection of QTL for growth rate in the blacklip abalone (Haliotis rubra Leach) using selective DNA pooling
Authors: Baranski M. , Rourke M. , Loughnan S. , Hayes B. , Austin C. , Robinson N.
Published: September 2008

The objective of this study was to identify QTL for growth rate in the blacklip abalone Haliotis rubra using selective DNA pooling. Three full‐sibling families of H. rubra derived from crosses of wild broodstock were used. DNA was extracted from the largest and smallest 10% of progeny and combined into two pools for each phenotypic tail. The DNA pools were typed with 139 microsatellites, and markers showing significant differences between the peak height ratios of alleles inherited from the parents were individually genotyped and analysed by interval mapping. A strong correlation (r = 0.94, P < 0.001) was found between the t‐values from the analysis of pools and the t‐values from the analysis of individual genotypes. Based on the interval mapping analysis, QTL were detected on nine linkage groups at a chromosome‐wide P < 0.01 and one linkage group at a chromosome‐wide P < 0.05. The study demonstrated that selective DNA pooling is efficient and effective as a first‐pass screen for the discovery of QTL in an aquaculture species.
Characterization of EST‐derived microsatellites for gene mapping and evolutionary genomics in turbot
Authors: Bouza C. , Hermida M. , Millán A. , Vilas R. , Vera M. , Fernández C. , Calaza M. , Pardo B. G. , Martínez P.
Published: September 2008

The detection of microsatellite sequences within expressed sequence tags (ESTs) connects potential markers with specific genes, generating type I markers. We have developed and mapped by linkage analysis a set of EST‐derived microsatellites in the turbot, Scophthalmus maximus. One hundred and ninety‐one microsatellites were identified from 9256 turbot ESTs. Primer design was possible with 98 microsatellites. After genotyping 25 wild turbot and the parents of two reference families for linkage analysis, 43 EST‐derived microsatellites were selected because they met technical and polymorphism criteria. A final set of 31 EST‐derived microsatellites could be mapped to 17 linkage groups of the turbot consensus map based on 242 anonymous microsatellites. Twenty‐four microsatellite‐containing ESTs were functionally annotated, confirming them as type I markers. Nineteen were mapped in the turbot consensus map. These EST‐derived microsatellites constitute useful tools for genome scanning of turbot populations, marker‐assisted selection programmes and comparative mapping.
Primary care treatment for child and adolescent neuropsychiatric conditions in remote rural Punjab, Pakistan – a cross‐sectional survey
Authors: Tareen A. , Mirza I. , Mujtaba M. , Chaudhry H. R. , Jenkins R.
Published: September 2008

Background  Pakistan is one of the most populous regions of the world. Previous work has demonstrated that there is reliance on traditional healthcare systems when seeking psychiatric care; however, there is a lack of information on help seeking for child and adolescent population. The aim of this study was to describe types of treatments and families’ perceptions of the effectiveness of those treatments for childhood neuropsychiatric disorders in remote rural Punjab.

Method  Cross‐sectional survey of consecutive attendees at an advertised mental health consultation day in a remote rural area.

Results  The effectiveness of these treatments, as rated by patients and their families, was variable, with highest reported effectiveness for general practitioner treatments. In families with a past history of care from a general practitioner, those who had epilepsy reported treatments to be more effective than those with mental retardation. Carers and users described consulting five different types of primary healthcare practitioners that used both physical and psychotherapeutic treatments.

Conclusions  There is considerable variation in treatments available for child and adolescent neuropsychiatric disorders in remote rural areas of Punjab, a large proportion of which are considered ineffective by the users and carers. This highlights the need to develop effective interventions for child and adolescent neuropsychiatric conditions that can be administered by primary health workers. Our data suggest that the need for this is greatest for mental retardation.
The relationship between proxy reported health‐related quality of life and parental distress: gender differences
Authors: Davis E. , Davies B. , Waters E. , Priest N.
Published: September 2008

Objective  Although primary caregiver proxy reports of health‐related quality of life (HRQOL) are often used for healthcare decision making when child self‐reports are unable to be collected (because of a variety of reasons such as child illness, disability or age), we have little understanding of the correlates of parent‐proxy reports. The aim of this study was to examine the relationship between parental depression and parent‐proxy reported QOL for primary caregivers (mothers and fathers), using a multidimensional HRQOL instrument. It was hypothesized that maternal depression would be negatively correlated with maternal reported HRQOL, but that paternal depression would not be correlated with paternal reported HRQOL.

