bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of the complete porcine MSTN gene and expression levels in pig breeds differing in muscularity
Authors: Stinckens A. , Luyten T. , Bijttebier J. , Van den Maagdenberg K. , Dieltiens D. , Janssens S. , De Smet S. , Georges M. , Buys N.
Published: September 2008

Myostatin (MSTN), a transforming growth factor β superfamily member, is an essential factor for the growth and development of muscle mass. The protein functions as a negative regulator of muscle growth and is related to the so‐called double‐muscling phenotype in cattle, where a series of mutations renders the gene inactive. One particular breed of pigs, the Belgian Piétrain, also shows a heavily muscled phenotype. The similarity of muscular phenotypes between the double‐muscled cattle and Piétrain pigs indicated that MSTN may be a candidate gene for muscular hypertrophy in pigs. In this study, we sequenced and analysed the complete MSTN gene from 45 pigs of five different breeds, including the heavily muscled Piétrain breed at one extreme and the Meishan and Wild boar breeds at the other extreme. In total, 7626 bp of the porcine MSTN gene were sequenced, including the 5′ and 3′ UTR. Fifteen polymorphic loci were found, three of which were located in the promoter region, five in intron 1 and seven in intron 2. Most mutations were found when comparing the obtained MSTN sequence with porcine MSTN sequences already published. However, one polymorphism located at position 447 of the porcine MSTN promoter had a very high allele frequency in the Piétrain pig breed and disrupted a putative myocyte enhancer factor 3 binding site. Real‐time PCR using Sybr Green showed that this mutation was associated with expression levels of the MSTN gene in m. longissimus dorsi at an age of 4 weeks.
Ancestral matrilineages and mitochondrial DNA diversity of the Lidia cattle breed
Authors: Cortés O. , Tupac‐Yupanqui I. , Dunner S. , García‐Atance M. A. , García D. , Fernández J. , Cañón J.
Published: September 2008

To clarify the genetic ancestry and the mitochondrial DNA (mtDNA) diversity of the Lidia cattle breed, a 521‐bp D‐loop fragment was sequenced in 527 animals belonging to 70 herds distributed across 29 lineages. The mtDNA diversity recorded was similar to that seen for Middle Eastern breeds and greater than that recorded for the majority of European breeds. Haplotype T3 was the most common (81%), followed by the African T1 haplotype (17%); very low frequencies were recorded for haplotypes T and T2. The results agree with there being two major ancestral lines for the Lidia breed, European and African, similar to that seen for other Mediterranean breeds. A wide range of variation in haplotype frequencies was seen between the examined lineages. Haplotype T3 was present in all those analysed; in five it was the only one present, and in only one lineage (Miura) was its frequency lower than that of T1. T1*, a haplotype reported in Criollo breeds and to date in only a single European breed (the Retinta breed from Spain), was found in a single animal belonging to the Concha y Sierra lineage. Network analysis of the Lidia breed revealed the presence of two major haplotypes: T3 and T1. The Lidia breed appears to be more closely related to prehistoric Iberian and Italian than to British aurochs.
Albinism in the American mink (Neovison vison) is associated with a tyrosinase nonsense mutation
Authors: Anistoroaei R. , Fredholm M. , Christensen K. , Leeb T.
Published: September 2008

Albino phenotypes are documented in various species including the American mink. In other species the albino phenotypes are associated with tyrosinase (TYR) gene mutations; therefore TYR was considered the candidate gene for albinism in mink. Four microsatellite markers were chosen in the predicted region of the TYR gene. Genotypes at the markers Mvi6025 and Mvi6034 were found to be associated with the albino phenotype within an extended half‐sib family. A BAC clone containing Mvi6034 was mapped to chromosome 7q1.1‐q1.3 by fluorescent in situ hybridization. Subsequent analysis of genomic TYR sequences from wild‐type and albino mink samples identified a nonsense mutation in exon 1, which converts a TGT codon encoding cysteine to a TGA stop codon (c.138T>A, p.C46X; EU627590). The mutation truncates more than 90% of the normal gene product including the putative catalytic domains. The results indicate that the nonsense mutation is responsible for the albino phenotype in the American mink.
Associations between the 11‐bp deletion in the myostatin gene and carcass quality in Angus‐sired cattle
Authors: Gill J. L. , Bishop S. C. , McCorquodale C. , Williams J. L. , Wiener P.
Published: September 2008

An 11‐bp deletion in the bovine myostatin (MSTN) gene was identified as the causative mutation for the double‐muscling phenotype in Belgian Blue and Asturiana cattle. More recently, this mutation was also identified in the South Devon breed of cattle, in which it has been found to be associated with a general increase in muscle mass. The present study found that the mutant allele was also segregating in a commercial population of Scottish Aberdeen Angus beef cattle. The mutation was found at a low frequency (0.04) with no animals homozygous for the mutation in the sample population (536 animals). The effects of this mutation on various carcass traits of economic interest were then tested. We found that the mutation significantly increased carcass weight, sirloin weight, hindquarter weight, muscle conformation score and eye muscle area, but had no effect on the fat traits.
Family treatment of childhood anxiety
Authors: Maid Rachel , Smokowski Paul , Bacallao Martica
Published: September 2008

