bluuebunny/crossref_metadata_2025 huggingface dataset:
Habitat characteristics of the rare underground orchid Rhizanthella gardneri
Authors: Bougoure Jeremy , Brundrett Mark , Brown Andrew , Grierson Pauline F.
Published: September 2008

Rhizanthella gardneri R.S.Rogers is an entirely subterranean mycoheterotrophic orchid known only from two isolated populations within south-western Western Australia (WA). This rare species appears restricted to habitats dominated by species of the Melaleuca uncinata complex. R. gardneri purportedly forms a tripartite relationship with Melaleuca1, via a connecting mycorrhizal fungus, for the purpose of carbohydrate and nutrient acquisition. Here, we quantify key climate, soil and vegetation characteristics of known R. gardneri habitats to provide baseline data for monitoring of known R. gardneri populations, to better understand how R. gardneri interacts with its habitat and to identify possible new sites for R. gardneri introduction. We found that the habitats of the two known R. gardneri populations show considerable differences in soil chemistry, Melaleuca structure and Melaleuca productivity. Multivariate analyses showed that both multidimensional scaling (MDS) and principal components analysis (PCA) ordinations of soil chemical characteristics were very similar. Individual sites within populations were relatively similar in all attributes measured, whereas overall northern and southern habitats were distinct from each other. These results suggest that R. gardneri can tolerate a range of conditions and may be more widespread than previously thought, given that there are extensive areas of Melaleuca thickets with similar habitat characteristics across south-western WA. Variability within the habitats of known R. gardneri populations suggests translocation of this species into sites with similar vegetation may be a viable option for the survival of this species.
A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
Authors: Pereyra-Bonnet F. , Fernández-Martín R. , Olivera R. , Jarazo J. , Vichera G. , Gibbons A. , Salamone D.
Published: September 2008

Transgenesis is an essential tool in many biotechnological applications. Intracytoplasmic sperm injection (ICSI)-mediated gene transfer is a powerful technique to obtain transgenic pups; however, most domestic animal embryos do not develop properly after ICSI. An additional step in the protocol, namely assistance by haploid chemical activation, permits the use of ICSI-mediated gene transfer to generate transgenic preimplantation embryos in a wide range of domestic species, including ovine, porcine, feline, equine and bovine. In the present study, spermatozoa from five species were coincubated with pCX-EGFP plasmid and injected into metaphase II oocytes. The chemical activation protocol consisted of ionomycin plus 6-dimethylaminopurine. We detected high proportions of fluorescent EGFP embryos for all five species (23–60%), but with a high frequency of mosaic expression (range 60–85%). To our knowledge, this is the first study to produce exogenous DNA expression in feline and equine embryos. Chemical activation reduces the lag phase of egfp expression in ovine embryos. Our results show that this unique method could be used to obtain ovine, porcine, feline, bovine and equine transgenic preimplantation embryos.
Technical Note: Effect of density gradient centrifugation with trypsin on the in vivo fertilising capability of bovine spermatozoa
Authors: Blevins Brock A. , de la Rey Morne , Loskutoff Naida M.
Published: September 2008

The present study investigated the effect of a novel density gradient centrifugation (DGC) treatment using recombinant trypsin on the in vivo fertilising capability of bovine spermatozoa compared with a standard method. In Trial 1, semen collected from Boran and Ankole (Bos indicus) bulls was treated either with a silane-coated silica particle colloid formulated for humans with a recombinant trypsin or processed using a standard method (dilution in an egg yolk-based diluent). Semen processed by the two methods was used to artificially inseminate (AI) superovulated cattle. Day 7 embryos were flushed and assessed for fertilisation rates and embryo quality. Trial 2 used a trypsinised silane-coated silica particle colloid formulated specifically for bovine semen. Trial 1 resulted in significantly higher fertilisation rates using the trypsinised human DGC treatment than cows inseminated using the standard method (75.2% v. 67%, respectively; P < 0.01), but the numbers of transferable-quality Day 7 embryos did not differ between the two groups (P > 0.05). Results for Trial 2 indicated that cows inseminated with the trypsinised bovine DGC treatment had significantly increased fertilisation rates compared with the standard method (88.4% v. 63.1%, respectively; P < 0.01) and had significantly higher numbers of transferable-quality embryos (70.3% v. 51.8%, respectively; P < 0.01). In summary, bovine sperm treatment before AI by DGC and recombinant trypsin increases fertilisation rates and can result in more transferable-quality embryos compared with standard methods.
Temporal dynamics of ram sperm binding and survival during 48-h coculture with oviducal epithelial cells
Authors: Lloyd R. E. , Badia E. , Fazeli A. , Watson P. F. , Holt W. V.
Published: September 2008