Methods  Data were from parents of children aged 4–5 years (n = 4983) involved in the Longitudinal Study of Australian Children. A questionnaire assessing parental depression (Kessler‐6) and proxy reported HRQOL (Pediatric Quality of Life Inventory) was completed by the primary caregiver.

Results  For maternal primary caregivers, maternal depression was negatively correlated with all domains of maternal proxy reports of HRQOL (r = −0.24 to r = −0.36). For paternal primary caregivers, there was no relationship between paternal depression and paternal proxy reports of HRQOL. Multiple regression analyses demonstrated that maternal depression was a significant predictor of total HRQOL, accounting for 12% of the variance. For paternal mental health, depression did not predict parent‐proxy reported total HRQOL.

Conclusion  These results highlight the importance of assessing maternal mental health when measuring proxy reported QOL. Further research is needed in this area to examine the relationship between parental depression and proxy reported HRQOL (including both mothers and fathers, where possible), as well as child self‐reported HRQOL.
Phytoplankton distribution along trophic gradients within and among reservoirs in Catalonia (Spain)
Authors: CAPUTO LUCIANO , NASELLI‐FLORES LUIGI , ORDOÑEZ JAIME , ARMENGOL JOAN
Published: September 2008

1. Longitudinal gradients in the epilimnetic waters of stratified reservoirs provide a useful database to study changing environmental conditions. The spatial distribution, assemblage structure and specific adaptations of phytoplankton assemblages can be analysed along these gradients over short time scales.

2. Four reservoirs with a similar typology, located along an altitudinal gradient in the same eco‐region, were sampled along their longitudinal axes. In total, 19 sampling stations provided a trophic spectrum, ranging from oligo‐mesotrophy to hypertrophy, which was quantified by calculating the trophic state index of each sampling station in the four reservoirs.

3. Several patterns in phytoplankton assemblage structure were detected. Total chlorophyll‐a (Chl‐a), biovolume, abundance and the relative biomass contribution of the main algal groups (chlorophytes, cyanobacteria, cryptophytes and diatoms) were highly correlated with their location along the trophic gradient.

4. We also adopted the functional classification of Reynolds et al. (2002): this effectively summarized differences among phytoplankton assemblages under varying resource‐limiting combinations, especially nutrients and underwater light climate.

5. In terms of relationships with the trophic gradient, diatoms and cyanobacteria exhibited significant opposing trends in both their relative chlorophyll contribution to total Chl‐a and biovolume. Chlorophytes were more abundant at an intermediate position along the trophic spectrum.

6. The identified patterns are consistent with models of self‐organization of phytoplankton assemblages. In particular, light availability was a strong determinant of size and shape diversity, especially in hypertrophic conditions, where ‘R‐strategist’, needle shaped species, dominated the system. In contrast, under decreased availability of nutrients and higher light extinction coefficients (Kd), the system was co‐dominated by C‐ and S‐strategist species, having shapes with a higher surface/volume ratio.
Relationship between the friction coefficient and velocity of two cactus pear (Opuntia ficus india L.) cultivars on different surfaces
Authors: Kabas Onder , Ozmerzi Aziz
Published: September 2008