Anxiety in childhood and adolescence can cause serious impairment across many settings in a child's life. Childhood anxiety may be impacted by parent‐child attachment and by three specific parenting characteristics: acceptance, control and modelling of anxious behaviours. We review two evidence‐based family interventions for childhood and adolescent anxiety problems. Cognitive behavioural treatment is currently the most commonly used form of psychotherapy to treat specific anxieties. Global or diffuse anxiety, however, may be better suited to experiential interventions such as communications approaches to family treatment. Experiential therapists examine how dysfunctional family roles exacerbate childhood anxiety. These intervention techniques and their implications for social work practice were discussed.
Beneficial effects of iodized salt prophylaxis on thyroid volume in an iodine deficient area of southern Italy
Authors: Bonofiglio Daniela , Catalano Stefania , Perri Anna , Baldini Maria Pia , Marsico Stefania , Tagarelli Andrea , Conforti Domenico , Guido Rosita , Andò Sebastiano
Published: September 2008

Objective and subjects  Goitre prevalence in school‐age children is an indicator of the severity of iodine deficiency disorders (IDD) in an endemic area. The aims of the present study were (i) to provide ultrasound thyroid volume (TV) reference values in a healthy population of school‐children aged 11–14 year living in iodine‐sufficient areas of Calabria region (ii) to assess both goitre prevalence and urinary iodine (UI) concentration in all children aged 11–14 year from four mildly iodine‐deficient areas in which we have carried out a program of salt iodization and (iii) to evaluate the efficacy of the iodoprophylaxis in an adult population living in a small village of the same endemic area.

Design  Cross‐sectional and prospective studies.

Methods  TV was assessed by ultrasonography and iodine intake was estimated by measuring iodine excretion in spot urine samples.

Results  We provided the ultrasound normal reference values as a function of age and body surface area, which displayed significant differences from those recommended by the World Health Organization. By adopting local criteria, the prevalence of goitre in children ranged from 23·4% to 27·7% normalized for age and body surface area, respectively, while the UI excretion was < 100 µg/l in 38% of subjects studied. In an adult population living in the same endemic area, goitre prevalence was lowest in the 18–27‐year‐old age group, and increased progressively with age.

Conclusion  We propose for the first time local reference ultrasound values for TV in a population of 11–14‐year‐old school‐children that should be used for monitoring IDDs and have demonstrated the beneficial effects of iodoprophylaxis in consistent with reduced goitre prevalence in children and in the young adult population studied.
Collembola foraging responses to interacting fungi
Authors: ROTHERAY TIMOTHY D. , BODDY LYNNE , JONES T. HEFIN
Published: September 2008

Abstract1. Temperate woodland Collembola feed predominantly on plant detritus and fungi. They exhibit preferences for different fungi reflecting resource value in terms of Collembola growth and reproduction.

2. Studies of fungal grazing by Collembola have focused on non‐interacting fungi and have used one genetic isolate as representative of an entire fungal species. This study examines the effects of fungal interactions on Collembola behaviour, and elucidates differences arising from the use of genetically distinct fungal isolates.

3. Folsomia candidawere added to microcosms in which paired combinations of the fungi,Hypholoma fasciculare(four isolates),Phallus impudicus,Phanerochetae velutinaandResinicium bicolor(two isolates), were interacting. Collembola movement and mortality was recorded for 26 days. A constant preference for one fungal mycelium over the other was shown by Collembola in 12 of the 36 interactions studied. Lowest Collembola mortality was normally recorded on the preferred mycelium.

4. In 11 of the remaining 24 interactions, Collembola preference switched from one mycelium to the other; the greatest number of dead Collembola was recorded on the final preference.

5. Collembola behaviour on different fungal isolates was broadly similar, although in one species’ combination a change in fungal isolate resulted in a total reversal of Collembola preference. The implications of these results for decomposer food webs are considered.
TLR‐ and CXCR1‐dependent innate immunity: insights into the genetics of urinary tract infections
Authors: Ragnarsdóttir B. , Fischer H. , Godaly G. , Grönberg‐Hernandez J. , Gustafsson M. , Karpman D. , Lundstedt A. C. , Lutay N. , Rämisch S. , Svensson M. L. , Wullt B. , Yadav M. , Svanborg C.
Published: September 2008

The susceptibility to urinary tract infection (UTI) is controlled by the innate immune response and Toll like receptors (TLRs) are the sentinels of this response. If productive, TLR4 signalling may initiate the symptomatic disease process. In the absence of TLR4 signalling the infected host instead develops an asymptomatic carrier state. The activation of mucosal TLR4 is also influenced by the properties of the infecting strain, and pathogens use their virulence factors to trigger ‘pathogen‐specific’ TLR4 responses in the urinary tract but do not respond to the asymptomatic carrier strains in patients with asymptomatic bacteriuria (ABU). The TLR4 dependence has been demonstrated in mice and the relevance of low TLR4 function for protection for human disease was recently confirmed in children with asymptomatic bacteriuria, who expressed less TLR4 than age matched controls. Functional chemokines and functional chemokine receptors are crucial for neutrophil recruitment, and for the neutrophil dependent bacterial clearance. Interleukin (IL)‐8 receptor deficient mice develop acute septic infections and chronic tissue damage, due to aberrant neutrophil function. This mechanism is relevant for human UTI as pyelonephritis prone children express low levels of the human CXCL8 (Il‐8) receptor, CXC chemokine receptor 1 (CXCR1) and often have heterozygous CXCR1 polymorphisms.

