bluuebunny/crossref_metadata_2025 huggingface dataset:
An improvedin situhybridization method for the detection of fish pathogens
Authors: McCarthy Ú M , Urquhart K L , Bricknell I R
Published: August 2008

A fluorescentin situhybridization (FISH) method was developed for detection of infectious pancreatic necrosis virus (IPNV) in paraffin‐embedded tissues of Atlantic salmon,Salmo salarL. Several methods of probe labelling and detection were evaluated and found unsuitable for FISH because of tissue autofluorescence. Likewise, the use of avidin to detect biotin‐labelled probe was obviated by the presence of endogenous biotin. An existing approach, using digoxigenin (DIG)‐labelled probes and detection by anti‐DIG antibody‐labelled with alkaline phosphatase, was modified to use a fluorescent substrate, 2‐hydroxy‐3‐naphthoic acid‐2′‐phenylanilide phosphate/4‐chloro‐2‐methylbenzene diazonium hemi‐zinc chloride salt (HNPP/Fast Red TR). This improved method allowed sensitive detection of IPNV target, without interference from autofluorescence or endogenous alkaline phosphatase. Furthermore, the reporter produces a discrete, non‐fading signal, which is particularly suitable for analysis by confocal microscopy.
Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction
Authors: Kim D‐G , Bae J‐Y , Hong G‐E , Min M‐K , Kim J‐K , Kong I‐S
Published: August 2008

Vibrio anguillarum, an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum. PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species‐specific fragment (689 bp) from V. anguillarum. Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA, or from six colony‐forming units (CFU) mL−1 of cultured V. anguillarum. However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g−1 tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species‐specific method to detect V. anguillarum in practical situations.
Cell culture detection of microvascular cell death in clinical specimens of human neoplasms and peripheral blood
Authors: Weisenthal L. M. , Patel N. , Rueff‐Weisenthal C.
Published: August 2008

Background.  Angiogenesis studies are limited by the clinical relevance of laboratory model systems. We developed a new method for measuring dead microvascular (MV) cells in clinical tissue, fluid and blood specimens, and applied this system to make several potentially novel observations relating to cancer pharmacology.

Methods.  Dead MV cells tend to have a hyperchromatic, refractile quality, further enhanced during the process of staining with Fast Green and counterstaining with either haematoxylin–eosin or Wright–Giemsa. We used this system to quantify the relative degree of direct antitumour versus anti‐MV effects of cisplatin, erlotinib, imatinib, sorafenib, sunitinib, gefitinib and bevacizumab.

Results.  Bevacizumab had striking anti‐MV effects and minimal antitumour effects; cisplatin had striking antitumour effects and minimal anti‐MV effects. The `nib' drugs had mixed antitumour and anti‐MV effects. Anti‐MV effects of erlotinib and gefitinib were equal to those of sunitinib and sorafenib. There was no detectable VEGF in culture medium without cells; tumour cells secreted copious VEGF, reduced to undetectable levels by bevacizumab, greatly reduced by cytotoxic levels of cisplatin + anguidine, and variably reduced by DMSO and/or ethanol. We observed anti‐MV additivity between bevacizumab and other drugs on an individual patient basis. Peripheral blood specimens had numerous MV cells which were strikingly visualized for quantification with public domain image analysis software using bevacizumab essentially as an imaging reagent.

Conclusions.  This system could be adapted for simple, inexpensive and sensitive/specific detection of tissue and circulating MV cells in a variety of neoplastic and non‐neoplastic conditions, and for drug development and individualized cancer treatment.
Binding of the periodontitis associated bacterium Porphyromonas gingivalis to glycoproteins from human epithelial cells
Authors: Hallén U. , Björkner A. E. , Hallberg E. C.
Published: August 2008

Introduction:  In the present study we examined the ability of the periodontal pathogen Porphyromonas gingivalis to adhere to glycoconjugates on intact cells and to protein preparations of epithelial cells (KB cells).

Methods:  The KB cell protein preparation was separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes by Western blotting. The membranes were used in overlay assays with labeled P. gingivalis. Flow cytometry was used to analyze attachment of bacteria to intact KB cells.

Results:  Glycoconjugate expression on the KB cells and in the protein preparation was confirmed. Binding was detected to several bands on the Western blots. Flow cytometry showed a distinct increase in fluorescence for strain FDC 381. Preincubation of the bacteria with mannose, fucose, N‐acetylglucosamine and N‐acetylgalactosamine inhibited the binding to KB cells by approximately 30% whereas preincubation with N‐acetylneuraminic acid reduced the binding by 60%.

Conclusion:  These results indicate that carbohydrate structures are involved in the binding process of P. gingivalis to oral epithelial cells and that neuraminic acid plays a significant role in the adhesion process.
Rapid identification of oral anaerobic bacteria cultivated from subgingival biofilm by MALDI‐TOF‐MS
Authors: Stîngu C. S. , Rodloff A. C. , Jentsch H. , Schaumann R. , Eschrich K.
Published: August 2008

Introduction:  To facilitate the identification of anaerobes cultivated from periodontal disease, whole cell bacterial identification by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) was evaluated.