The static and dynamic coefficients of friction were determined for two cactus pear cultivars against four different structural materials, namely galvanised steel, rubber, plywood and paper for surface moving velocities of 5.35–26.72 mm s−1. Velocity affected the dynamic coefficient more than the static coefficient. Differences between the tested surfaces were most pronounced at a velocity of 18.63 mm s−1. The effects of velocity and friction surface varied inconsistently between the two cultivars.
Radical scavenging and antimicrobial activity of horsetail (Equisetum arvense L.) extracts
Authors: Čanadanović‐Brunet Jasna M. , Ćetković Gordana S. , Djilas Sonja M. , Tumbas Vesna T. , Savatović Sladjana S. , Mandić Anamarija I. , Markov Siniša L. , Cvetković Dragoljub D.
Published: September 2008

A reversed‐phase high performance liquid chromatography (RP‐HPLC) separation on C8 column and quantitative method were developed to analyse hydroxyl derivatives of benzoic and cinnamic acid and flavonoids in horsetail (Equisetum arvense L.) extracts. Total phenolic content of n‐butanol, ethyl acetate and water extracts, determined by the Folin‐Ciocalteu method, was 96.4, 26.4 and 15.4 mg g−1 of dry extracts, respectively. The antioxidative activity of horsetail extracts was tested by measuring their ability to scavenge stable 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and reactive hydroxyl radicals by electron spin resonance spectroscopy. The results demonstrated that the free radical scavenging activity (versus both DPPH and hydroxyl radicals) depended on the type and concentration of applied extracts; the highest DPPH (EC50 = 0.65 mg mL−1) and hydroxyl radical scavenging activities (EC50 = 0.74 mg mL−1) were obtained in the case of n‐butanol extract. The radical scavenging activity of extracts significantly correlated with total phenolic content. The antimicrobial tests showed that ethyl acetate and n‐butanol extracts inhibited the growth of tested bacteria.
Protein stabilisation of Chardonnay wine using trisacryl and bentonite: a comparative study
Authors: De Bruijn Johannes , Loyola Cristina , Flores Adan , Hevia Felicitas , Melín Pedro , Serra Ignacio
Published: September 2008

The stabilisation of a Chilean Chardonnay wine by SP‐Trisacryl‐M and bentonite was investigated, evaluating protein, polyphenol and polysaccharide adsorption, turbidity and wine quality. The wine could be stabilised by adding at least 0.3 kg m−3 of bentonite or 12 kg m−3 of trisacryl, removing 95% and 76% of the wine proteins, respectively. The protein adsorption data for bentonite and trisacryl were fitted using the Freundlich isotherm. The wine protein adsorption isotherm on trisacryl was unfavourable. Protein removal from Chardonnay by trisacryl in a packed column at continuous operation was about 50% during the first 70 bed volumes (BV) of treated wine and decreased progressively until the end of the treatment (100 BV). The adsorbents showed a higher selectivity for proteins than for polyphenols and polysaccharides. A sensorial panel could not detect statistically significant differences between the bentonite and trisacryl treatments of wine at P ≤ 0.05.
Physico‐chemical studies on a wide composition range of low‐moisture glucose–fructose mixtures: rates of crystallisation
Authors: Arvanitoyannis Ioannis S.
Published: September 2008

The crystallisation rates of low‐moisture (from 2% to 10% w/w) glucose–fructose mixtures were investigated at a variety of storage temperatures (from 0 to 60 °C). It was found that d‐fructose considerably retards the rate of crystallisation. High storage temperatures induced a decrease in the ‘threshold’ moisture content, which is necessary for the initial nucleation and further development of the glucose and/or fructose crystals. This knowledge of crystallisation rates can be exploited in terms of storage of confectionery products containing d‐glucose, d‐fructose or both of them, and honey.
Detecting vegetable oil adulteration in hazelnut paste (Corylus avellana L.)
Authors: Bonvehi Josep Serra , Coll Francesc Ventura
Published: September 2008

