bluuebunny/crossref_metadata_2025 huggingface dataset:
Quantification of immunohistochemistry—issues concerning methods, utility and semiquantitative assessment I
Authors: Walker R A
Published: September 2006

Immunohistochemistry is no longer a technique used only for research but is employed increasingly for diagnosis and for the assessment of therapeutic biomarkers. The latter, in particular, often require a semiquantitative evaluation of the extent of their presence. There are many factors that can affect this that relate to the method: fixation of tissue, duration and type of antigen retrieval, antibody specificity, antibody dilution and detection systems. Other complexities relate to assessment. Different scoring systems are used for either the same or different antigens. Cut‐off levels for assessing whether a tissue is ‘positive’ or ‘negative’ can vary for the same antigen. Whilst there are quality assurance schemes for the methodology that have improved standards of staining, there are no similar schemes that relate to interpretation, although errors here can create as many problems. There have been improvements in automated analysis but availability is limited and it is still predominantly a research tool. In order for quantification of immunohistochemistry to be a reliable and reputable tool, there must be easy to use, reproducible, standardized protocols for assessment which are international. Improvements in automated analysis with wider applicability could lead to standardization.
Cytokeratin profiles of male breast cancers
Authors: Ciocca V , Bombonati A , Gatalica Z , Di Pasquale M , Milos A , Ruiz‐Orrico A , Dreher D , Folch N , Monzon F , Santeusanio G , Perou C M , Bernard P S , Palazzo J P
Published: September 2006

Aims:  The prognostic factors and expression of molecular markers in male breast carcinomas are similar to those in female breast cancers. The identification of distinct cytokeratin (CK) profiles (basal as opposed to luminal cells) helps to identify subsets of tumours with different clinical behaviour. The aim of this study was to investigate CK expression in male breast cancer.

Methods and results:  Thirty‐two cases of male breast cancer were studied. The panel of CKs studied by immunohistochemistry included: 5/6, 14, 17, 18 and 19. Pathological findings and CK expression were analysed in all cases. Histological patterns included ductal carcinoma in situ, invasive ductal carcinoma and mixed patterns. Four cases were positive for CK5/6 and CK14, identifying a basal‐like phenotype. CK17 was negative in all but two cases. All cases expressing either CK5/6 or CK14 were invasive carcinomas of high nuclear and histological grade and were also larger compared with the tumours not expressing CK5/6 and CK14. All tumours except three (also negative for CK5/6) expressed CK18 and CK19. The four basal‐like tumours were negative for Her‐2 expression.

Conclusions:  Male breast carcinomas have a basal‐like phenotype that is similar in frequency to that of female breast carcinomas. The expression of CK5/6 and CK14 identifies a subset of pathologically aggressive male breast cancers.
Effect of emergent aquatic insects on bat foraging in a riparian forest
Authors: FUKUI DAI , MURAKAMI MASASHI , NAKANO SHIGERU , AOI TOSHIKI
Published: September 2006

Riparian zones serve several ecological functions for bats. They provide a source of prey and likely provide favourable structural habitats and shelter from predators. Many studies have shown that bats use the space above streams, ponds or riparian vegetation as feeding habitat. These studies, however, have never distinguished between the effects of habitat structure and prey availability on the foraging activities of bats. Such effects can only be distinguished by an experimental approach. We predicted that bat activity along a stream is influenced by the number of emerged aquatic insects.

We evaluated the response of terrestrial consumers, insectivorous bats, to changes in the abundance of emergent aquatic insects by conducting a manipulative field experiment. In a deciduous riparian forest in Japan, aquatic insect flux from the stream to the riparian zone was controlled with an insect‐proof cover over a 1·2 km stream reach.

We estimated the abundance of emergent aquatic and flying terrestrial arthropods near the treatment and control reaches using Malaise traps. The foraging activity of bats was evaluated in both treatment and control reaches using ultrasonic detectors.

The insect‐proof cover effectively reduced the flux of emergent aquatic insects to the riparian zone adjacent to the treatment reach. Adjacent to the control reach, adult aquatic insect biomass was highest in spring, and then decreased gradually. Terrestrial insect biomass increased gradually during the summer at both treatment and control reaches.

Foraging activity of bats was correlated with insect abundance. In spring, foraging activity of bats at the control reach was significantly greater than at the treatment reach, and increased at both sites with increasing terrestrial insect abundance.

Our result suggests that the flux of aquatic insects emerging from streams is one of the most important factors affecting the distribution of riparian‐foraging bats. As is the case with other riparian consumers, resource subsidies from streams can directly enhance the performance or population density of riparian‐dependent bats. To conserve and manage bat populations, it is important to protect not only forest ecosystems, but also adjacent aquatic systems such as streams.
From plankton to top predators: bottom‐up control of a marine food web across four trophic levels
Authors: FREDERIKSEN MORTEN , EDWARDS MARTIN , RICHARDSON ANTHONY J. , HALLIDAY NICHOLAS C. , WANLESS SARAH
Published: September 2006

Abundant mid‐trophic pelagic fish often play a central role in marine ecosystems, both as links between zooplankton and top predators and as important fishery targets. In the North Sea, the lesser sandeel occupies this position, being the main prey of many bird, mammal and fish predators and the target of a major industrial fishery. However, since 2003, sandeel landings have decreased by > 50%, and many sandeel‐dependent seabirds experienced breeding failures in 2004.