Methods:  A total of 84 strains (nine reference strains and 75 recent clinical isolates from 33 patients with aggressive periodontitis) previously identified with phenotypic methods were used. All the references and 10 clinical isolates belonging to the same species as the reference strains were genotypically identified by sequence analysis of the 16S ribosomal RNA gene. All the strains were then analyzed using MALDI‐TOF‐MS.

Results:  The reference strains of anaerobic bacteria used showed characteristic MALDI‐TOF‐MS spectra with peaks between m/z 2000 and up to about m/z 13,000. On visual inspection, the similarity of spectra produced by strains of a single genus could be recognized. Obvious differences between spectra produced by strains of different species were also easily noticed. The reproducibility of the method was proved by the similarity of spectra belonging to the same species. The spectra of the Prevotella intermedia strains identified with MALDI clustered together and clustered separately from the spectra of Prevotella nigrescens, proving that MALDI‐TOF‐MS is an accurate method that is capable of separating these two species. The quality of clustering was characterized by calculating an inconsistency coefficient (Mathworks:/Matlab Reference Manual v2007a/, Statistical toolbox).

Conclusion:  Our results suggest that MALDI‐TOF‐MS might become a useful method for the identification of anaerobic bacteria, especially for those that cannot be readily identified by biochemical analysis. It may become an attractive system even for the routine identification of clinical isolates.
Molecular identification and quantification of bacteria from endodontic infections using real‐time polymerase chain reaction
Authors: Blome B. , Braun A. , Sobarzo V. , Jepsen S.
Published: August 2008

Introduction:  It was the aim of the present study to evaluate root canal samples for the presence and numbers of specific species as well as for total bacterial load in teeth with chronic apical periodontitis using quantitative real‐time polymerase chain reaction (PCR).

Methods:  Forty adult patients with one radiographically documented periapical lesion were included. Twenty teeth presented with primary infections and 20 with secondary infections, requiring retreatment. After removal of necrotic pulp tissue or root canal filling, a first bacterial sample was obtained. Following chemo‐mechanical root canal preparation a second sample was taken and a third sample was obtained after 14 days of intracanal dressing with calcium hydroxide. Analysis by real‐time PCR enabled the quantification of total bacterial counts and of nine selected species.

Results:  Root canals with primary infections harbored significantly more bacteria (by total bacterial count) than teeth with secondary infections (P < 0.05). Mean total bacterial count in the retreatment group was 2.1 × 106 and was significantly reduced following root canal preparation (3.6 × 104) and intracanal dressing (1.4 × 105). Corresponding values for primary infections were: 4.6 × 107, 3.6 × 104, and 6.9 × 104. The numbers of the selected bacteria and their detection frequency were also significantly reduced.

Conclusion:  Root canals with primary infections contained a higher bacterial load. Chemo‐mechanical root canal preparation reduced bacterial counts by at least 95%.
Immunohistochemical localization of Toll‐like receptors 1–10 in periodontitis
Authors: Beklen A. , Hukkanen M. , Richardson R. , Konttinen Y. T.
Published: August 2008

Background/aim:  In periodontitis, bacteria and pathogen‐associated molecular patterns are sensed by Toll‐like receptors (TLRs), which initiate intracellular signaling cascades that may lead to host inflammation. In this study, the expression and distribution of TLRs (TLR‐1 to TLR‐10) were immunohistochemically detected in gingival epithelium and connective tissue.

Methods:  Immunohistochemistry was used for the localization of TLRs in gingival tissue samples from 10 patients with chronic periodontitis and 10 healthy controls; these samples were obtained during routine periodontal flap operations and during extraction operations performed for retained wisdom teeth, respectively. For the evaluation, epithelial layers were stratified to basal, spinous, and superficial layers, and the percentages of TLR‐positive cells were determined.

Results:  Both healthy and periodontitis gingival tissues expressed all TLRs except TLR‐10. In patients with periodontitis, epithelial cells showed increased TLR expression towards the basal layer. Healthy controls showed more variable cellular TLR expression and distribution between the layers of epithelium. In the connective tissue, consistently higher TLR expression was found within the periodontitis group compared to the healthy group.

Conclusions:  For the first time, the cellular expression and distribution of TLR‐1 to TLR‐10 have been studied in periodontitis, indicating that TLR‐1 to TLR‐9 are differentially expressed both in connective tissue and epithelial layers. Except for TLR‐7 and TLR‐8, all the other TLRs showed statistically significant differences between patients with periodontitis and healthy controls, suggesting their involvement in the pathogenesis of periodontitis.
Stimulation of epithelial cell matrix metalloproteinase (MMP‐2, ‐9, ‐13) and interleukin‐8 secretion by fusobacteria
Authors: Gursoy U. K. , Könönen E. , Uitto V.‐J.
Published: August 2008

Background/aims:  Bacterial pathogens involved in periodontal diseases exert their destructive effects primarily by stimulating the host cells to increase their secretion of proinflammatory cytokines and matrix metalloproteinases (MMPs). This study aimed to determine the epithelial cell matrix metalloproteinase and interleukin‐8 (IL‐8) secretion upon exposure to fusobacteria.