The possibilities of detecting hazelnut paste adulterated with refined and non‐refined vegetable oils have been studied. Research was focussed mainly on peanut, high oleic‐acid sunflower, corn and soybean oils which have a similar composition to hazelnut oil. The analytical procedures to detect fatty acid (FA), triacylglycerol (TAG) and tocopherol profiles as indicators of adulteration were determined. The better indicators experimentally determined were seven FA (palmitic, stearic, linoleic, linolenic, arachidonic, behenic and lignoceric acids) and different TAG with three unsaturated FA (the code letters used for FA are: P = C16:0; S = C18:0; O = C18:1; L = C18:2;; Ln = C18:3) (LLLn, LLL and OOO), two unsaturated FA (POL, PLL and SOO), and one unsaturated FA (PPL). As expected, when refined vegetable oils were added to hazelnut paste, the increment of stigmasta‐3,5‐diene allowed detection at levels of 2% oil added. Limits of detection were measured using standard and adulterated hazelnut with different amounts of non‐refined vegetable oils added (5%, 10%, 20% and 30%). The distribution of tocopherols and tocotrienols is highly useful, except in the case of added sunflower oil. The differences between the experimental and theoretical values of the TAG with equivalent carbon number (ΔECN) of 42 does not improve the detection limit of hazelnut paste adulterated with peanut or sunflower oils. Similarly, tocopherols usually added to refined vegetable oils as an antioxidant were also determined.
Compositional variation in β‐carotene content, carbohydrate and antioxidant enzymes in selected banana cultivars
Authors: Arora Ajay , Choudhary Divya , Agarwal Gaurav , Singh Virender Pal
Published: September 2008

Banana is one of the most important tropical fruits, and India ranks first in its world production. Banana cultivars rich in provitamin A carotenoids may offer a potential food source for alleviating vitamin A deficiency, particularly in developing countries. With a view to exploit banana peels as a source of valuable components, in the present study, proximate composition, carotenoids, beta‐carotene and some anti‐oxidative enzymes as well as carbohydrate content of selected Indian banana varieties were determined. Karpooravalli cultivar of banana showed the maximum accumulation of carotenoid content in the non‐edible (68 μg g−1 d.w.) portion of banana, while being the second highest in beta‐carotene content (143.12 μg per 100 g). However, Red Banana ranked highest in total carotenoid contents for pulp (4 μg g−1 d.w.) and beta‐carotene was estimated to be the highest in the case of peels (241.91 μg per 100 g) and in pulp (117.2 μg per 100 g). Karpooravalli cultivar of banana is also rich in carbohydrate content in terms of total starch (1786.0 μg g−1 d.w. in peels and 544.85 μg g−1 d.w. in pulp) and sugars (53.53 μg g−1 d.w. in peels and 39.05 μg g−1 d.w. in pulp). The catalase enzyme activity in these peels ranged from 5.66 to 35.57 nmol min−1 mg−1 proteins and was found at a higher level in cultivar Poovan, while the ascorbate peroxidase showed the range of 2.25 to 6.22 μmol min−1 mg−1 proteins. The peels of cultivars Red Banana and Karpooravalli are rich source of bioactive compounds, such as carotenoids (beta‐carotene), anti‐oxidative enzymes and carbohydrate contents.
Effectiveness of fatty acids and their monoglycerides against gram‐negative pathogens
Authors: Altieri Clelia , Bevilacqua Antonio , Cardillo Daniela , Sinigaglia Milena
Published: September 2008

The effectiveness of lauric, myristic and palmitic acids and their monoglycerides against Escherichia coli O157:H7, Yersinia enterocolitica and Salmonella sp. A total of 20 ppm of myristic and palmitic acids and their monolgycerides showed a promising bioactivity (60–80%) against E. coli O157:H7 within 10–24 h; 50 ppm of monolaurin inhibited Y. enterocolitica and E. coli O157:H7 by >90% of control for 96 h; otherwise, 40 ppm of monolaurin and 30–50 ppm of lauric acid reduced Y. enterocolitica growth by >65% of control. The effect of lauric acid and its monoglyceride against Salmonella spp. was moderate (inhibition of approximately 30%). The results of this paper suggested some interesting ideas: the same compound could exert an inhibition or a stimulation of microbial growth; moreover, the studied compounds seemed to act in a reversible manner, as the inhibition of micro‐organism was quite strong within the first 10–24 h and decreased for a prolonged incubation.
British Society of Paediatric Dentistry: a policy document on consent and the use of physical intervention in the dental care of children
Authors: NUNN JUNE , FOSTER MARTIN , MASTER SELINA , GREENING SUE
Published: September 2008