Despite the major economic implications, current understanding of the regulation of key constituents of this ecosystem is poor. Sandeel abundance may be regulated ‘bottom‐up’ by food abundance, often thought to be under climatic control, or ‘top‐down’ by natural or fishery predation. We tested predictions from these two hypotheses by combining unique long‐term data sets (1973–2003) on seabird breeding productivity from the Isle of May, SE Scotland, and plankton and fish larvae from the Continuous Plankton Recorder survey. We also tested whether seabird breeding productivity was more tightly linked to sandeel biomass or quality (size) of individual fish.

The biomass of larval sandeels increased two‐ to threefold over the study period and was positively associated with proxies of the abundance of their plankton prey. Breeding productivity of four seabirds bringing multiple prey items to their offspring was positively related to sandeel larval biomass with a 1‐year lag, indicating dependence on 1‐year‐old fish, but in one species bringing individual fish it was strongly associated with the size of adult sandeels.

These links are consistent with bottom‐up ecosystem regulation and, with evidence from previous studies, indicate how climate‐driven changes in plankton communities can affect top predators and potentially human fisheries through the dynamics of key mid‐trophic fish. However, the failing recruitment to adult sandeel stocks and the exceptionally low seabird breeding productivity in 2004 were not associated with low sandeel larval biomass in 2003, so other mechanisms (e.g. predation, lack of suitable food after metamorphosis) must have been important in this case. Understanding ecosystem regulation is extremely important for predicting the fate of keystone species, such as sandeels, and their predators.
Population limitation of the northern red‐backed vole in the boreal forests of northern Canada
Authors: BOONSTRA RUDY , KREBS† CHARLES J.
Published: September 2006

Across the vast boreal forests of North America, no population cycles inClethrionomysspecies occur. In Eurasia, by contrast, someClethrionomyspopulations of the same species undergo regular 3–5‐year cycles. We examined the effects of nutrients, food, competitors, predators and climate on population limitation in the northern red‐backed vole (Clethrionomys rutilusPallas) in the south‐western Yukon to determine why this difference occurs.

From 1986 to 1996 we added food, reduced large mammal predators and excluded snowshoe hares (Lepus americanusErxleben) from large plots and found that none of these manipulations affected red‐backed vole abundance. Adding nutrients as nitrogen, phosphorus and potassium (NPK) fertilizer had a slight negative effect, probably acting through a reduction in dwarf shrub productivity caused by competition from grasses.

We monitored weasel populations directly through trapping and indirectly through snow tracking. Predation by these vole specialists was irrelevant as a limiting factor most of the time because voles in this area do not reach the densities needed to sustain weasel populations. Other boreal forest mammal and bird predators did not focus on red‐backed voles. However, when red‐backed vole populations increased in the forest andMicrotusvoles also increased in the meadows, weasel populations increased and may have temporarily depressed red‐backed voles in winter.

We monitored one major potential food, white spruce seeds, but seed fall was not related to population changes in red‐backed voles, even after mast years.

We assessed the impact of weather variables, and the average depth of the snow pack during winter (October–March) was correlated directly with vole demography, having both direct effects in that year and delayed effects in the following year.

Our long‐term trapping data (1973–96) indicate thatClethrionomyspopulations fluctuated, with peaks following hare peaks by 2–3 years.

We propose that the key variable limiting these vole populations is overwinter survival, and this is a function of overwinter food from berries produced during the previous summer by dwarf shrubs. These shrubs may be stimulated by abundant moisture from winter snows or by periodic fertilization from large quantities of pellets produced at snowshoe hare peaks.
Distribution of a naturally fluctuating ungulate population among heterogeneous plant communities: ideal and free?
Authors: JONES OWEN R. , PILKINGTON JILL G. , CRAWLEY MICHAEL J.
Published: September 2006

Herbivore distribution is often assumed to follow the ideal free distribution (IFD) model. This assumes that organisms are omniscient about forage quality and availability within the area available to them and are free to move, with negligible cost, throughout this environment. If this were the case we would expect that, at lowest densities, all animals would be found in the best habitat patches, with less desirable habitats being occupied stepwise as population density increases. We test this using data from a naturally fluctuating population of feral Soay sheep.

We show that, although the distribution of individuals is correlated positively with food quality, in line with patterns reported for hill sheep in Scotland, their distribution does not conform to the predictions of the IFD model. We argue that it is the dynamic nature of their food resource that causes this departure from the predictions of the IFD model and make the case that the IFD model, in its unmodified form, is inappropriate for use in modelling distribution among patches containing dynamic resources.
Pili with strong attachments: Gram‐positive bacteria do it differently
Authors: Scott June R. , Zähner Dorothea
Published: September 2006

Bacteria attach to their appropriate environmental niche by using adhesins. To maximize their contact with the environment, adhesins are often present on the ends of long hairlike structures called pili. Recently, attention has focused on pili of Gram‐positive bacteria because they may be vaccine candidates in important human pathogens. These pili differ from the well‐studied pili of Gram‐negative bacteria because their subunits are covalently linked, they do not require specific chaperones for assembly, and the tip protein (likely to be the adhesin) is not required to initiate formation of the pilus structure. In Gram‐positive bacteria, the genes for pili occur in clusters, which may constitute mobile genetic elements. These clusters include the transpeptidase(s) of the sortase family that is/are required for polymerization of the subunit proteins. However, efficient covalent attachment of the completed pilus structure to the cell wall is accomplished, in cases where this has been studied, by the ‘housekeeping’ sortase, which is responsible for attachment to the peptidoglycan of most surface proteins containing cell wall sorting signals. This enzyme is encoded elsewhere on the genome. Because pili of Gram‐positive bacteria have not been extensively investigated yet, we hope that this MicroReview will help to pinpoint the areas most in need of further study.
Identification of the first Oomycete annexin as a (1→3)‐β‐d‐glucan synthase activator
Authors: Bouzenzana Jamel , Pelosi Ludovic , Briolay Anne , Briolay Jérôme , Bulone Vincent
Published: September 2006