This review illustrates how intimately the innate response and the susceptibility to UTI are linked and sophisticated recognition mechanisms that rely on microbial virulence and on host TLR4 and CXCR1 signalling.
Therapeutic challenges of urosepsis
Authors: Wagenlehner F. M. E. , Pilatz A. , Naber K. G. , Weidner W.
Published: September 2008

Urosepsis accounts for approximately 25% of all sepsis cases and may develop from a community or nosocomial acquired urinary tract infection (UTI). The underlying UTI is almost exclusively a complicated one with involvement of parenchymatous urogenital organs (e.g. kidneys, prostate). In urosepsis, as in other types of sepsis, the severity of sepsis depends mostly upon the host response. The treatment of urosepsis comprises four major aspects: Early goal directed therapy, early optimal pharmacodynamic exposure to antimicrobials, early control of the complicating factor in the urinary tract and specific sepsis therapy. Following these prerequisites there appear two major challenges that need to be addressed:

Firstly, time from admission to therapy is critical; the shorter the time to effective treatment, the higher the success rate. This aspect has to become incorporated into the organisational process. Secondly, adequate initial antibiotic therapy has to be insured. This goal implies however, a wide array of measures to ensure rational antibiotic policy. Both challenges are best targeted if an interdisciplinary approach at any level of the process is established, encompassing urologists, intensive care specialists, radiologists, microbiologists and clinical pharmacologists working tightly together at any time.
Asymptomatic bacteriuria and symptomatic urinary tract infections in pregnancy
Authors: Schnarr J. , Smaill F.
Published: September 2008

Symptomatic and asymptomatic bacteriuria is common in pregnant women. A history of previous urinary tract infections and low socioeconomic status are risk factors for bacteriuria in pregnancy. Escherichia coli is the most common aetiologic agent in both symptomatic and asymptomatic infection and quantitative culture is the gold standard for diagnosis. Treatment of asymptomatic bacteriuria has been shown to reduce the rate of pyelonephritis in pregnancy and therefore screening for and treatment of asymptomatic bacteriuria has become a standard of obstetrical care. Antibiotic treatment of asymptomatic bacteriuria is associated with a decrease in the incidence of low birth weight, but the methodological quality of the studies limits the strength of the conclusions that can be drawn. Debate exists in the literature as to whether treated pyelonephritis is associated with adverse fetal outcomes. There is no clear consensus in the literature on antibiotic choice or duration of therapy for infection. With increasing antibiotic resistance, consideration of local resistance rates is necessary when choosing therapy.
Experimental evidence of the dynamic effect of free‐floating plants on phytoplankton ecology
Authors: O’FARRELL INÉS , DE TEZANOS PINTO PAULA , RODRÍGUEZ PATRICIA L. , CHAPARRO GRISELDA , PIZARRO HAYDÉE N.
Published: September 2008

1. There is an increasing concern to understand the role of free‐floating plants (FFP) on the functioning and structure of shallow lakes, particularly the mechanisms by which their dominance is self‐stabilizing and how they may outcompete phytoplankton.

2. In a field experiment with mesocosms, we simulated three commonly encountered scenarios in warm temperate shallow lakes: FFP dominance, FFP fluctuation and FFP absence. We explored the effects of several key processes, triggered by FFP dynamics, on the composition, diversity and production of phytoplankton, and on the physicochemical conditions.

3. The effects of persistent floating mats on light interception triggered a complex response by the phytoplankton: species were lost and biomass was low, yet high diversity and productivity potential were maintained. A contrasting phytoplankton response characterized mesocosms lacking FFP, where light was sufficient but nitrogen was limiting. Fluctuating FFP cover brought periodic shifts between these two limiting resources for the phytoplankton, which most probably explain the maintenance of richness, diversity and production in these shallow lakes.

4. These results support the recently proposed framework of alternative stable states, providing experimental field evidence of the mechanisms resulting from the shifts between floating plants and phytoplankton dominance.
Involvement of the MP1 scaffold protein in ERK signaling regulation during Drosophila wing development
Authors: Mouchel‐Vielh Emmanuèle , Bloyer Sébastien , Salvaing Juliette , Randsholt Neel B. , Peronnet Frédérique
Published: September 2008

Mitogen‐activated protein kinase (MAPK) cascades are evolutionary conserved transduction pathways involved in many cellular processes. Kinase modules are associated with scaffold proteins that regulate signaling by providing critical spatial and temporal specificities. Some of these scaffold proteins have been shown to be conserved, both in sequence and function. In mouse, the scaffold MP1 (MEK Partner 1) forms a signaling complex with MEK1 and ERK1. In this work, we focus on Drosophila MP1 (dMP1). We show that dMP1 is expressed ubiquitously during embryonic and larval development. By in vitro and in vivo experiments, we show that dMP1 is located in the cytoplasm and the nuclei, and that it interacts with MEK and ERK. Genetic studies with transgenic Drosophila lines allowing either dMP1 over‐expression or dMP1 down‐regulation by RNA interference highlight dMP1 function in the control of cell differentiation during development of the Drosophila wing.
Stromal cells promote bone invasion by suppressing bone formation in ameloblastoma
Authors: Sathi G S A , Nagatsuka H , Tamamura R , Fujii M , Gunduz M , Inoue M , Rivera R S , Nagai N
Published: September 2008

Aims:  To study the stromal variation and role of stromal–tumour cell interaction in impaired bone formation as well as enhanced bone resorption in ameloblastoma.

Methods and results:  Four types of stroma were observed histologically; fibrous, desmoplastic, myxoid and myxoid with hyalinization. Osteoblast and osteoclast were counted using haematoxylin and eosin sections and immunohistochemistry with CD68. After histomorphometric analysis, only fibrous and myxoid types of stroma were distinctly identified. Secreted frizzled‐related peptide (sFRP)‐2, transforming growth factor‐beta 1 and receptor activator of nuclear factor‐κB ligand (RANKL) revealed strong expression in myxoid type compared with the normal stroma. Bone morphogenetic protein (BMP)‐2 was negative in myxoid type, but positive in normal stroma. Fibrous‐type stroma showed weak expression of all antigens except RANKL compared with myxoid type.