This policy document was prepared by J Nunn, M Foster, S Master and S Greening on behalf of the British Society of Paediatric Dentistry (BSPD). Policy documents produced by the BSPD represent a majority view, based on a consideration of currently available evidence. They are produced to provide guidance with the intention that the policy be regularly reviewed and updated to take account of changing views and developments.
Evaluating evidence of association of bovine tuberculosis in cattle and badgers
Authors: Hone Jim , Donnelly Christl A.
Published: September 2008

Bovine tuberculosis (TB) occurs in cattle and badgers in the UK and control efforts are undertaken to reduce the spread of the disease.

This study evaluates relationships predicted by nine epidemiological two‐host models of disease spread generated by various combinations of density‐dependent, frequency‐dependent and environmental pathogen transmission. The relationships of interest are between measures of TB in cattle and in badgers from 10 sites which were randomly selected to be proactive badger culling sites in the UK Randomized Badger Culling Trial. The data are from the initial badger cull only.

There was most support (Akaike weight = 0·562,R2 = 0·869) for models that predicted a positive linear relationship between density of infectious cattle per square kilometre and the density index of infectious badgers. There was less support (Akaike weight = 0·060) for a model that predicted a positive linear relationship between density of infectious cattle per square kilometre and the proportion of badgers infectious withMycobacterium bovis.A correction to reduce effects of badger carcase storage and an examination of effects of the 2001 foot‐and‐mouth disease epidemic had little impact on estimated relationships.

Synthesis and applications. The results provide support for two‐host disease models of TB in cattle and wildlife such as badgers, although the form of disease transmission cannot be identified clearly by these analyses. The implication of the results is that the best‐fitting models predict that, in the absence of intervention‐related changes in badger behaviour, a reduction in density of infectious badgers should reduce the density of infectious cattle. However, analysis of bait‐marking data collected following experimental badger culls indicated that culling badgers profoundly alters their spatial organization as well as their population density, potentially influencing contact rates. Effective vaccination of badgers, were it to become available, would be expected to reduce the density of infectious badgers without directly affecting their behaviour.
Recruitment of Mycobacterium tuberculosis specific CD4+ T cells to the site of infection for diagnosis of active tuberculosis
Authors: Nemeth J. , Winkler H.‐M. , Zwick R. H. , Rumetshofer R. , Schenk P. , Burghuber O. C. , Graninger W. , Ramharter M. , Winkler S.
Published: September 2008

Context.  Accurate and early diagnosis of active tuberculosis (TB) is problematic as current diagnostic methods show low sensitivity (acid‐fast bacilli smears), are time‐consuming (culture of biological samples) or show variable results [Mycobacterium tuberculosis (MTB)‐specific PCR].

Objectives.  In the course of infection, MTB‐specific T cells clonally expand at the site of infection and may thus be used as diagnostic marker for active disease.

Design.  In this cohort study, the frequency of MTB‐specific, interferon (IFN)‐γ expressing CD4+ T cells obtained from peripheral blood and the site of disease in 25 patients with suspected TB was assessed (n = 11, bronchoalveolar lavage; n = 7, pleural fluid; n = 1, ascites; n = 1, joint fluid; n = 5, cerebrospinal fluid).

Results.  Amongst 15 patients who showed proven active TB infection, a striking increase of MTB‐specific T cells was detected at the site of infection compared with peripheral blood (median increase: 28.5‐fold, range: 7.25–531 fold; median of IFN‐γ‐producing CD4+ T cells from blood: 0.02%, range: 0–0.52%; median of IFN‐γ‐producing CD4+ T cells from the site of infection: 1.81%, range: 0.29–6.55%, P < 0.001).

Main outcome measure.  Recruitment of MTB‐specific T cells to the site of infection yielded a sensitivity of 100% and specificity of 90%, irrespective of the compartment affected.