(1→3)‐β‐d‐Glucans are major components of the cell walls of Oomycetes and as such they play an essential role in the morphogenesis and growth of these microorganisms. Despite the biological importance of (1→3)‐β‐d‐glucans, their mechanisms of biosynthesis are poorly understood. Previous studies on (1→3)‐β‐d‐glucan synthases from Saprolegnia monoica have shown that three protein bands of an apparent molecular weight of 34, 48 and 50 kDa co‐purify with enzyme activity. However, none of the corresponding proteins have been identified. Here we have identified, purified, sequenced and characterized a protein from the 34 kDa band and clearly shown that it has all the biochemical properties of proteins from the annexin family. In addition, we have unequivocally demonstrated that the purified protein is an activator of (1→3)‐β‐d‐glucan synthase. This represents a new type of function for proteins belonging to the annexin family. Two other proteins from the 48 and 50 kDa bands were identified as ATP synthase subunits, which most likely arise from contaminations by mitochondria during membrane preparation. The results, which are discussed in relation with the possible regulation mechanisms of (1→3)‐β‐d‐glucan synthases, represent a first step towards a better understanding of cell wall polysaccharide biosynthesis in Oomycetes.
Distinct patterns of serum immunoreactivity as evidence for multiple brain‐directed autoantibodies in juvenile neuronal ceroid lipofuscinosis
Authors: Lim M. J. , Beake J. , Bible E. , Curran T. M. , Ramirez‐Montealegre D. , Pearce D. A. , Cooper J. D.
Published: September 2006

Autoantibodies to glutamic acid decarboxylase (GAD65) have been reported in sera from the Cln3–/– mouse model of juvenile neuronal ceroid lipofuscinosis (JNCL), and in individuals with this fatal paediatric neurodegenerative disorder. To investigate the existence of other circulating autoreactive antibodies, we used sera from patients with JNCL and other forms of neuronal ceroid lipofuscinosis (NCL) as primary antisera to stain rat and human central nervous system sections. JNCL sera displayed characteristic patterns of IgG, but not IgA, IgE or IgM immunoreactivity that was distinct from the other forms of NCL. Immunoreactivity of JNCL sera was not confined to GAD65‐positive (GABAergic) neurons, but also stained multiple other cell populations. Preadsorption of JNCL sera with recombinant GAD65 reduced the intensity of the immunoreactivity, but did not significantly change its staining pattern. Moreover, sera from Stiff Person Syndrome and Type I Diabetes, disorders in which GAD65 autoantibodies are present, stained with profiles that were markedly different from JNCL sera. Collectively, these studies provide evidence of the presence of autoreactive antibodies within multiple forms of NCL, and are not exclusively directed towards GAD65.
Molecular chaperons, amyloid and preamyloid lesions in the BRI2 gene‐related dementias: a morphological study
Authors: Lashley T. , Holton J. L. , Verbeek M. M. , Rostagno A. , Bojsen‐Møller M. , David G. , Van Horssen J. , Braendgaard H. , Plant G. , Frangione B. , Ghiso J. , Revesz T.
Published: September 2006

Molecular chaperons or amyloid‐associated proteins (AAPs) are deposited in vascular and parenchymal amyloid lesions in Alzheimer’s disease (AD) and other amyloidoses. AAPs, such as apolipoprotein E (ApoE) or apolipoprotein J (ApoJ) have been strongly implicated in the pathogenesis of AD in vitro and in vivo. Furthermore the possession of the ApoE ∈4 allele is a well‐studied risk factor for AD. In view of the similarities between AD and both familial British dementia (FBD) and familial Danish dementia (FDD), we investigated the presence of AAPs in these two diseases to understand better their role in the general process of amyloidogenesis. Immunohistochemistry for ApoE, ApoJ, serum amyloid P (SAP), alpha‐1‐antichymotrypsin, cystatin C, heparan sulphate proteoglycans, such as agrin, perlecan, syndecans, glypican‐1 and for heparan sulphate glycosaminoglycan (HS GAG) side chains was carried out together with immunohistochemical preparations specific to the amyloid subunits. Significant or extensive staining for ApoE, ApoJ, agrin, glypican‐1 and HS GAG side chains was found in both amyloid (fibrillar) and preamyloid (nonfibrillar) deposits in FBD and FDD. The remaining AAPs, including SAP, were predominantly found in amyloid lesions. Only very weak staining was present in a small proportion of the amyloid lesions using perlecan immunohistochemistry. These findings suggest that the deposition patterns of AAPs in FBD and FDD are mostly similar to those in AD. The presence of AAPs in the preamyloid lesions supports the notion that chaperon molecules may play a role in the early steps of fibrillogenesis.
Lysosome‐associated membrane protein 1 (LAMP‐1) in Alzheimer’s disease
Authors: Barrachina M. , Maes T. , Buesa C. , Ferrer I.
Published: September 2006