Methods:  Eight different oral and non‐oral Fusobacterium strains were incubated with HaCaT epithelial cells. Gelatin zymography and Western blot analysis were performed to detect collagenase 3 (MMP‐13), gelatinase A (MMP‐2), gelatinase B (MMP‐9), and IL‐8 secretion by epithelial cells.

Results:  All Fusobacterium strains, especially Fusobacterium necrophorum ATCC 25286, Fusobacterium nucleatum ATCC 25586, and Fusobacterium varium ATCC 51644, increased MMP‐9 and MMP‐13 secretion. Fusobacterium simiae ATCC 33568, and to a lesser extent F. nucleatum and F. necrophorum, increased epithelial MMP‐2 secretion. F. nucleatum and F. necrophorum also increased IL‐8 secretion. F. varium ATCC 27725, a strain that only weakly stimulated MMP production, strongly increased the IL‐8 production, suggesting that their expression is differently regulated.

Conclusion:  We conclude that the pathogenic potential of fusobacteria may partly result from their ability to stimulate secretion of MMP‐9, MMP‐13, and IL‐8 from epithelial cells.
Single gavage withPorphyromonas gingivalisreduces acute systemic nitric oxide response in mice
Authors: Nemec A. , Pavlica Z. , Šentjurc M. , Crossley D. A. , Jerin A. , Eržen D. , Zdovc I. , Petelin M. , Skalerič U.
Published: August 2008

Introduction: Porphyromonas gingivalis, an important periodontal pathogen, can also induce host responses in distant tissues.P. gingivalisinduces nitric oxide (NO) production in immune system cells and non‐immune system cells, therefore NO might be involved in an acute systemic host response.

Methods: Eighteen female BALB/c mice were perorally inoculated with 108colony‐forming units liveP. gingivalisATCC 33277. Plasma nitrite and nitrate (NOx) and NO production in lungs, aorta, heart, liver, spleen, kidneys, and brain were measured at intervals after inoculation and compared with levels in 11 control animals.

Results: NOx levels were significantly (P = 0.017) lower at 7, 13, and 25 h afterP. gingivalisinoculation. A similar trend in NO production occurred in most tested organs, but never reached statistical significance. The correlation between NOx in plasma and NO in liver was positive (Spearman correlation coefficient = 0.81,P = 0.0025) and marginal for kidney (0.58,P = 0.059).

Conclusion: Single peroral inoculation of mice withP. gingivalisreduces the acute systemic NO response. As NO is important for host defense, the reduction of NO levels after exposure is likely to delay the host response, increasing the chances that infection withP. gingivaliswill become established.
Components of Relationship Quality in Chimpanzees
Authors: Fraser Orlaith N. , Schino Gabriele , Aureli Filippo
Published: August 2008

A novel approach to studying social relationships in captive adult chimpanzees (Pan troglodytes) was taken by using principal components analysis (PCA) to extract three key components of relationship quality from nine behavioural variables. Based on the loadings of the behavioural variables, the components appeared to match previously hypothesized critical aspects of social relationships and were therefore labelled Value, Compatibility and Security. The effects of kinship, sex combination, age difference and time spent together on each of the relationship quality components were analysed. As expected, kin were found to have more valuable, compatible and secure relationships than non‐kin. Female–female dyads were found to be more compatible than male–male or mixed‐sex dyads, whereas the latter were found to be most secure. Partners of a similar age were found to have more secure and more valuable relationships than those with a larger age gap. Individuals that were together in the group for longer were more valuable and more compatible, but their relationships were found to be less secure than individuals that were together in the group for a shorter time. Although some of the results may be unexpected based on chimpanzee socio‐ecology, they fit well overall with the history and social dynamics of the study group. The methods used confer a significant advantage in producing quantitative composite measures of each component of relationship quality, obtained in an objective manner. These findings therefore promote the use of such measures in future studies requiring an assessment of the qualities of dyadic social relationships.
Tail‐Flicking, Tail‐Flagging, and Tail Position in Ungulates with Special Reference to Black‐Tailed Deer
Authors: Stankowich Theodore
Published: August 2008

Testing the functional significance of antipredator behaviors is exceedingly difficult given their potential dual functions (i.e., conspecific warning and pursuit deterrence), and this task is made even more difficult when the same term is used to describe different behavioral patterns by different authors. Tail movements and rump patch exposure in ungulates are well‐documented phenomena, but the functional significance of each has remained elusive. I discuss the confusion in the literature regarding the definition of tail movements, redefine two behavioral patterns and three tail positions, and examine their performance in Columbian black‐tailed deer (Odocoileus hemionus columbianus) in response to an approaching human. ‘Tail‐flicking’, the brief swaying of the tail to one side or the other, was most commonly associated with feeding behavior, and ‘tail‐flagging’, the lateral side‐to‐side movement of the tail in rapid succession, was common during both feeding and alert behaviors. Tail‐flicking and tail‐flagging: (i) were performed by both calm/foraging deer and alarmed deer; (ii) probably lack naturally selected antipredator function but may have the benefit of providing public information to conspecifics about potential danger, and (iii) likely lay at opposite ends of a continuum of tail movements that are the result of agitation and motor reflexes involved in locomotion, where flagging indicates greater agitation than flicking. Deer held their tail ‘erect’ during bouts of rapid flight, including stotting and trotting, and erected their tails sooner after flight during more threatening approaches. Tail erection as a behavioral pattern is an antigravity reflex performed when preparing for locomotion and probably lacks antipredator function. However, tail erection likely facilitated the evolution of bright rump patches in ungulates, which were probably shaped by natural selection to serve multiple antipredator or thermoregulatory functions.
Mating Tactics in Male Grey‐Cheeked Mangabeys (Lophocebus albigena)
Authors: Arlet Małgorzata E. , Molleman Freerk , Chapman Colin A.
Published: August 2008