Conclusions:  The results suggest that stroma does not act only in bone resorption, but also in the suppression of new bone formation. sFRP‐2 is the main factor for impaired bone formation. The expression of markers related to osteoclastogenesis and suppression of osteoblast formation is higher in myxoid‐type than in fibrous‐type stroma. Tumour cells create a favourable environment for impaired bone formation by secreting sFRP‐2 as well as bone resorption by secreting RANKL and interleukin‐6.
Capillary apposition and density in the diagnosis of alveolar capillary dysplasia
Authors: Melly L , Sebire N J , Malone M , Nicholson A G
Published: September 2008

Aim:  Alveolar capillary dysplasia (ACD) is a rare disorder, typically presenting with persistent pulmonary hypertension of the newborn. The aim was to characterise further the histological features of patients suspected of having ACD and to correlate histopathological features with outcome.

Methods and results:  Three pathologists retrospectively reviewed 21 surgical lung biopsy specimens (SLBx) where ACD entered the differential diagnosis. Semi‐quantitative assessment showed that there was a spectrum of muscular arterial hypertrophy, capillary apposition to epithelium and capillary density within the interstitium, with the latter being more disordered in ACD. Misalignment of pulmonary vessels was also frequently seen. Four of 19 patients survived beyond the neonatal period, these having higher degrees of capillary apposition and density. Associated extrapulmonary abnormalities were common, most frequently with ACD.

Conclusion:  Poor capillary apposition and density, allied with medial arterial hypertrophy and misalignment of pulmonary vessels are the strongest diagnostic features of ACD. Of the four patients alive, all had high capillary apposition and density, suggesting that these features may be of prognostic value. SLBx remains useful in such cases as it may help predict patients who survive the neonatal period and also identify patients with disorders that are not primarily vascular anomalies.
Salivary duct carcinoma in situ of the parotid gland
Authors: Simpson R H W , Desai S , Di Palma S
Published: September 2008

Aims:  To describe three cases of purely in situ salivary duct carcinoma, so as better to define the entity.

Methods and results:  Three primary tumours of the parotid gland are presented, in each case composed of cysts and ducts and lined by high nuclear grade epithelial cells. All parts of each tumour were surrounded by a myoepithelial cell rim and there was no evidence of invasion. The tumour cells expressed immunohistochemical markers seen in invasive salivary duct carcinoma of usual (high‐grade) type. In two cases the androgen receptor (AR) reaction was strong, but there was no immunohistochemical expression of HER2 protein or gene amplification by in situ hybridization. In the remaining case, fewer nuclei stained for AR, but both HER2 protein and gene amplification were demonstrated.

Conclusions:  Salivary duct carcinoma in situ is morphologically similar to breast ductal carcinoma in situ and, although our cases are few, salivary duct carcinoma in situ can possibly be subdivided into luminal and non‐luminal cell types, as can analogous mammary neoplasms. The present study cannot determine whether low‐grade cribriform cystadenocarcinoma, architecturally similar but immunohistochemically different, is part of the spectrum of salivary duct carcinoma in situ, or whether it represents a separate entity.
Study of matrix metalloproteinases and their tissue inhibitors in ductal in situ carcinomas of the breast
Authors: Gonzalez L O , Corte M D , Vazquez J , Junquera S , Sanchez R , Viña A , Rodriguez J C , Lamelas M L , Vizoso F
Published: September 2008

Aims:  To analyse the expression of metalloproteinases (MMPs) and their inhibitors (TIMPs) in ductal carcinoma in situ of the breast (DCIS).

Methods and results:  An immunohistochemical study was performed in 56 patients with pure DCIS, in 39 with DCIS adjacent to invasive carcinoma (IDC) and 63 patients with T1 IDC, using tissue microarrays and specific antibodies against MMPs and TIMPs. Immunohistochemical results were categorized using a specific software program. The data were analysed by unsupervised hierarchical cluster analysis by each cellular type. IDC showed a higher expression rate of MMP‐7 and TIMP‐1 than pure DCIS, as well as a higher expression rate of MMP‐9 and TIMP‐3 than the DCIS component of mixed cases, whereas pure DCIS showed a higher rate of expression of MMP‐9 and ‐11 and TIMP‐3 than in the DCIS component of mixed cases. Pure DCIS with a periductal inflammatory infiltrate showed significantly higher MMP‐2, ‐14 and TIMP‐1. Dendograms identified two cluster groups with distinct MMP/TIMP expression profiles in neoplastic cells and fibroblastic or mononuclear inflammatory cells surrounding the neoplastic ducts of pure DCIS.

Conclusions:  The results indicate the distinct variability in MMP/TIMP expression by DCIS, which may be of potential biological and clinical interest in breast cancer.
Activation of the NF‐κB signalling pathway in diffuse large B‐cell lymphoma: clinical implications
Authors: Espinosa I , Briones J , Bordes R , Brunet S , Martino R , Sureda A , Sierra J , Prat J
Published: September 2008

Aim:  To characterize the activation of the nuclear factor (NF)‐κB pathway in diffuse large B‐cell lymphoma (DLBCL) by immunohistochemistry.