Lysosome‐associated membrane protein 1 (LAMP‐1) is a glycoprotein highly expressed in lysosomal membranes. The present study was initiated to test LAMP‐1 mRNA and protein levels in post mortem frontal cortex (area 8) of Alzheimer’s disease (AD) stages I–IIA/B and stages V–VIC of Braak and Braak, compared with age‐matched controls. TaqMan PCR assays and Western blots demonstrated upregulation of LAMP‐1 mRNA and protein in the cerebral cortex in ADVC. In addition, immunohistochemical studies have shown increased LAMP‐1 immunoreactivity in neurones, and in glial cells surrounding senile plaques, in AD cases. Interestingly, LAMP‐1 immunoreactivity has little correlation with phosphorylated tau deposition and neurofibrillary tangles (NFTs), as neurones with NFTs were rarely LAMP‐1 immunoreactive. In contrast, LAMP‐1 expression was enhanced in neurones with granulovacuolar degeneration. Finally, LAMP‐1 occurred in microglia and multinucleated giant cells in one AD case in whom amyloid burden was cleared following βA‐peptide immunization. These findings support the participation of lysosomes in βA‐amyloid and, probably, in hyperphosphorylated tau turnover in AD.
Yellow vein‐affected blackberries and the presence of a novel Crinivirus
Authors: Susaimuthu J. , Tzanetakis I. E. , Gergerich R. C. , Martin R. R.
Published: September 2006

During the last 5 years, blackberry plants in Arkansas and North and South Carolina exhibited virus‐like symptoms of vein yellowing and mosaic, followed in some cases by death. Diagnostic tests for known blackberry viruses failed to identify a causal agent. Double‐stranded RNA was extracted from affected plants and cloned. A new member of the Closteroviridae was identified and designated Blackberry yellow vein associated virus (BYVaV). Molecular and immunological assays have been developed for BYVaV, and examination of plants with symptoms revealed a close association of disease symptoms with the presence of BYVaV, although the virus was also found in symptomless plants. Molecular characterization of isolates from plants exhibiting different degrees of disease severity indicated that sequence diversity is probably not the cause of the observed phenotypic differences.
Response of interspecific Brassica juncea/Brassica rapa hybrids and their advanced progenies to Albugo candida (white blister)
Authors: Gupta K. , Prem D. , Nashaat N. I. , Agnihotri A.
Published: September 2006

Transfer of factors for resistance to white blister disease caused by Albugo candida between Brassica species involving two genotypes each of B. juncea and B. rapa was studied in hybrids. More hybrids were obtained by in vivo than in vitro techniques, although an in vitro phase was a prerequisite for the establishment of in vivo hybrids. Hybrids were identified by PCR‐based inter‐simple sequence repeat (ISSR) markers with both male and female species‐specific bands being identified. There was a positive correlation between disease severity and number of days after sowing (r > 0·93), the highest being towards pod formation and plant maturity at 110 days after sowing. The plants from F2 and BC1 progeny showed higher resistance to A. candida than either of the parents. Plants of B. juncea and B. rapa with high field resistance (disease index < 1·0) were selected from BC2 and F2BC1 generations. The frequency of plants classified as resistant in BC2 progeny ranged from 4·5 to 39·0% in cross‐combinations involving B. juncea genotypes as female parent, compared with 100% in the reciprocal cross involving B. rapa as female parent.
PCR‐based sensitive and specific detection of Pectobacterium atrosepticum using primers based on Rhs family gene sequences
Authors: Park D. S. , Shim J. K. , Kim J. S. , Kim B. Y. , Kang M. J. , Seol Y. J. , Hahn J. H. , Shrestha R. , Lim C. K. , Go S. J. , Kim H. G.
Published: September 2006

A sensitive and specific assay was developed to detect potato blackleg caused by Pectobacterium atrosepticum in potato. Primers PEAF and PEAR from the Rhs family gene homologous to RhsA (cell envelope biogenesis – outer membrane) were used to amplify a 904 bp DNA fragment. PCR was used to detect the pathogen in artificially inoculated potato seed tubers. The PCR product was only produced from 12 isolates of P. atrosepticum from various countries among 36 isolates of other species of Pectobacterium, Pseudomonas, Xanthomonas, as well as Escherichia coli and the soilborne fungus Fusarium oxysporum f.sp. dianthi.
Analysis of population structure of Erysiphe necator using AFLP markers
Authors: Núñez Y. , Gallego J. , Ponz F. , Raposo R.
Published: September 2006

An analysis of the population structure of Erysiphe necator from Spain was carried out to determine if population structure of grapevine powdery mildew is partitioned by origin of primary inoculum, i.e. overwintering ascomata on grapevine bark and mycelia in dormant buds. Ten of the 31 isolates collected were from flag shoots, i.e. shoots covered with sporulating mycelia that arise from dormant infected buds. Genetic variation was assessed using 61 AFLP markers. Cluster analysis revealed two distinct groups with 63% genetic similarity. Principal coordinates analysis (pccorda) produced a similar distribution of isolates in two groups. Isolates were not well identified within their groups by the origin of primary inoculum. Analysis of molecular variance (amova) determined that the origin of primary inoculum was not a significant component in the genetic variation in the population of E. necator. It was concluded that these two subpopulations are not separated by the type of primary source of inoculum.
Resistance in water yam (Dioscorea alata) cultivars in the French West Indies to anthracnose disease based on tissue culture‐derived whole‐plant assay
Authors: Onyeka T. J. , Pétro D. , Ano G. , Etienne S. , Rubens S.
Published: September 2006