Reproductive tactics of males can change with individual quality, relatedness and social structure. Here we describe the behaviours of male grey‐cheeked mangabeys towards other males, and females and their offspring (Lophocebus albigena) in relation to male status (high‐ranking/low‐ranking/transient) and group composition in Kibale National Park, Uganda. High‐ranking males had the highest mating success, frequency of loud calls, mate guarding and aggression towards females and males. Only transient males were often observed to be aggressive towards juveniles, while some high‐ranking males provided infant care. Mating tactics of high‐ranking males varied greatly among the five studied groups, probably as a function of the intensity of male–male competition. These results are discussed with regard to the role of male–male competition and behaviours that could affect female mate choice as tactics to obtain reproductive success.
Maternal Epigenetic Transmission of Social Motivation in Birds
Authors: Formanek Laureline , Houdelier Cécilia , Lumineau Sophie , Bertin Aline , Richard‐Yris Marie‐Annick
Published: August 2008

Behavioural development is a complex phenomenon involving interactions between genetic constraints and environmental influences. One of the most potent environmental influences during the ontogeny of the behavioural characteristics of young is played by mothers. In particular, mammalian mothers modulate the social competences of their young, influencing all their future social life. Here, we investigated the influence of the social characteristics of adoptive mother birds (Coturnix coturnix japonica) on the social motivation (origin of every social relationship) of the young they reared. We characterized the social behaviour of standard stock chicks reared by female quail, genetically selected either for low (LSR) or high (HSR) levels of social reinstatement (index of treadmill behaviour which combined the tendency to run towards conspecifics with the tendency to move away from them), to investigate epigenetic transmission of social motivation. Our results show that HSR and LSR adoptive mothers partially transmitted their social characteristics to their young: chicks reared by HSR females presented higher levels of social motivation than chicks reared by LSR females. This maternal influence appeared much clearer in young males than in young females. Our study reveals that, as in mammals, bird mothers influence epigenetically the development of the social behaviour of their young.
Going Out to Mate: Excursion Behaviour of Female Roe Deer
Authors: Lovari Sandro , Bartolommei Paola , Meschi Fiora , Pezzo Francesco
Published: August 2008

Among mammals, female searching behaviour for a mate has been poorly investigated in behavioural studies on sexual selection. The social organization of roe deer and its physiological constraints suggest the existence of searching strategies by females. Using radiotracking and direct observation we studied the spatial behaviour and intersexual interactions of 32 roe does during the rut in two areas of Tuscany, central Italy. Respectively, 55% and 33% of females performed excursions outside their usual home ranges during the peak of the rut (15 Jul.–15 Aug.). Excursions ranged from 0.6 to 3.0 km, if measured in a straight line, and lasted 30–52 h. During excursions, females (1) moved quickly in one direction; (2) met and were courted by a male; (3) copulated with a male and (4) returned to their home ranges after mating. Before starting an excursion, roaming females were observed to approach unsuccessfully males already courting other females. Most likely excursions represent a condition dependent mating tactic adopted by a variable proportion of females and involving active searching behaviour.
Pathogenicity of Chalara fraxinea
Authors: Kowalski T. , Holdenrieder O.
Published: August 2008

The mitosporic fungus Chalara fraxinea (Ascomycota) is associated with dieback of common ash, an emerging disease of Fraxinus excelsior (Oleaceae) in Europe. The pathogenicity of C. fraxinea was demonstrated by field inoculations on young trees.
Analysis of the distribution of Phytophthora cinnamomi in soil at a disease site in Western Australia using nested PCR
Authors: Williams N. , Hardy G. E. St. J. , O’Brien P. A.
Published: August 2008

The oomycete plant pathogen Phytophthora cinnamomi has infected a very large area of native vegetation in the south western corner of Australia. An important aspect of effective disease management depends on being able to accurately map areas of infestation. For this purpose, we have developed a nested polymerase chain reaction (PCR) protocol for the detection of P. cinnamomi in soil. The test uses two sets of primers developed from the rRNA ITS sequences of P. cinnamomi and can detect as little as 1 pg DNA. The degree of sensitivity was reduced with DNA extracted from soil although this depended on the type of soil. Soils with a high organic content, such as eucalypt forest soil and potting mix were more inhibitory than sandy soils. Inhibition by soil DNA could be reduced by the addition of bovine serum albumin and formamide to the reaction. Taq DNA polymerase was very sensitive to inhibitors compared with Tth+ or TaqF1*. In comparison with baiting (0–10% positive samples), nested PCR proved to be a very much more efficient (90–100% positive samples) method for the detection of P. cinnamomi in soil.
European pear rust onJuniperus excelsaL. in south‐western Turkey
Authors: Doğmuş‐Lehtijärvi H. T. , Lehtijärvi A. , Aday A. G.
Published: August 2008