Methods and results:  Sixty‐six DLBCLs treated with anthracycline‐containing chemotherapy were evaluated with antibodies against phosphorylated p65 (P‐p65), p65, p50, p52, IKKα, and phosphorylated IκB (P‐IκB). NF‐κB activation was based on the expression of P‐p65, P‐IκB, and nuclear expression of p65 or p52 in the tumour cells. P‐p65 and P‐IκB were expressed in 13 (20%) and 17 cases (26%), respectively. p65, p52 and IKKα were found in the cytoplasm. A correlation was found between expression of P‐p65 and P‐IκB (P < 0.0001), but not between the two subtypes of DLBCL [germinal centre B cell and non‐germinal centre (GC)]. P‐p65+ tumours showed a better response to chemotherapy (P = 0.025) and a trend to increased event‐free survival (P = 0.08). However, P‐IκB expression was not associated with either clinical response or survival. Bcl‐2 was not preferentially expressed on DLBCL tumours with NF‐κB activation, as determined by expression of P‐p65 and P‐IκB proteins.

Conclusions:  NF‐κB activation in DLBCL is preferentially mediated through the classical pathway and a novel mechanism involving phosphorylation of p65. Activation of NF‐κB by P‐p65 is associated with good prognosis. NF‐κB activation is not confined to non‐GC DLBCL exclusively.
Individual and combined responses of stream ecosystems to multiple stressors
Authors: Townsend Colin R. , Uhlmann S. Sebastian , Matthaei Christoph D.
Published: September 2008

Managers must understand the effects of stressors on ecosystems in order to identify thresholds of harm but, to be meaningful, thresholds will usually need to be defined for situations where multiple stressors are operating.

We investigated the individual and combined effects of the principal stressors (nutrient concentration and streambed fine sediment cover) operating in native grassland streams converted to pasture in New Zealand, using two different approaches: a survey of 32 small streams and an experiment involving nine streams where the stressors were manipulated in a factorial design. We investigated the consequences for populations of benthic invertebrates and for the structure of communities, including taxon richness and the representation of species traits.

Up to half the taxa and most community metrics responded to at least one stressor. Our results suggest that in these streams, an increase in fine sediment loading from anthropogenic causes had more widespread effects than augmented nutrient concentrations. Of most significance is our finding, both from the survey and, in particular, the experiment, of a variety of complex interactions among the stressors.

Synthesis and applications. The development of indices of stream health that distinguish the effects of sediment from those of nutrients should help prioritize catchment management actions. Of more general importance is our finding that the consequences of stressors are often unpredictable on the basis of knowledge of single effects; if managers only consider the effects of individual stressors, their assessment of risk may be higher or lower than reality.
Impacts of massive landscape change on a carnivorous marsupial in south‐eastern Australia: inferences from landscape genetics analysis
Authors: Lada Hania , Thomson James R. , Mac Nally Ralph , Taylor Andrea C.
Published: September 2008

In much of the world, fauna has been adversely affected by human actions, including conversion of forests to farmland, logging and regulation of river flows. Landscape genetics data can provide information about dispersal and gene flow across the landscape, identifying barriers and facilitators of gene flow. Landscapes of central Victoria, Australia, have been altered extensively in the last 160 years. Much vegetation has been cleared or degraded, and only forest patches of mainly re‐growth remain, yet some forest‐dependent species like the yellow‐footed antechinus Antechinus flavipes persist. The antechinus has good dispersal capabilities and is the only native, small, carnivorous mammal on most floodplains. We use antechinus as a model to understand species persistence in fragmented landscapes.

We analysed variation at 11 microsatellite loci and the control region of mitochondrial DNA to infer past and contemporary gene flow among A. flavipes populations. To explore genetic connectivity, we used least‐cost path methods, which assign different ‘friction’ costs to vegetation, cleared land, roads and rivers.

Populations from 11 forests formed six distinct genetic groups, and with few exceptions, animals from nearby forests clustered together despite the intervening Murray River or farmland with only narrow vegetation corridors between them.

Genetic connectivity was aided by corridors of vegetation and inhibited by cleared land.

Synthesis and applications. Our approach, capitalizing on inferences on both historic and contemporary gene flow, provides management agencies with key information on metapopulation dynamics in landscapes. Rather than merely maintaining existing vegetation upon which this (and many other) species depend, the genetic information also informs where future plantings should be prioritized to facilitate demographic and genetic exchange among sub‐populations of species. Moreover, the decline in condition (‘health’) of riparian trees in this region must be reversed by provision of flooding flows; otherwise, metapopulation dynamics will become even more disarticulated than at present.
Response of broadleaved evergreen Mediterranean forest vegetation to fire disturbance during the Holocene: insights from the peri‐Adriatic region
Authors: Colombaroli Daniele , Tinner Willy , Van Leeuwen Jacqueline , Noti Roland , Vescovi Elisa , Vannière Boris , Magny Michel , Schmidt Roland , Bugmann Harald
Published: September 2008

Aim  To test whether fire contributed to the expansion and compositional change of evergreen forests in the Mediterranean region during the Holocene.

Location  The peri‐Adriatic region, encompassing the Italian peninsula, Sicily and the western and southern Balkans between latitudes 46° and 37° N.

Methods  New high‐resolution pollen and microscopic charcoal data from Lago dell’Accesa (Tuscany, Italy) were used to estimate the response of the evergreen oak, Quercus ilex L., to fire during its expansion phase at 8500 cal. yr bp. The data were compared with the pollen and charcoal series from other Mediterranean sites (Lago di Massaciuccoli in Tuscany, Malo Jezero in Croatia, Biviere di Gela in Sicily) and analysed using numerical techniques (redundancy analysis, detrended canonical correspondence analysis) to identify long‐term fire–vegetation linkages and the degree of compositional change.