Reactions of 60 water yam (Dioscorea alata) cultivars to three isolates of the yam anthracnose fungal pathogen (Colletotrichum gloeosporioides) were evaluated using tissue culture‐derived whole‐plant assay. Three disease parameters: single score on a scale of 0–6 at the seventh day after inoculation (SD7); area under the disease progress curve (AUDPC); and disease progress rate (Rd) were compared, and cultivars were classified into disease‐response groups using a rank‐sum method based on AUDPC scores for the two most virulent isolates. A wide range of variation in resistance of the D. alata cultivars, and significant effects of pathogen isolate and isolate–cultivar interactions, were observed for all disease parameters. The three disease parameters were positively correlated; however, four cultivars showed great dispersions from the regression lines for comparisons of SD7 with the multiple assessments based AUDPC and Rd. The 60 cultivars were separated into resistant (n = 12), moderately resistant (n = 19), moderately susceptible (n = 18) and susceptible (n = 11) groups. The potential of the tissue culture‐derived whole‐plant assay to resistance breeding programmes and further understanding of the yam anthracnose pathosystem is discussed.
Genetic, phenotypic and pathogenic diversity among xanthomonads isolated from pistachio (Pistacia vera) in Australia
Authors: Marefat A. , Scott E. S. , Ophel‐Keller K. , Sedgley M.
Published: September 2006

Repetitive extragenic palindromic polymerase chain reaction (rep‐PCR), sequencing of the 16S−23S rDNA internal transcribed spacer (ITS), biochemical and physiological tests, the Biolog microplate system, polyacrylamide gel electrophoresis (PAGE) of whole‐cell proteins, and pathogenicity tests were used to characterize variability among xanthomonads isolated from pistachio trees suffering from bacterial dieback in four regions of Australia. ITS sequencing and rep‐PCR revealed two distinct genotypes among the strains. The ITS sequencing suggested that the pistachio strains were closely related to Xanthomonas translucens pathovars, in particular X. translucens pv. poae. Results of physiological and biochemical tests, as well as Biolog microplate analysis and protein profiling, confirmed the existence of two groups. Furthermore, pathogenicity and host‐range studies indicated that the two groups were biologically different. There was an association between the two groups and the geographical origin of the strains.
Interaction between Pseudomonas savastanoi pv. savastanoi and Pantoea agglomerans in olive knots
Authors: Marchi G. , Sisto A. , Cimmino A. , Andolfi A. , Cipriani M. G. , Evidente A. , Surico G.
Published: September 2006

Results of a survey of olive knot disease in central Italy in 2002 and 2003 showed that Pantoea agglomerans was found associated with the pathogen Pseudomonas savastanoi pv. savastanoi (Ps. savastanoi) in 70% of the olive knots examined. Pathogenicity tests in which these two bacteria were co‐inoculated on the stems of 1‐year‐old olive plants at ratios of 1:1, 1:100 and 100:1 showed that the growth of P. agglomerans was apparently aided by the presence of an actively growing population of Ps. savastanoi. At the same time, however, a dominant population of P. agglomerans at the inoculation site tended to depress the growth of Ps. savastanoi, probably because of competition for space and nutrients between these bacteria and by means of antibiotic production by P. agglomerans. In some cases the association of P. agglomerans, which in culture was found to produce indole‐3‐acetic acid but not cytokinins, with Ps. savastanoi resulted in an increase in the size of knots. This boosting effect of P. agglomerans on proliferation was probably due to the release of IAA by this bacterium at the inoculation sites.
Review of the South American genus Eurotettix Bruner (Orthoptera, Acridoidea, Melanoplinae)
Authors: CIGLIANO MARIA MARTA
Published: September 2006

Abstract The South American grasshopper genus Eurotettix (Melanoplinae, Dichroplini) from north‐eastern Argentina, southern Brazil and eastern Paraguay is revised. Cladistic analysis of the morphological and karyotypic characters indicates that the monophyly of Eurotettix is supported only weakly. Thus, Eurotettix is divided into two genera, and Dichromatos gen.n. is described for Eurotettix corupa Carbonell & Mesa, E. lilloanus Liebermann (type species), E. montanus Carbonell & Mesa and E. schrottkyi Rehn. Twelve species are recognized for Eurotettix with six described as new: E. concavus sp.n., E. latus sp.n., E. procerus sp.n., E. brevicerci sp.n., E. similraphael sp.n. and E. bugresensis sp.n. Keys to the species of the genera and a review of defining morphological characters are provided.
A mitochondrial genome phylogeny of Diptera: whole genome sequence data accurately resolve relationships over broad timescales with high precision
Authors: CAMERON STEPHEN L. , LAMBKIN CHRISTINE L. , BARKER STEPHEN C. , WHITING MICHAEL F.
Published: September 2006