In this study, the frequency of European pear rust (Gymnosporangium fuscum) lesions onJuniperus excelsawas investigated both on seedlings in a nursery and on forest trees in six juniper stands located in south‐western Turkey. In the nursery, lesions occurred on 0.7% of 5510 1‐year‐old seedlings and 29% of 200 2‐year‐old potted seedlings. In each of the juniper forests, 50 trees were chosen within a square of approximately 400 m2in size. In general, rust lesions were common on the trees. On average, 49.0% of the trees had lesions; on 17.3% of the trees, the lesions were located on the trunk, on 43.3% on the branches and on 11.7% on both. In lesion‐bearing trees, the average number of lesions was 1.47–3.59 and the maximum number of lesions per tree were six to 11. Most of the lesions (85.1%) were on branches. There were significant differences between the stands, which can be attributed to different age of the trees, site factors and the presence of alternate hosts. The rust identified from stem lesions in forest trees and from nursery seedlings was European pear rust (G. fuscum), but possible co‐occurrence of otherGymnosporangiumspp. cannot be excluded.
Leisure and ageing: The leisure experiences of older Italians in an Australian community
Authors: Pereira Robert Bernard
Published: August 2008

Background:  A search of the occupational therapy literature revealed a paucity of research on how older adults make use of their leisure time, what meaning these pursuits have to them, and whether their chosen leisure occupations are health enhancing. Occupational therapy literature on how people of culturally and linguistically diverse backgrounds understood leisure was also lacking.

Aim:  This thesis documented a qualitative research study that aimed to discover how Italians aged 65 years and older living in the City of Greater Geelong, Victoria, Australia engaged in leisure occupations, the meanings that were derived from leisure engagement, and whether such subjective experiences of leisure produced health benefits.

Methods and results:  A phenomenological study design was utilised, and data were collected through semistructured interviews and a reflective journal. Participants engaged in numerous meaningful leisure occupations that directly impacted on positive subjective experiences and health outcomes. The subjective experiences found concurred with current research, and were categorised under the three main constructs of leisure theory: ‘intrinsic motivation’, ‘internal control’ and ‘suspension from reality’.

Conclusion:  Leisure was found to promote positive subjective experiences, health benefits and successful ageing for the participants. It is suggested that the subjective experiences categorised under the ‘intrinsic motivation’ construct of leisure theory is the main determinant of positive leisure experiences. Furthermore, culturally meaningful leisure occupations were highlighted, and the ‘Leisure Engagement Model’ is proposed to demonstrate the power of leisure on people's lives and on their sense of health towards continued leisure engagement and health promotion.
Alterations of DNA methylation and clinicopathological diversity of human cancers
Authors: Kanai Yae
Published: August 2008

Alterations of DNA methylation can account for the histological heterogeneity, reflected in the stepwise progression and complex biological characteristics of human cancers, that genetic alterations alone cannot explain. Analysis of DNA methylation status in tissue samples can be an aid to understanding the molecular mechanisms of multistage carcinogenesis. Human cancer cells show a drastic change in DNA methylation status, that is, overall DNA hypomethylation and regional DNA hypermethylation, which results in chromosomal instability and silencing of tumor‐suppressor genes. Overexpression of DNA methyltransferase (DNMT) 1 is not a secondary result of increased cell proliferative activity but may underline the CpG island methylator phenotype of cancers. Splicing alteration of DNMT3B may result in chromosomal instability through DNA hypomethylation of pericentromeric satellite regions. Alterations of DNA methylation are observed even in the precancerous stage frequently associated with chronic inflammation and/or persistent viral infection or with cigarette smoking. Precancerous conditions showing alterations of DNA methylation may generate more malignant cancers. Aberrant DNA methylation is significantly associated with aggressiveness of cancers and poorer outcome of cancer patients. Genome‐wide analysis of DNA methylation status based on array‐based technology may identify DNA methylation profiles that can be used as appropriate indicators for carcinogenetic risk estimation and prognostication.
Activity in the superior colliculus reflects dynamic interactions between voluntary and involuntary influences on orienting behaviour
Authors: Bell Andrew H. , Munoz Douglas P.
Published: August 2008

Performance in a behavioural task can be influenced by both bottom‐up and top‐down processes such as stimulus modality and prior probability. Here, we exploited differences in behavioural strategy to explore the role of the intermediate and deep layers of the superior colliculus (dSC) in covert orienting. Two monkeys were trained on a predictive cued‐saccade task in which the cue predicted the target’s upcoming location with 80% validity. When the delay between cue and target onset was 250 ms, both monkeys showed faster responses to the uncued (Invalid) location. This was associated with a reduced target‐aligned response in the dSC on Valid trials for both monkeys and is consistent with a bottom‐up (i.e. involuntary) bias. When the delay was increased to 650 ms, one monkey continued to show faster responses to the Invalid location whereas the other monkey showed faster responses to the Valid location, consistent with a top‐down (i.e. voluntary) bias. This latter behaviour was correlated with an increase in activity in dSC neurons preceding target onset that was absent in the other monkey. Thus, using the information provided by the cue shifted the emphasis towards top‐down processing, while ignoring this information allowed bottom‐up processing to continue to dominate. Regardless of the selected strategy, however, neurons in the dSC consistently reflected the current bias between the two processes, emphasizing its role in both the bottom‐up and top‐down control of orienting behaviour.
Excitability of pontine startle processing neurones is regulated by the two‐pore‐domain K+ channel TASK‐3 coupled to 5‐HT2C receptors
Authors: Weber Maruschka , Schmitt Angelika , Wischmeyer Erhard , Döring Frank
Published: August 2008