Conclusions.  The accumulation of MTB‐specific T cells at the site of infection may prove as useful diagnostic marker for an accurate and rapid diagnosis of active TB.
Archaeal RNA polymerase subunits E and F are not required for transcription in vitro, but a Thermococcus kodakarensis mutant lacking subunit F is temperature‐sensitive
Authors: Hirata Akira , Kanai Tamotsu , Santangelo Thomas J. , Tajiri Momoko , Manabe Kenji , Reeve John N. , Imanaka Tadayuki , Murakami Katsuhiko S.
Published: September 2008

All archaeal genomes encode RNA polymerase (RNAP) subunits E and F that share a common ancestry with the eukaryotic RNAP subunits A43 and A14 (Pol I), Rpb7 and Rpb4 (Pol II), and C25 and C17 (Pol III). By gene replacement, we have isolated archaeal mutants of Thermococcus kodakarensis with the subunit F‐encoding gene (rpoF) deleted, but we were unable to isolate mutants lacking the subunit E‐encoding gene (rpoE). Wild‐type T. kodakarensis grows at temperatures ranging from 60°C to 100°C, optimally at 85°C, and the ΔrpoF cells grew at the same rate as wild type at 70°C, but much slower and to lower cell densities at 85°C. The abundance of a chaperonin subunit, CpkB, was much reduced in the ΔrpoF strain growing at 85°C and increased expression of cpkB, rpoF or rpoE integrated at a remote site in the genome, using a nutritionally regulated promoter, improved the growth of ΔrpoF cells. RNAP preparations purified from ΔrpoF cells lacked subunit F and also subunit E and a transcription factor TFE that co‐purifies with RNAP from wild‐type cells, but in vitro, this mutant RNAP exhibited no discernible differences from wild‐type RNAP in promoter‐dependent transcription, abortive transcript synthesis, transcript elongation or termination.
Localization of general and regulatory proteolysis in Bacillus subtilis cells
Authors: Kirstein Janine , Strahl Henrik , Molière Noël , Hamoen Leendert W. , Turgay Kürşad
Published: September 2008

Protein degradation mediated by ATP‐dependent proteases, such as Hsp100/Clp and related AAA+ proteins, plays an important role in cellular protein homeostasis, protein quality control and the regulation of, e.g. heat shock adaptation and other cellular differentiation processes. ClpCP with its adaptor proteins and other related proteases, such as ClpXP or ClpEP of Bacillus subtilis, are involved in general and regulatory proteolysis. To determine if proteolysis occurs at specific locations in B. subtilis cells, we analysed the subcellular distribution of the Clp system together with adaptor and general and regulatory substrate proteins, under different environmental conditions. We can demonstrate that the ATPase and the proteolytic subunit of the Clp proteases, as well as the adaptor or substrate proteins, form visible foci, representing active protease clusters localized to the polar and to the mid‐cell region. These clusters could represent a compartmentalized place for protein degradation positioned at the pole close to where most of the cellular protein biosynthesis and also protein quality control are taking place, thereby spatially separating protein synthesis and degradation.
The RACK1 signal anchor protein from Trypanosoma brucei associates with eukaryotic elongation factor 1A: a role for translational control in cytokinesis
Authors: Regmi Sandesh , Rothberg Karen G. , Hubbard James G. , Ruben Larry
Published: September 2008