Abstract Mitochondrial genomes provide a promising new tool for understanding deep‐level insect phylogenetics, but have yet to be evaluated for their ability to resolve intraordinal relationships. We tested the utility of mitochondrial genome data for the resolution of relationships within Diptera, the insect order for which the most data are available. We sequenced an additional three genomes, from a syrphid, nemestrinid and tabanid, representing three additional dipteran clades, ‘aschiza’, non‐heteroneuran muscomorpha and ‘basal brachyceran’, respectively. We assessed the influence of optimality criteria, gene inclusion/exclusion, data recoding and partitioning strategies on topology and nodal support within Diptera. Our consensus phylogeny of Diptera was largely consistent with previous phylogenetic hypotheses of the order, except that we did not recover a monophyletic Muscomorpha (Nesmestrinidae grouped with Tabanidae) or Acalyptratae (Drosophilidae grouped with Calliphoridae). The results were very robust to optimality criteria, as parsimony, likelihood and Bayesian approaches yielded very similar topologies, although nodal support varied. The addition of ribosomal and transfer RNA genes to the protein coding genes traditionally used in mitochondrial genome phylogenies improved the resolution and support, contrary to previous suggestions that these genes would evolve too quickly or prove too difficult to align to provide phylogenetic signal at deep nodes. Strategies to recode data, aimed at reducing homoplasy, resulted in a decrease in tree resolution and branch support. Bayesian analyses were highly sensitive to partitioning strategy: biologically realistic partitions into codon groups produced the best results. The implications of this study for dipteran systematics and the effective approaches to using mitochondrial genome data are discussed. Mitochondrial genomes resolve intraordinal relationships within Diptera accurately over very wide time ranges (1–200 million years ago) and genetic distances, suggesting that this may be an excellent data source for deep‐level studies within other, less studied, insect orders.
Differential expression of pectate lyase during ethylene‐induced postharvest softening of mango (Mangifera indica var. Dashehari)
Authors: Chourasia Amita , Sane Vidhu A. , Nath Pravendra
Published: September 2006

Fruit ripening is characterized by progressive depolymerisation of pectic and hemicellulosic polysaccharides of the cell wall. Several cell‐wall hydrolases have been studied and their role implicated in the softening of various fruits. However, studies related to pectate lyases (PL), which degrade pectins through β‐elimination of glycosidic linkages, have been very limited in fruit tissue. We have cloned a PL homologue, MiPel1 from ripening mango (Mangifera indica var. Dashehari), which shows sequence similarity to higher plant PL genes. The 1.7‐kb complementary DNA of MiPel1 (accession number AY987389) encodes a putative protein of 437 amino acids, which possesses all the motifs and active site residues conserved in other PL. A progressive increase in transcript accumulation was observed during ripening but expression was delayed significantly in 1‐methylcyclopropene‐treated and control fruits kept in air without exogenous ethylene. The expression was specific to fruits and triggered only during ripening. No transcript was detected in other tissues such as leaf, stem and flower and during development of the fruit. The increase in transcript accumulation of MiPel1 during ripening was associated with an increase in the Ca+2‐dependent PL activity and pectin solubilization. It is proposed that expression of PL may be closely associated with pectin degradation during ripening and play an important role in mango softening.
Cadmium partitioning and gene expression studies in Nicotiana tabacum and Nicotiana rustica
Authors: Bovet Lucien , Rossi Luca , Lugon‐Moulin Nicolas
Published: September 2006

To better understand the differences in cadmium (Cd) uptake, partitioning and gene regulation between Nicotiana tabacum and Nicotiana rustica, we compared these two species for root and leaf Cd concentrations after different Cd exposures, 109Cd root‐to‐shoot transport, Cd tolerance as well as differential gene expression in roots exposed or not to CdCl2 using reverse transcriptase–PCR (RT‐PCR). When grown in 1 μM CdCl2 for 7 days, N. rustica exhibited higher root and lower leaf Cd contents than N. tabacum. Data were confirmed by radiolabeling experiments, which further showed that some 109Cd accumulated in the distal part of lateral roots in N. rustica. Visual inspection of leaves suggested that N. rustica was somewhat more tolerant to high Cd exposure (50 μM CdCl2) compared with N. tabacum. At such a high Cd concentration, Cd toxic effects on N. tabacum leaves were apparently not directly related to the homeostasis of Fe and Mn. However, the Zn levels were different in N. rustica compared with N. tabacum in absence and presence of Cd treatments. Root growth experiments revealed that N. tabacum, but not N. rustica, root length was reduced in bactoagar medium containing 20 μM CdCl2. Complementary DNA microarrays were used as a screening approach to demonstrate by RT‐PCR that some gene products were differentially regulated by Cd in N. rustica and in N. tabacum. In addition, “NtIRT1,”NtMTP1a, “NtHMA3” and “NtNAS3” were inducible by Cd in N. tabacum. Interestingly “NtIRT1” and NtMTP1a were differently expressed between the two species. Our results suggest different pathways for Cd sequestration and transport between these two species.
Expression profiling of genes related to starch synthesis in rice leaf sheaths during the heading period
Authors: Hirose Tatsuro , Ohdan Takashi , Nakamura Yasunori , Terao Tomio
Published: September 2006

The stems (leaf sheaths and culms) of rice plants (Oryza sativa L.) accumulate high levels of starch before heading, which is subsequently remobilized after heading to provide a carbon source for grain filling. To elucidate the molecular mechanism of starch synthesis in rice leaf sheath, a comprehensive expression analysis was conducted on the gene families encoding starch synthesis–related enzymes, ADP‐glucose pyrophosphorylase (EC 2.7.7.27), starch synthase (EC 2.4.1.21) and branching enzyme (EC 2.4.1.18). The changes in the activities of these enzymes in leaf sheaths during the heading period were shown to be accompanied by a coordinated change in the transcript levels of particular members of the corresponding gene families. A similar change before and after heading was also found in the messenger RNA level of GPT2, one of the two genes encoding for a plastidial glucose‐6‐phosphate/phosphate translocator, suggesting that the capacity of both starch synthesis and its substrate import is under coordinated transcriptional regulation. The members of the gene families predominantly expressed in leaf sheaths are mostly different from those actively expressed in the developing endosperm. The time course for the transcript level of some of these genes appears to correlate to hexose level in the leaf sheaths, suggesting that hexose levels may be a controlling factor in their expression.
Enhanced expression of genes for ACC synthase, ACC oxidase, and NAC protein during high‐temperature‐induced necrosis of young inflorescences of Cymbidium
Authors: Mita Satoru , Henmi Ryuya , Ohno Hajime
Published: September 2006