The mammalian startle reflex is a fast response to sudden intense sensory stimuli that can be increased by anxiety or decreased by reward. The cellular integration of sensory and modulatory information takes place in giant neurones of the caudal pontine reticular formation (PnC). The startle reflex is known to be enhanced by 5‐hydroxytryptamine (5‐HT); however, signalling mechanisms that change the excitability of the PnC giant neurones are poorly understood. Possible molecular candidates are two‐pore‐domain K+ (K2P) channels that generate a variable K+ background conductance and control neuronal excitability upon activation of G‐protein‐coupled receptors. We demonstrate by in situ hybridization that the K2P channel TASK‐3 is substantially expressed in PnC giant neurones. Brain slice recordings revealed a corresponding background K+ current in these cells that forms about 30% of the outward current at −30 mV. Inactivation of TASK‐3 at pH 6.4 and by ruthenium red depolarized the cells by about 7 mV and increased the action potential frequency as well as duration. Specific activation of Gαq‐coupled 5‐HT2 receptors with α‐methyl 5‐HT evoked a similar increase of neuronal excitability. Consistently, we measured afferent synaptic inputs from serotonergic raphe neurones and detected 5‐HT2C receptors in PnC giant neurones by immunohistochemistry. Thus, neuronal excitability of PnC giant neurones in vivo is most likely increased by serotonergic projections via the K2P channel TASK‐3.
Pudendal motoneurons of the rat located in separated spinal nuclei possess nicotinic acetylcholine receptors having distinct pharmacological profiles
Authors: Ogier R. , Tribollet E. , Bertrand D. , Raggenbass M.
Published: August 2008

Pudendal motoneurons are located in the ventral horn of the caudal lumbar spinal cord and innervate striated pelvic muscles implicated in sexual and eliminative functions. In rats they are distributed in the dorsomedial (DM) and dorsolateral (DL) nucleus. In male rats, dorsomedial motoneurons innervate the bulbocavernosus, the levator ani and the external anal sphincter, whereas dorsolateral motoneurons control the ischiocavernosus and external urethral sphincter. Using spinal cord slices of young male rats and whole‐cell patch‐clamp recordings, we investigated the sensitivity of pudendal motoneurons to nicotinic cholinergic agonists. Motoneurons were identified following 1,1′‐dilinoleyl‐3,3,3′,3′‐tetramethylindocarbocyanine, 4‐chlorobenzenesulphonate retrograde labelling. In the presence of atropine, both dorsomedial and dorsolateral motoneurons responded to acetylcholine (ACh) by generating a rapidly activating inward current. By using selective nicotinic antagonists and a nicotinic positive allosteric modulator, we found that nicotinic ACh receptors present in dorsomedial and dorsolateral motoneurons display distinct pharmacological profiles. Whereas the former are of the heteromeric type, the latter are predominantly of the α7‐containing type. These data were confirmed by light microscopic autoradiography. In young rats, a ligand for heteromeric nicotinic receptors labelled all laminae of the central grey matter, whereas in the ventral part of the central grey, a ligand selective for α7‐containing nicotinic receptors labelled the DL but not the DM. Dorsolateral and dorsomedial motoneurons innervate two distinct groups of pelvic muscles. A difference in their nicotinic pharmacology may be clinically relevant, as it might allow a selective pharmacological intervention in view of influencing the activity of one or the other set of muscles.
Activation of Fyn tyrosine kinase in the mouse dorsal hippocampus is essential for contextual fear conditioning
Authors: Isosaka Tomoko , Hattori Kotaro , Kida Satoshi , Kohno Tomoko , Nakazawa Takanobu , Yamamoto Tadashi , Yagi Takeshi , Yuasa Shigeki
Published: August 2008