Results  Microscopic charcoal and pollen of evergreen oaks were negatively correlated during the period of quasi‐natural fire regime (Mesolithic, 10,000–8000 cal. yr bp). In addition, there was no such positive correlation during periods when the fire regime was potentially more influenced by people (Neolithic–Bronze Age, 8000–3000 cal. yr bp). Compared with inland sites, coastal sites that are currently located at a distinct ecotone showed more compositional change.

Main conclusions  The analyses suggest that climatic change, without an additional effect of fire regimes, favoured the expansion and compositional change of evergreen forests across the peri‐Adriatic region. Strikingly different patterns occurred along a north–south gradient. In the north (Tuscany and Croatia, meso‐Mediterranean belt), Q. ilex replaced deciduous forests when conditions became drier; in the south (Sicily, thermo‐Mediterranean belt) the species replaced maquis or steppe vegetation when climatic conditions became moister. We conclude that the projected increase in fire activity may lead to the loss of most of the remaining relict forests of Q. ilex in southern Europe.
Y‐chromosome analysis confirms highly sex‐biased dispersal and suggests a low male effective population size in bonobos (Pan paniscus)
Authors: ERIKSSON JONAS , SIEDEL HEIKE , LUKAS DIETER , KAYSER MANFRED , ERLER AXEL , HASHIMOTO CHIE , HOHMANN GOTTFRIED , BOESCH CHRISTOPHE , VIGILANT LINDA
Published: September 2008

Dispersal is a rare event that is difficult to observe in slowly maturing, long‐lived wild animal species such as the bonobo. In this study we used sex‐linked (mitochondrial DNA sequence and Y‐chromosome microsatellite) markers from the same set of individuals to estimate the magnitude of difference in effective dispersal between the sexes and to investigate the long‐term demographic history of bonobos. We sampled 34 males from four distinct geographical areas across the bonobo distribution range. As predicted for a female‐dispersing species, we found much higher levels of differentiation among local bonobo populations based upon Y‐chromosomal than mtDNA genetic variation. Specifically, almost all of the Y‐chromosomal variation distinguished populations, while nearly all of the mtDNA variation was shared between populations. Furthermore, genetic distance correlated with geographical distance for mtDNA but not for the Y chromosome. Female bonobos have a much higher migration rate and/or effective population size as compared to males, and the estimate for the mitochondrial TMRCA (time to most recent common ancestor) was approximately 10 times greater than the estimate for the Y chromosome (410 000 vs. 40 000–45 000). For humans the difference is merely a factor of two, suggesting a more stable demographic history in bonobos in comparison to humans.
Fitness of hybrids between rapeseed (Brassica napus) and wild Brassica rapa in natural habitats
Authors: ALLAINGUILLAUME J. , ALEXANDER M. , BULLOCK J. M. , SAUNDERS M. , ALLENDER C. J. , KING G. , FORD C. S. , WILKINSON M. J.
Published: September 2008

Fitness of hybrids between genetically modified (GM) crops and wild relatives influences the likelihood of ecological harm. We measured fitness components in spontaneous (non‐GM) rapeseed × Brassica rapa hybrids in natural populations. The F1 hybrids yielded 46.9% seed output of B. rapa, were 16.9% as effective as males on B. rapa and exhibited increased self‐pollination. Assuming 100% GM rapeseed cultivation, we conservatively predict < 7000 second‐generation transgenic hybrids annually in the United Kingdom (i.e. ∼20% of F1 hybrids). Conversely, whilst reduced hybrid fitness improves feasibility of bio‐containment, stage projection matrices suggests broad scope for some transgenes to offset this effect by enhancing fitness.
History vs. current demography: explaining the genetic population structure of the common frog (Rana temporaria)
Authors: JOHANSSON MARKUS , PRIMMER CRAIG R. , MERILÄ JUHA
Published: September 2008

The amount of genetic variability at neutral marker loci is expected to decrease, and the degree of genetic differentiation among populations to increase, as a negative function of effective population size. We assessed the patterns of genetic variability and differentiation at seven microsatellite loci in the common frog (Rana temporaria) in a hierarchical sampling scheme involving three regions (208–885 km apart), three subregions within regions and nine populations (5–20 km apart) within subregions, and related the variability and differentiation estimates to variation in local population size estimates. Genetic variability within local populations decreased significantly with increasing latitude, as well as with decreasing population size and regional site occupancy (proportion of censured localities occupied). The positive relationship between population size and genetic variability estimates was evident also when the effect of latitude (cf. colonization history) was accounted for. Significant genetic differentiation was found at all hierarchical levels, and the degree of population differentiation tended to increase with increasing latitude. Isolation by distance was evident especially at the regional sampling level, and its strength increased significantly towards the north in concordance with decreasing census and marker‐based neighbourhood size estimates. These results are in line with the conjecture that the influence of current demographic factors can override the influence of historical factors on species population genetic structure. Further, the observed reductions in genetic variability and increased degree of population differentiation towards the north are in line with theoretical and empirical treatments suggesting that effective population sizes decline towards the periphery of a species’ range.
The enigmatic invasiveSpartina densiflora: A history of hybridizations in a polyploidy context
Authors: FORTUNE P. M. , SCHIERENBECK K. , AYRES D. , BORTOLUS A. , CATRICE O. , BROWN S. , AINOUCHE M. L.
Published: September 2008