Growing Cymbidium under high‐temperature conditions (25–30°C) results in the necrosis of young inflorescences. An increase in the evolution of ethylene was correlated with the necrosis. To study the molecular aspects of high‐temperature‐induced necrosis of Cymbidium floral buds, we isolated complementary DNA (cDNA) clones for proteins that are likely to be involved in the biosynthesis of ethylene during high‐temperature‐induced necrosis of young inflorescences, namely, 1‐aminocyclopropane‐1‐carboxylic acid (ACC) synthase (CyACS1) and ACC oxidase (CyACO1). In addition, a cDNA (CyNAC1) encoding an NAC protein whose expression is modulated during high‐temperature treatment was isolated by differential display. High levels of expression of CyACS1, CyACO1 and CyNAC1 were observed in the necrotic inflorescences of wild‐type Cymbidium at high temperatures. Bud necrosis was not observed in the mericlone mutant (nhn, non–high‐temperature‐induced necrosis) of Cymbidium. Ethylene evolution was lower in nhn than in wild‐type, but application of exogenous ACC or ethephon to the young inflorescences of nhn restored the high‐temperature necrosis response. Expression of CyACS1, CyACO1 and CyNAC1 did not increase with high‐temperature treatment in the nhn mutant. Expression levels of CyACS1, CyACO1 and CyNAC1 in necrotic inflorescences of nhn treated with 5.0 mM ACC were much lower than in necrotic inflorescences of wild‐type at high temperatures, but CyACS1 and CyNAC1 were stimulated by ACC treatment. These results suggest that ethylene is involved in high‐temperature necrosis of young inflorescences of Cymbidium and that an NAC protein may be involved in the regulatory mechanisms of genes that are regulated during necrosis.
Effects of isoflurane, pentobarbital, and urethane on apoptosis and apoptotic signal transduction in rat kidney
Authors: Aravindan N. , Cata J. P. , Hoffman L. , Dougherty P. M. , Riedel B. J. , Price K. J. , Shaw A. D.
Published: September 2006

Background:  Renal cell apoptosis contributes significantly to the pathogenesis of acute renal failure. Anesthetic agents have been shown to modulate apoptotic signal transduction in various tissues. We examined the effects of 6 h of different general anesthetic techniques on renal cell apoptosis in rat kidneys.

Methods:  Twenty‐one male Sprague–Dawley rats were randomly allocated into four groups: (i) control, non‐anesthetized rats (n= 3) and rats anesthetized with (ii) inhaled isoflurane (n= 6), (iii) intraperitoneal pentobarbital (n= 6), and (iv) intraperitoneal urethane (n= 6). Animals were sacrificed 6 h after the induction of anesthesia.

Results:  Apoptosis was assessed by terminal deoxynucleotidyl transferase‐fluorescein end‐labeling analysis. RNA was extracted from the left kidney to probe cDNA microarrays. Gene expression was measured as a percentage of glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) and subsequently confirmed using reverse transcriptase‐polymerase chain reaction (RT‐PCR). Compared with the control (no anesthesia), urethane significantly (P < 0.001) induced apoptosis in both the renal cortex and medulla. Isoflurane significantly (P < 0.001) inhibited apoptosis in the medulla. Microarray analysis revealed that urethane up‐regulated more (74) genes than pentobarbital (16) and isoflurane (10). Isoflurane down‐regulated more genes (85) than pentobarbital (74) and urethane (12). These anesthetic‐induced modulations were significant (P < 0.05) for 60 isoflurane‐, 30 pentobarbital‐ and 4 urethane‐modulated genes.

Conclusion:  Our results suggest that general anesthetic drugs have an effect on renal cell apoptosis and apoptotic signal transduction, and thus may potentially affect the risk of subsequent acute renal failure.
Post‐void residual volume in labor: a prospective study comparing parturients with and without epidural analgesia
Authors: Weiniger C. F. , Wand S. , Nadjari M. , Elchalal U. , Mankuta D. , Ginosar Y. , Matot I.
Published: September 2006

Background:  This prospective, non‐randomized study compared post‐void residual volume in laboring and postpartum women with or without epidural analgesia.

Methods:  The study was conducted over 1 year with institutional review board approval. Parturients were recruited in early labor and self‐selected to either the study (with epidural) or control (without epidural) group. Post‐void residual volume was compared between groups, using transabdominal ultrasound during labor, and on postpartum day 1 and 2. Main outcome measure was intrapartum residual bladder volume.

Results:  Thirty patients were recruited to each group. During labor, residual bladder volume was significantly larger in the epidural group compared with the non‐epidural group [median (range)] 240 (12–640), ml vs. 45 (13–250) ml, respectively, P < 0.001], but was similar on postpartum day 1 and 2. Twenty‐five (83%) women with epidural analgesia required bladder catheterization during labor vs. one (3.3%) without (P < 0.0001).