Fyn‐tyrosine‐kinase‐deficient mice exhibit defects in the Morris water maze test and long‐term potentiation (LTP) induction in the hippocampus, and given that LTP has been postulated as the neural basis for memory formation, Fyn may be required for hippocampus‐dependent memory formation. However, how Fyn is involved in the process of memory formation is unclear. To investigate the role of Fyn in hippocampal memory formation, we first tested the behavior of Fyn‐deficient mice by contextual fear conditioning. A mouse was placed in a context and a foot shock was delivered, so that the mouse associated the context with the shock. We found that the freezing response of Fyn‐deficient mice to the context was impaired at 24 h after conditioning. We then measured freezing at 1 h after conditioning, and found that their short‐term contextual fear memory was also impaired. We used Western blotting to examine the mode of Fyn activation in dorsal hippocampal tissue following contextual fear conditioning. Fyn activation peaked as early as 5–10 min after contextual fear conditioning and persisted for at least 40 min. Concomitant increases in tyrosine phosphorylation of several proteins, including NR2B, were also observed, but no increases in tyrosine phosphorylation were observed in Fyn‐deficient mice. Thus, both short‐term and long‐term (24‐h) contextual fear memory were impaired in Fyn‐deficient mice, and Fyn activation in the dorsal hippocampus transiently increased after contextual fear conditioning. These findings strongly suggest that activation of the Fyn signaling pathway is involved in hippocampus‐dependent formation of contextual fear memory.
Morphology and histology of the larynx of the common toad Rhinella arenarum (Hensel, 1867) (Anura, Bufonidae)
Authors: Hermida Gladys N. , Farías Alejandro
Published: August 2008

The structure of the larynx of the toad Rhinella arenarum was exhaustively studied. The laryngeal skeleton consists of three bilaterally symmetrical cartilages: the cricoid and two arytenoids. Internally, each half‐larynx has an anterior and a posterior chamber. The first chamber is delimited by the epithelium covering the arytenoid cartilage and the anterior membrane. The latter consists of fibro‐elastic tissue and contains blood capillaries that, judging by their location and distribution, might serve to maintain vocal cord turgidity. At the level of the cricoid cartilage, two structures are reported here for the first time: the posterodorsal and the anteroventral processes. Both processes are associated with the insertion of the posterior membrane. A cartilaginous rod is located at the free margin of the posterior membrane. This rod appears to support the membrane when the air flows. The distal portion of the larynx communicates with the proximal region of the lung. The epithelium of the laryngeal mucosa contains ciliated cells, goblet cells, secretory cells with short microvilli and neuroendocrine cells immunopositive to PGP 9.5. The results obtained in this study provide new information about the internal organization of the larynx in anurans, which could serve as additional morphological characters for phylogenetic relationships.
Molecular phylogeny of the genus Rotylenchus (Nematoda, Tylenchida) and description of a new species
Authors: Vovlas Nicola , Subbotin Sergei A. , Troccoli Alberto , Liébanas Gracia , Castillo Pablo
Published: August 2008

A description of a new species of plant parasitic nematodes of the genus Rotylenchus from the family Hoplolaimidae is given and a recognition of Rotylenchus jaeni comb. n., previously known as subspecies R. magnus jaeni, as separate species is proposed. Rotylenchus montanus sp. n. is characterized by a hemispherical lip region with six rarely seven annuli, stylet 33–37 µm, female tail rounded, regularly annulated tip with 12–18 annuli and phasmid located 2–9 annuli anterior to anus. Rotylenchus montanus sp. n. is close to species of the monosexual group R. arsenjevi, R. corsicus, R. fragaricus, R. helicus, R. indorobustus and R. neorobustus, by a number of specific characteristics resulting from its specific matrix code: A5, B1, C1, D4, E2, F2, G3, H2, I2, J2, K2. Molecular characterization of R. montanus sp. n. and other Rotylenchus species are provided using D2–D3 expansion segments of 28S and the ITS1 of rRNA genes. The D2–D3 of 28S rRNA and the ITS1–rRNA sequences of R. montanus sp. n. differed in one nucleotide and in 16–20 nucleotides from those of an unidentified Rotylenchus species from Russia, respectively. Molecular analysis of populations of R. magnus and R. jaeni comb. n. demonstrated differences in the D2–D3 and the ITS1–rRNA sequences. These genetic differences together with some minor morphological characters support that both subspecies should be considered as two cryptic sibling species and warranted their elevation to species rank. The result of phylogenetic analysis of Hoplolaimidae for 45 sequences of the D2 and D3 expansion regions of 28S rRNA gene using Bayesian inference analysis under the complex model is presented. Phylogenetic tree of Rotylenchus species represents seven moderate to highly supported lineages. Grouping of Rotylenchus species within other hoplolaimids and analysis of phylogenetic relationships within the genus Rotylenchus using the ITS1 of rRNA gene sequences are also discussed.
Incorporating omega‐3 in the food chain – why, where and how?
Authors: Lunn J. , Buttriss J. L.
Published: August 2008