Following insemination, ram spermatozoa bind to oviducal epithelial cells (OEC) in vivo and remain viable for several hours before fertilisation. In the present study, we investigated whether OEC monolayers reproduce this effect in vitro, performing an analysis of ram sperm binding and survival over an extended (48 h) period at 39°C. We wanted to determine whether the reproductive cycle phase and/or oviducal region would influence ram sperm binding and survival in coculture with OEC and whether reproductive and non-reproductive epithelial cells bound and maintained the viability of ram spermatozoa equivalently. Oviducts were separated into groups based on their ovarian state (follicular or luteal) and then divided into two parts (isthmus and ampulla) for OEC isolation. Sheep kidney epithelial cells (Madin-Darby ovine kidney; MDOK) were purchased commercially. Reproductive cycle phase, but not oviducal region, affected sperm binding to OEC. Although more spermatozoa bound to luteal OEC than to follicular OEC at 1 h, at 24 h follicular OEC had bound more spermatozoa than luteal OEC. Generally, spermatozoa that were bound to OEC and MDOK had enhanced viability at each of the time points investigated (1, 6, 24 and 48 h), but the viability of the OEC-bound spermatozoa was greater than that of the MDOK-bound spermatozoa at 48 h. In conclusion, ram sperm–epithelial cell interactions are temporal, dynamic and depend on the origin of the epithelial cells.
Semen collection, characterisation and artificial insemination in the beluga (Delphinapterus leucas) using liquid-stored spermatozoa
Authors: O'Brien J. K. , Steinman K. J. , Schmitt T. , Robeck T. R.
Published: September 2008

Ejaculates were collected from a beluga (Delphinapterus leucas) to gain an understanding of sperm biology and develop a short-term sperm preservation method for use in artificial insemination (AI). Ejaculate parameters and biochemistry, semen production and serum testosterone concentrations of an adult male were characterised for 21 months. Sperm viability, acrosome integrity and morphology did not change (P > 0.05) but ejaculate volume, sperm concentration and total spermatozoa per ejaculate were higher (P < 0.05) from January to June than from July to December. Peak testosterone concentrations (P < 0.05) were observed from October to April (8.0 ± 1.6 ng mL–1). The effects of hyaluronic acid (HA), antioxidants, storage temperature and time on in vitro sperm characteristics were examined. Motility parameters and viability were improved (P < 0.05) when semen was stored at 5°C compared with 21°C. During the first 24 h of storage sperm agglutination was absent only at 5°C in the presence of HA. A nulliparous 28-year-old female was inseminated endoscopically with liquid-stored semen. A pregnancy and birth of a calf was achieved following AI for the first time in this species, thereby validating both the AI technique and the fertility of beluga spermatozoa after chilled storage in a specialised diluent.
Expression of eukaryotic elongation initiation factor 1A differentially marks zygotic genome activation in biparental and parthenogenetic porcine embryos and correlates with in vitro developmental potential
Authors: Magnani Luca , Johnson Christine M. , Cabot Ryan A.
Published: September 2008

Zygotic genome activation (ZGA) is a major event during cleavage development. In vitro manipulation of mammalian embryos (including embryo culture) can result in developmental arrest around the time of ZGA. Eukaryotic elongation initiation factor 1A (eIF1A) has been used as a marker for ZGA in some mammalian species. We hypothesised expression of eIF1A can be used to assess ZGA in the pig; we also hypothesised that the expression profile of eIF1A can be used to assess developmental potential in vitro. The aims of the present study were to determine the expression pattern of eIF1A during porcine cleavage development and to assess its expression levels in embryos of different quality. We used a real-time reverse transcription–polymerase chain reaction assay to quantify eIF1A transcripts at different time points during cleavage development in porcine embryos produced by parthenogenetic activation (PA) and in vitro fertilisation (IVF). We found that eIF1A is activated at the two-cell stage in IVF embryos and at the four-cell stage in PA embryos. We showed that the increase in transcript levels observed in parthenogenetic embryos is dependent on de novo transcription. We found altered levels of eIF1A transcripts in parthenogenetic embryos that presented as either two- or eight-cell embryos 48 h after activation compared with four-cell embryos at the same time point. Our work supports the hypothesis that eIF1A is a marker of porcine ZGA and its expression profile can be used to assess embryo quality.
Molecular control of mitochondrial function in developing rhesus monkey oocytes and preimplantation-stage embryos
Authors: Mtango N. R. , Harvey A. J. , Latham K. E. , Brenner C. A.
Published: September 2008