The aim of this study was to explore the origin of the invasiveSpartina densifloraby analysing samples from the native region (South America) and from a recently colonized area (California). A combination of various molecular data (chloroplast and nuclear sequences, molecular fingerprint) and ploidy level estimations was used to answer the question whether the reticulate phylogenetic pattern previously detected inS. densiflorawas restricted to California, or alternatively, whether a more ancient hybrid origin preceded formation of this species in its native area. We found that this species is heptaploid in both its native and introduced range. Identification of nuclear homeologous sequences indicate that this species has a reticulate origin in its native range, involving a lineage related to the hexaploid clade formed byS. alterniflora,S. foliosa, andS. maritima, and another lineage related to the sub‐Antarctic endemicS. arundinaceathat provided the chloroplast genome. The samples from California displayed similar multilocus patterns to the samples from Chile, supporting the hypothesis that this species originated on the southeast American coast (Argentina), from where it eventually spread to the west coast of South America (Chile) first and to the Northern Hemisphere (California) later.
Interdependence of two NarK domains in a fused nitrate/nitrite transporter
Authors: Goddard Alan D. , Moir James W. B. , Richardson David J. , Ferguson Stuart J.
Published: September 2008

Nitrate uptake is essential for various bacterial processes and combines with nitrite export to form the usual initial steps of denitrification, a process that reduces nitrate to dinitrogen gas. Although many bacterial species contain NarK‐like transporters that are proposed to function as either nitrate/proton symporters or nitrate/nitrite antiporters based on sequence homology, these transporters remain, in general, poorly characterized. Several bacteria appear to contain a transporter that is a fusion of two NarK‐like proteins, although the significance of this arrangement remains elusive. We demonstrate that NarK fromParacoccus denitrificansis expressed as a fusion of two NarK‐like transporters. NarK1 and NarK2 are separately capable of supporting anaerobic denitrifying growth but with growth defects that are partially mitigated by coexpression of the two domains. NarK1 appears to be a nitrate/proton symporter with high affinity for nitrate and NarK2 a nitrate/nitrite antiporter with lower affinity for nitrate. Each transporter requires two conserved arginine residues for activity. A transporter consisting of inactivated NarK1 fused to active NarK2 has a dramatically increased affinity for nitrate compared with NarK2 alone, implying a functional interaction between the two domains. A potential model for nitrate and nitrite transport inP. denitrificansis proposed.
Endogenous and exogenous ghrelin enhance the colonic and gastric manifestations of dextran sodium sulphate‐induced colitis in mice
Authors: De Smet B. , Thijs T. , Moechars D. , Colsoul B. , Polders L. , Ver Donck L. , Coulie B. , Peeters T. L. , Depoortere I.
Published: September 2008

Abstract  Ghrelin is an important orexigenic peptide that not only exerts gastroprokinetic but also immunoregulatory effects. This study aimed to assess the role of endogenous and exogenous ghrelin in the pathogenesis of colitis and in the disturbances of gastric emptying and colonic contractility during this process. Dextran sodium sulphate colitis was induced for 5 days in (i) ghrelin+/+ and ghrelin−/− mice and clinical and histological parameters were monitored at days 5, 10 and 26 and (ii) in Naval Medical Research Institute non‐inbred Swiss (NMRI) mice treated with ghrelin (100 nmol kg−1) twice daily for 5 or 10 days. Neural contractility changes were measured in colonic smooth muscle strips, whereas gastric emptying was measured with the 14C octanoic acid breath test. Inflammation increased ghrelin plasma levels. Body weight loss, histological damage, myeloperoxidase activity and IL‐1β levels were attenuated in ghrelin−/− mice. Whereas absence of ghrelin did not affect changes in colonic contractility, gastric emptying in the acute phase was accelerated in ghrelin+/+ but not in ghrelin−/− mice. In agreement with the studies in ghrelin knockout mice, 10 days treatment of NMRI mice with exogenous ghrelin enhanced the clinical disease activity and promoted infiltration of neutrophils and colonic IL‐1β levels. Unexpectedly, ghrelin treatment decreased excitatory and inhibitory neural responses in the colon of healthy but not of inflamed NMRI mice. Endogenous ghrelin enhances the course of the inflammatory process and is involved in the disturbances of gastric emptying associated with colitis. Treatment with exogenous ghrelin aggravates colitis, thereby limiting the potential therapeutic properties of ghrelin during intestinal inflammation.
Effect of a κ‐opioid agonist, i.v. JNJ‐38488502, on sensation of colonic distensions in healthy male volunteers
Authors: Floyd B. N. , Camilleri M. , Busciglio I. , Sweetser S. , Burton D. , Wong G. Y. , Kell S. , Khanna S. , Hwang S. , Zinsmeister A. R.
Published: September 2008