Conclusion:  The greater post‐void residual volume and increased inability to void in parturients with epidurals suggests that epidural analgesia plays a role in intrapartum urinary retention.
The effects of pre‐operative dexmedetomidine infusion on hemodynamics in patients with pulmonary hypertension undergoing mitral valve replacement surgery
Authors: But A. K. , Ozgul U. , Erdil F. , Gulhas N. , Toprak H. I. , Durmus M. , Ersoy M. O.
Published: September 2006

Background:  The aim of this study was to investigate the effects of pre‐operative dexmedetomidine infusion on hemodynamics in patients with pulmonary hypertension undergoing mitral valve replacement surgery.

Methods:  Patients were randomly divided into placebo (group P, n= 16) and dexmedetomidine (group D, n= 16) groups. In group D, a 1 μg/kg bolus dose of dexmedetomidine was administered 10 min before the induction of anesthesia, followed by a 0.4 μg/kg/h infusion until the surgical incision. Anesthesia was induced with lidocaine (1 mg/kg), midazolam (0.2 mg/kg) and fentanyl (5 μg/kg) in both groups. Anesthesia was maintained with 0.5% isoflurane and fentanyl depending on the hemodynamic situation. The hemodynamic values during the investigation were obtained.

Results:  In group D, the mean arterial pressure (MAP), mean pulmonary arterial pressure (MPAP) and pulmonary capillary wedge pressure (PCWP) were decreased effectively in comparison with the values in the placebo group (P < 0.05), and there was an attenuation in the increase in the systemic vascular resistance index (SVRI) and pulmonary vascular resistance index (PVRI) at the post‐sternotomy period.

Conclusions:  The pre‐operative administration of the α2‐agonist dexmedetomidine decreases the fentanyl requirement and attenuates the increase in SVRI and PVRI at the post‐sternotomy period relative to the baseline levels, and decreases effectively MAP, MPAP and PCWP in comparison with the values in the placebo group, in patients with pulmonary hypertension undergoing mitral valve replacement surgery.
Inhaled iloprost in eight heart transplant recipients presenting with post‐bypass acute right ventricular dysfunction
Authors: Theodoraki K. , Tsiapras D. , Tsourelis L. , Zarkalis D. , Sfirakis P. , Kapetanakis E. , Alivizatos P. , Antoniou T.
Published: September 2006

Background:  During heart transplantation, weaning from cardiopulmonary bypass may be particularly laborious as a result of superimposed acute right ventricular dysfunction in the setting of pre‐existing pulmonary hypertension. Research in recent years has focused on inhaled vasodilatory treatment modalities which selectively target the pulmonary circulation.

Methods:  We present a series of eight patients in whom inhaled iloprost, a synthetic prostacyclin analog, was used to treat pulmonary hypertension and right ventricular dysfunction detected by transesophageal echocardiography during a heart transplant procedure. In addition to conventional inotropic support, 20 μg of inhaled iloprost was administered via nebulized aerosol for a 20‐min period. Complete sets of hemodynamic measurements were obtained before inhalation and during and after cessation of the inhalation period.

Results:  Inhaled iloprost decreased the transpulmonary gradient at the end of the inhalation period relative to baseline (8.2 ± 1.6 mmHg vs. 11.2 ± 0.9 mmHg, P < 0.05). The mean pulmonary artery pressure to systemic artery pressure ratio decreased over this period (0.24 ± 0.07 vs. 0.44 ± 0.09, P < 0.05). A statistically significant decrease in the pulmonary vascular resistance to systemic vascular resistance ratio was also observed (0.10 ± 0.02 vs. 0.19 ± 0.02, P < 0.05). Improved indices of right ventricular function were observed in echocardiographic monitoring.

Conclusion:  During heart transplantation procedures, episodes of pulmonary hypertension can be successfully treated with inhaled iloprost administration, without untoward side‐effects or significant systemic impact.
Pine mistletoe (Viscum albumssp.austriacum) contributes to Scots pine (Pinus sylvestris) mortality in the Rhone valley of Switzerland
Authors: Dobbertin M. , Rigling A.
Published: September 2006

In recent years unusual high mortality of Scots pine (Pinus sylvestris) has been observed in the Swiss Rhone Valley. The exact causes, however, are not known. At a 2‐ha monitoring plot, tree mortality and crown condition have been monitored since 1996. Between 1996 and 2004, 59% of the Scots pines died, most of them following the drought periods 1996–1998 and 2003–2004, while only 15% of the deciduous trees died. Crown transparency, needle discolouration, dead branch percentage, mistletoe (Viscum albumssp.austriacum) rating,Tomicussp. shoot feeding, male flowering effect, tree stem diameter, crown shading and social tree class assessed in 1998 were used in a logistic regression model to predict tree mortality. Crown transparency, mistletoe rating and percentage of dead branches were found significant in the model and the probability of tree mortality increased with increasing rankings of these parameters. Needle discolouration could be used to substitute ‘dead branch percentage’ as predictor. While crown transparency increased with mistletoe rating, for trees in the same transparency class, trees with medium and heavy mistletoe infection were two to four times more likely to die than trees with no or only low mistletoe infection. For the surviving trees we found that trees with mistletoes showed a significantly higher increase in transparency in the year following a drought than trees without, while in a drought year the opposite was true. At the beginning of the observations no significant differences in transparency had been found between the trees with and without mistletoe. However, by the end of the observation period trees with mistletoe had significantly higher crown transparency. We conclude that mistletoe infection can be considered as both a predisposing factor for tree death, by increasing needle loss following drought and a contributing factor by increasing water stress during drought.