The mitochondrion undergoes significant functional and structural changes, as well as an increase in number, during preimplantation embryonic development. The mitochondrion generates ATP and regulates a range of cellular processes, such as signal transduction and apoptosis. Therefore, mitochondria contribute to overall oocyte quality and embryo developmental competence. The present study identified, for the first time, the detailed temporal expression of mRNAs related to mitochondrial biogenesis in rhesus monkey oocytes and embryos. Persistent expression of maternally encoded mRNAs was observed, in combination with transcriptional activation and mRNA accumulation at the eight-cell stage, around the time of embryonic genome activation. The expression of these transcripts was significantly altered in oocytes and embryos with reduced developmental potential. In these embryos, most maternally encoded transcripts were precociously depleted. Embryo culture and specific culture media affected the expression of some of these transcripts, including a deficiency in the expression of key transcriptional regulators. Several genes involved in regulating mitochondrial transcription and replication are similarly affected by in vitro conditions and their downregulation may be instrumental in maintaining the mRNA profiles of mitochondrially encoded genes observed in the present study. These data support the hypothesis that the molecular control of mitochondrial biogenesis, and therefore mitochondrial function, is impaired in in vitro-cultured embryos. These results highlight the need for additional studies in human and non-human primate model species to determine how mitochondrial biogenesis can be altered by oocyte and embryo manipulation protocols and whether this affects physiological function in progeny.
Glycosidase determination in bovine oviducal fluid at the follicular and luteal phases of the oestrous cycle
Authors: Carrasco Luis Cesar , Coy Pilar , Avilés Manuel , Gadea Joaquin , Romar Raquel
Published: September 2008

Gamete recognition and binding of spermatozoa to the oviduct are carbohydrate-mediated processes in which several glycosidases are thought to have a role, although this has not been demonstrated unequivocally. Oviducal fluid is the biological milieu in which fertilisation and early embryo development take place, but the enzyme composition of oviducal fluid is largely unknown. The aim of the present study was to determine glycosidase activity and protein content in bovine oviducal fluid (bOF) and the volume of fluid collected per oviduct. Oviducts obtained from a slaughterhouse were classified as either in the follicular or luteal phase on the basis of ovarian luteal morphology. Oviducal fluid was aspirated, centrifuged and the volume determined. Samples were then frozen until assay. Substrates conjugated to 4-methylumbelliferyl were used to screen for the activity of seven glycosidases at pH 7.2. The results indicate that bOF has α-l-fucosidase, β-N-acetyl-glucosaminidase, β-d-galactosidase, α-d-mannosidase and β-N-acetyl-galactosaminidase activity during both phases of the cycle, with the specific activity of the latter two enzymes being higher during the follicular phase. There was no N-acetyl-neuraminidase or α-d-galactosidase activity detected in bOF at either phase of the oestrous cycle at pH 7.2, but activity for both glycosidases was detected at pH 4.4. There were no differences in protein concentration or the volume of bOF collected between the two phases of the cycle. These findings indicate that oviducal fluid exhibits glycosidase activity, with specific variations throughout the oestrous cycle, suggesting that these enzymes play a role in carbohydrate-mediated events.
Neuralized is expressed in the α/β lobes of adult
Drosophila
mushroom bodies and facilitates olfactory long-term memory formation
Authors: Pavlopoulos Elias , Anezaki Maria , Skoulakis Efthimios M. C.
Published: September 2008