Abstract Kappa‐opioid receptors are located on visceral pain fibres. JNJ‐38488502 is a highly selective tetrapeptide κ‐opioid agonist with little access to the central nervous system and low risk of central nervous system side effects. The aim of the study was to evaluate the effects of i.v. JNJ‐38488502 on sensations, including pain, during colonic distension. In a single‐centre study, 23 healthy adult males underwent a single‐dose, randomized, double‐blind crossover study of JNJ‐38488502 (0.42 mg kg−1i.v. infusion)vsplacebo on left colon compliance, sensory thresholds and ratings during standard distensions. One participant could not undergo sensation studies. In the other 22, JNJ‐38488502 increased colonic compliance (pressure at half‐maximum volume 17.9 ± 0.8 mmHg) compared to placebo (21.6 ± 0.9 mmHg,P = 0.007). There was no significant effect on sensory thresholds which, however, were not reached by 44 mmHg in >50% of participants in both treatment phases. There were no significant treatment effects on sensory ratings to distensions at 8, 16, 24, 32 and 36 mmHg above baseline operating pressure. JNJ‐38488502 was associated with increased urine output and plasma prolactin, consistent with κ‐opioid receptor activation. This study concluded that i.v. JNJ‐38488502 induced κ‐opioid effects, but did not attenuate colonic sensations following random order colonic distension. Further studies of effects on pain sensations in health and disease are required.
CD8+CD60+ T Cells, Cells Expressing Epsilon Specific mRNA and Th1/Th2 Cytokines in Cord Blood and at 7 Months of Age
Authors: Smith‐Norowitz T. A. , Norowitz K. B. , Silverberg J. I. , Chice S. , Shah V. , Stanek A. , Walker D. , Brennan J. P. , Durkin H. G. , Bluth M. H.
Published: September 2008

IgE levels in cord blood have been investigated as predictors of atopy, but no definitive findings have been made. Other factors, including cells and/or cytokines may serve as predictors of this disease. Cord blood and peripheral blood was obtained at birth and at 7 months of age, respectively, from children (n = 2) with a family history of allergy. Cells in cord blood and peripheral blood were phenotyped and levels of serum immunoglobulins (IgM, IgG, IgA and IgE) were determined. In addition, placentas from these pregnancies were obtained and stained for IgE+ cells and CD8+CD60+ T cells. We found immunoglobulin levels were within normal ranges although IgE levels were negligible in cord blood and at 7 months of age. Similar numbers of CD8+ T cells and CD19+ B cells were detected in cord blood and at 7 months of age. However, CD4+ T cells increased (twofold) and CD16+/CD56+ natural killer precursor cells decreased (twofold) at 7 months of age. CD8+ T cells in their cord blood and at 7 months of age comprised of >50% CD8+CD60+ T cells. Cord blood cells expressed epsilon‐specific mRNA and mRNA for interleukin‐2 (IL‐2), IL‐4, IL‐10 and interferon‐γ (IFN‐γ) but not IL‐6. At 7 months of age, peripheral blood mononuclear cells expressed epsilon‐specific mRNA and mRNA for all cytokines. In the placental membrane, we detected IgE+ cells, while CD8+CD60+ T cells were detected in the chorionic villi. CD8+CD60+ T cells, cells expressing epsilon‐specific and IL‐6‐specific mRNA may contribute to the pathobiology and provide important prognostic indicators of atopy.
Differential Invasion of Trypanosoma brucei brucei and Lymphocytes into the Brain of C57BL/6 and 129Sv/Ev Mice
Authors: Masocha W. , Amin D. N. , Kristensson K. , Rottenberg M. E.
Published: September 2008

Trypanosoma brucei subspecies invade the brain parenchyma at late stages of human and experimental rodent infections. In this study, we compared the outcome of infection with T. b. brucei in MHC‐matched (H‐2b) C57BL/6 (B6) and 129Sv/Ev (Sv‐129). Sv‐129 showed higher parasitaemia and lower specific IgM (but not IgG) antibody levels than B6 mice. The number of trypanosomes, CD4+ and CD8+ T cells in the brain parenchyma was higher in B6 mice. B6 mice lost weight and showed higher cumulative mortality when compared with Sv‐129 mice. Higher levels of IL‐1β, IL‐6, IL‐10, TNF‐α, IFN‐γ, ICAM‐1 and E‐selectin, but low levels of TGF‐β mRNA were present in brains of B6 when compared with Sv‐129‐infected mice. Thus, host genetics differentially determine the invasion of T. b. brucei into the brain parenchyma, which is paralleled by the severity of inflammation in the brain and course of the disease, but not by parasitaemia nor by antibody titres.
Effect of Neonatal Sublingual Vaccination with Native or Denatured Ovalbumin and Adjuvant CpG or Cholera Toxin on Systemic and Mucosal Immunity in Mice
Authors: Huang C.‐F. , Wu T.‐C. , Chu Y.‐H. , Hwang K.‐S. , Wang C.‐C. , Peng H.‐J.
Published: September 2008

Sublingual immunotherapy has been applied for allergic diseases, but whether sublingual immunization in neonates can prevent sensitization has not been studied. In this study, we evaluate the effect of neonatal sublingual vaccination with native or denatured allergens alone or plus adjuvant on allergy prevention. Newborn BALB/c mice were sublingually vaccinated daily for the first 3 days with native or denatured ovalbumin (OVA) only, or combined adjuvant CpG or cholera toxin (CT). They were sensitized with OVA adsorbed onto alum 7 weeks after the last vaccination. Specific secretory IgA antibody responses were readily induced by neonatal vaccination with antigen plus CpG or CT, but not with antigen alone. Whereas vaccination with denatured OVA plus CpG markedly enhanced T helper 1 (Th1) responses and inhibited IgE production, vaccination with denatured OVA plus CT increased cervical lymph node cell production of interleukin‐4 (IL‐4), IL‐5, IL‐6, and serum IgG1 responses. These data demonstrate that neonatal sublingual vaccination with denatured OVA and CpG not only preferentially induces systemic Th1 responses and mucosal immunity, but also simultaneously abrogates IgE production. Neonatal sublingual vaccines may play a role for the strategy of allergy prevention.