Summary It has been recognised for decades that high fat intakes are not conducive to good health and, more recently, that the fatty acid profile of the diet (sometimes referred to as fat quality) is also important. Fat intake as a proportion of total food energy has declined over recent years in line with recommendations. Although there have been changes in the types of fats consumed and a fall in the total intake of saturates towards the recommended level, there is still room for improvement. The proportion of energy derived from saturates is still too high in most EU countries. The recommended level is 10% or less of total energy (<11% total food energy) but data show that only two Member States consume 12% or less of total energy as saturates. As the main factor determining serum cholesterol is the amount of saturates in the diet, there are heart health benefits to be gained from achieving a reduction. Meanwhile, research investigating the promise of health benefits linked to increased consumption of n‐6 polyunsaturated fatty acids, monounsaturates and long‐chain n‐3 (omega‐3) polyunsaturates has expanded our knowledge of the health effects of these fatty acids in our diets. Attempts to modify dietary fatty acid profile started with meat and dairy products but developments throughout the food chain have now taken place, such as production of lower fat meats, meat products and spreads; low‐fat milks and dairy products, and a variety of manufactured foods with a reduced fat content and enhanced fatty acid profile. Together these initiatives have led to a fall in saturates intake and improvements in the overall fatty acid profile of our diets, although targets have generally not been met as yet. More recently there have been recommendations to increase our intake of fish, the primary dietary source of long‐chain n‐3 fatty acids. However, the latter has been accompanied by environmental concerns about the safety and sustainability of fish and fish oil supplies. Dietary sources of these long chain n‐3 fatty acids are very limited, oily fish being the only major source. Meanwhile, across Europe, associated with the increased prevalence of obesity and related morbidities, health care costs have soared, exacerbated by the fact that people are living longer although these extra years are often spent in poor health. This has provoked a renewed and reinvigorated search for dietary approaches to health promotion that can potentially be applied to the food supply at the population level. Exploring ways of enhancing the food supply with respect to long‐chain n‐3 fatty acids has been the focus of researchers associated with the EU‐funded Lipgene project. Possible routes include enrichment of meat and meat products through standard animal husbandry techniques, enrichment of milk (either via the cows’ diet or during processing), and more controversially the development of transgenic plants capable of producing long‐chain n‐3 fatty acids, which could be used as a vehicle to introduce these important fatty acids into the food chain.
Regional Governance and Rural Development in Germany: the Implementation of LEADER+
Authors: Böcher Michael
Published: August 2008

This article deals with the currently widely discussed concept of regional governance and investigates the roles this concept actually plays within rural development policy and its implementation in Germany. The main question is whether or not ‘regional governance’ is just a fashionable scientific term. Can we really observe impacts of the regional governance discussion on policies for sustainable regional development in rural areas in Germany? To answer this question the programmatic statements of the EU community initiative LEADER+ are analysed against the background of the most important regional governance criteria. Subsequently, a deeper look into the implementation of LEADER+ in six German regions shows that regional governance still needs some forms of hierarchical co‐ordination by higher levels of state. Some major problems for the implementation of regional governance in LEADER+ as well as some important positive impacts of the programme are identified. These show that, despite all potential problems, the concept of regional governance has the potential to make a significant contribution to the successful development of rural areas.
The Riddle of Ishtar's Shoes: The Religious Significance of the Footprints at ‘Ain Dara from a Comparative Perspective
Authors: THOMAS PAUL BRIAN
Published: August 2008

This essay analyzes the carved footprints in the Iron Age temple at ‘Ain Dara, Syria from a broadly comparative perspective. This methodology is necessitated by the fact that footprint iconography is very rare in ancient Near Eastern art. The diverse examples utilized in this paper to help explicate the footprints at ‘Ain Dara range from the Footprints of the Buddha (Buddhapadas) to the sullied feet of Jesus. Through analysis of footprint iconography in other religious contexts, I have drawn conclusions concerning the form and function of the temple footprints at ‘Ain Dara that have escaped other scholars of the ancient Near East.
Firm foundations? Contingent labour and employers' provision of training in the UK construction sector
Authors: Forde Chris , MacKenzie Robert , Robinson Andrew
Published: August 2008

Based on an original survey of the UK construction industry this article examines the training disadvantages faced by contingent labour. This article addresses two key questions: first, is the provision of training different for directly employed labour and different forms of contingent labour? Second, what structural and organisational factors shape the provision of training to different contingent labour forms? The article finds that fewer firms provide training for agency, self‐employed and subcontract labour, compared to directly employed workers. Training for subcontract labour seems to reflect a longer‐term perspective towards the use of this labour form, while decisions to train agency and self‐employed labour appears to be driven by more short‐term considerations.
Struggles in optimal labelling: how different consumers react to various labels for genetically modified foods
Authors: Teisl Mario F. , Radas Sonja , Roe Brian
Published: August 2008

The labelling of genetically modified (GM) foods is an important policy issue, as consumers' attitudes towards these foods appear to be quite sensitive to information about their potential benefits and risks. Because it is difficult for labels to differ across consumers, differences in reactions to label information could lead to conflicts across consumer groups or to different market outcomes. Using factor and cluster analysis, we uncover three consumer segments with different attitudes to the risks and benefits of GM foods. Our segmentation‐based analysis on consumer reactions to information points out important differences across consumers. Not only did we find differences across segments in how they view the credibility of GM‐related information, we also find differences in how information influences segments' perceptions of the product, and in their likelihood to buy.
The perceived credibility of quality labels: a scale validation with refinement
Authors: Moussa Salim , Touzani Mourad
Published: August 2008

In this paper, we present quality labels as signals that reduce problems that arise under asymmetric information. We propose to closely scrutinize the concept of signal credibility, which is a key determinant of signalling effectiveness. In order to assess the perceived credibility of a quality label, we offer a revisited version of a scale originally proposed by Larceneux. The data used in this paper involve three different labels and were collected using self‐report surveys administered to 602 respondents. Based on findings from a variety of reliability and validity tests, the scale demonstrates good psychometric properties. Both theoretical and managerial implications are discussed, along with limitations and future research directions.