Memory formation involves multiple molecular mechanisms, the nature and components of which are essential to understand these processes.
Drosophila
is a powerful model to identify genes important for the formation and storage of consolidated memories because the molecular mechanisms and dependence of these processes on particular brain regions appear to be generally conserved. We present evidence that the highly conserved ubiquitin ligase Neuralized (Neur) is expressed in the adult
Drosophila
mushroom body (MB) α/β lobe peripheral neurons and is a limiting factor for the formation of long-term memory (LTM). We show that loss of one copy of
neur
gene results in significant LTM impairment, whereas overexpression of Neur in the peripheral neurons of the α/β lobes of the adult MBs results in a dosage-dependent enhancement of LTM. In contrast, learning, early memories, or anesthesia-resistant memory are not affected. We also demonstrate that the role of Neuralized in LTM formation is restricted within the neurons of the periphery of the α/β lobes, and we suggest that this structural subdivision of the MBs participates in the formation of LTM.
Stem/progenitor cells from bone marrow decrease neuronal death in global ischemia by modulation of inflammatory/immune responses
Authors: Ohtaki Hirokazu , Ylostalo Joni H. , Foraker Jessica E. , Robinson Andrew P. , Reger Roxanne L. , Shioda Seiji , Prockop Darwin J.
Published: September 2008

Human mesenchymal stromal cells (hMSCs) were injected into the hippocampus of adult mice 1 day after transient global ischemia. The hMSCs both improved neurologic function and markedly decreased neuronal cell death of the hippocampus. Microarray assays indicated that ischemia up-regulated 586 mouse genes. The hMSCs persisted for <7 days, but they down-regulated >10% of the ischemia-induced genes, most of which were involved in inflammatory and immune responses. The hMSCs also up-regulated three mouse genes, including the neuroprotective gene
Ym1
that is expressed by activated microglia/macrophages. In addition, the transcriptomes of the hMSC changed with up-regulation of 170 human genes and down-regulation of 54 human genes. Protein assays of the hippocampus demonstrated increased expression in microglia/macrophages of Ym1, the cell survival factor insulin-like growth factor 1, galectin-3, cytokines reflective of a type 2 T cell immune bias, and the major histocompatibility complex II. The observed beneficial effects of hMSCs were largely explained by their modulation of inflammatory and immune responses, apparently by alternative activation of microglia and/or macrophages.
On the best possible remaining term in the Hardy inequality
Authors: Ghoussoub Nassif , Moradifam Amir
Published: September 2008

We give a necessary and sufficient condition on a radially symmetric potential
V
on a bounded domain Ω of ℝ

n

that makes it an admissible candidate for an improved Hardy inequality of the following type. For every ∈
H
1
0
(Ω)

A characterization of the best possible constant
c
(
V
) is also given. This result yields easily the improved Hardy's inequalities of Brezis-Vázquez [Brezis H, Vázquez JL (1997) Blow up solutions of some nonlinear elliptic problems.
Revista Mat Univ Complutense Madrid
10:443–469], Adimurthi
et al.
[Adimurthi, Chaudhuri N, Ramaswamy N (2002) An improved Hardy Sobolev inequality and its applications.
Proc Am Math Soc
130:489–505], and Filippas-Tertikas [Filippas S, Tertikas A (2002) Optimizing improved Hardy inequalities.
J Funct Anal
192:186–233] as well as the corresponding best constants. Our approach clarifies the issue behind the lack of an optimal improvement while yielding the following sharpening of known integrability criteria: If a positive radial function
V
satisfies lim inf
r→o
ln(r)∫
r
o
,
sV
(
s
)
ds
>−∞,then there exists ρ:=ρ(Ω) > 0 such that the above inequality holds for the scaled potential v
ρ
(x)=v(
|x|
ρ
).On the other hand, if lim
r→0
ln(r)∫
r
o
,
sV
(
s
)
ds
=−∞, then there is no ρ > 0 for which the inequality holds for
V
ρ
.
A catechol-
O
-methyltransferase that is essential for auditory function in mice and humans
Authors: Du Xin , Schwander Martin , Moresco Eva Marie Y. , Viviani Pia , Haller Claudia , Hildebrand Michael S. , Pak Kwang , Tarantino Lisa , Roberts Amanda , Richardson Heather , Koob George , Najmabadi Hossein , Ryan Allen F. , Smith Richard J. H. , Müller Ulrich , Beutler Bruce
Published: September 2008