RACK1 is a WD‐repeat protein that forms signal complexes at appropriate locations in the cell. RACK1 homologues are core components of ribosomes from yeast, plants and mammals. In contrast, a cryo‐EM analysis of trypanosome ribosomes failed to detect RACK1, thus eliminating an important translational regulatory mechanism. Here we report that TbRACK1 from Trypanosoma brucei associates with eukaryotic translation elongation factor‐1a (eEF1A) as determined by tandem MS of TAP‐TbRACK1 affinity eluates, co‐sedimentation in a sucrose gradient, and co‐precipitation assays. Consistent with these observations, sucrose gradient purified 80S monosomes and translating polysomes each contained TbRACK1. When RNAi was used to deplete cells of TbRACK1, a shift in the polysome profile was observed, while the phosphorylation of a ribosomal protein increased. Under these conditions, cell growth became hypersensitive to the translational inhibitor anisomycin. The kinetoplasts and nuclei were misaligned in the postmitotic cells, resulting in partial cleavage furrow ingression during cytokinesis. Overall, these findings identify eEF1A as a novel TbRACK1 binding partner and establish TbRACK1 as a component of the trypanosome translational apparatus. The synergy between anisomycin and TbRACK1 RNAi suggests that continued translation is required for complete ingression of the cleavage furrow.
Minitags for small molecules: detecting targets of reactive small molecules in living plant tissues using ‘click chemistry’
Authors: Kaschani Farnusch , Verhelst Steven H. L. , Van Swieten Paul F. , Verdoes Martijn , Wong Chung‐Sing , Wang Zheming , Kaiser Markus , Overkleeft Herman S. , Bogyo Matthew , Van Der Hoorn Renier A. L.
Published: September 2008

Small molecules offer unprecedented opportunities for plant research since plants respond to, metabolize, and react with a diverse range of endogenous and exogenous small molecules. Many of these small molecules become covalently attached to proteins. To display these small molecule targets in plants, we introduce a two‐step labelling method for minitagged small molecules. Minitags are small chemical moieties (azide or alkyne) that are inert under biological conditions and have little influence on the membrane permeability and specificity of the small molecule. After labelling, proteomes are extracted under denaturing conditions and minitagged proteins are coupled to reporter tags through a ‘click chemistry’ reaction. We introduce this two‐step labelling procedure in plants by studying the well‐characterized targets of E‐64, a small molecule cysteine protease inhibitor. In contrast to biotinylated E‐64, minitagged E‐64 efficiently labels vacuolar proteases in vivo. We displayed, purified and identified targets of a minitagged inhibitor that targets the proteasome and cysteine proteases in living plant cells. Chemical interference assays with inhibitors showed that MG132, a frequently used proteasome inhibitor, preferentially inhibits cysteine proteases in vivo. The two‐step labelling procedure can be applied on detached leaves, cell cultures, seedlings and other living plant tissues and, when combined with photoreactive groups, can be used to identify targets of herbicides, phytohormones and reactive small molecules selected from chemical genetic screens.
Methyl jasmonate differentially affects tocopherol content and tyrosine amino transferase activity in cultured cells of Amaranthus caudatus and Chenopodium quinoa
Authors: Antognoni F. , Faudale M. , Poli F. , Biondi S.
Published: September 2008

Tocopherols are lipid‐soluble compounds synthesised exclusively by photosynthetic organisms. In this study, in vitro callus cultures were established from two plants that are naturally rich in tocopherols, Amaranthus caudatus and Chenopodium quinoa, in order to examine whether callus cultures were able to produce these compounds at levels comparable to those observed in planta. In both species, cotyledon explants produced the best callus induction and, once established, callus cultures were grown under two different hormonal treatments to check for effects of growth and to induce chloroplast differentiation in the cells. A rapid differentiation of chloroplasts occurred only in C. quinoa cell aggregates grown in the presence of benzyladenine, leading to the production of a homogeneous green callus. In both species, only α‐tocopherol was produced by callus cultures, although levels were much lower than in planta, and the production was not influenced by the hormonal conditions. Interestingly, cell cultures of the two species responded in different ways to methyl jasmonate (MJ). In A. caudatus cultures, treatment with 100 μm MJ increased the production of α‐tocopherol up to fivefold, and the inductive effect was influenced by the hormonal composition of the medium. This increase in α‐tocopherol was associated with a proportional increase in tyrosine aminotransferase (TAT) activity, one of the key enzymes involved in tocopherol biosynthesis. By contrast, in C. quinoa cultures, elicitation with MJ did not have any effect, neither on tocopherol production, nor on TAT activity. These results are discussed in relation to chloroplast differentiation and the interplay between jasmonates and phytohormones.