We have identified a previously unannotated catechol-
O
-methyltranferase (COMT), here designated COMT2, through positional cloning of a chemically induced mutation responsible for a neurobehavioral phenotype. Mice homozygous for a missense mutation in
Comt2
show vestibular impairment, profound sensorineuronal deafness, and progressive degeneration of the organ of Corti. Consistent with this phenotype, COMT2 is highly expressed in sensory hair cells of the inner ear. COMT2 enzymatic activity is significantly reduced by the missense mutation, suggesting that a defect in catecholamine catabolism underlies the auditory and vestibular phenotypes. Based on the studies in mice, we have screened DNA from human families and identified a nonsense mutation in the human ortholog of the murine
Comt2
gene that causes nonsyndromic deafness. Defects in catecholamine modification by COMT have been previously implicated in the development of schizophrenia. Our studies identify a previously undescribed COMT gene and indicate an unexpected role for catecholamines in the function of auditory and vestibular sense organs.
A quantitative approach to the development of complex predicates
Authors: Hilpert Martin , Koops Christian
Published: September 2008

This paper traces the historical development of the Swedish Pseudo-Coordination construction with the posture verb sitta “sit”. In Swedish a small number of verbs, including posture verbs such as sitta, are used in coordination with another verb to convey that the described event has an extended duration or is in progress. Quantitative evidence from Swedish historical corpora suggests that the construction has, even after it established itself as a grammatical construction, undergone a number of gradual changes in the course of the past five centuries. As part of the Pseudo-Coordination construction, the verb sitta has changed its argument structure, and the entire construction has increased in syntactic cohesion.
Managing children and adolescents on parenteral nutrition: challenges for the nutritional support team
Authors: Johnson Tracey , Sexton Elaine
Published: September 2008

Managing infants, children and adolescents, ranging from premature infants to 18-year-old adolescents, on parenteral nutrition (PN) is a challenge. The ability of children to withstand starvation is limited and, unlike adults, children require nutrition for growth. PN in children is often required secondary to a congenital bowel problem rather than because of an acquired condition. Conditions requiring PN include motility disorders, congenital disorders of the intestinal epithelium and short-bowel syndrome (SBS). Intestinal failure may be temporary and children with SBS may be weaned from PN. However, other children require permanent PN. There are no comprehensive guidelines for the nutritional requirements of children and adolescents requiring PN. Practice in individual centres is based on clinical experience rather than clinical trials. Requirements are assessed on an individual basis according to age, nutritional status and clinical condition. These requirements need regular review to ensure that they remain appropriate for the changing age and weight of the child. Assessments of intakes use different methods, e.g. reference tables and predictive equations. Complications of PN include infection, accidental damage to, or removal of, the line and cholestatic liver disease. Home parenteral nutrition (HPN) is associated with fewer line infections and allows continuation of nutritional support in a more normal environment, encouraging normal development and participation in family activities. However, having a child at home on HPN is associated with physical and psychological stresses. A feeling of depression, loneliness and social isolation is common amongst children and their families. Home-care services are essential to supporting children at home and should be tailored to, and sensitive to, the individual needs of each family.
Levels of angiogenic factors in pregnancy and post‐partum bleeding
Authors: ESKILD ANNE , JEANSSON STIG , JENUM PÅL A. , STAFF ANNE C. , THADHANI RAVI , KARUMANCHI ANANTH , VATTEN LARS J.
Published: September 2008

We have studied if serum levels of soluble fms‐like tyrosine kinase‐1 (sFlt‐1) and placenta growth factor (PlGF) in pregnancy could predict excess post‐partum bleeding. In 392 normotensive singleton pregnancies, concentrations of sFlt‐1 and PlGF in the first, second, and third trimester were compared between women with and without excess post‐partum bleeding, defined as blood loss volume of at least 500 mL. Mean concentrations of sFlt‐1 were consistently higher in all three trimesters among women who had excess post‐partum bleeding compared to women without this, but significantly higher only in the second trimester. For PlGF, there were no significant differences between the groups. High concentrations of the anti‐angiogenic factor sFlt‐1 in maternal circulation during pregnancy may be associated with an increased risk of excess post‐partum bleeding.
Book Review
Authors: Skaane Per
Published: September 2008

Atlas of Mammography. 3rd edition. By E. Shaw De Paredes. Philadelphia, PA: Lippincott Williams & Wilkins, 2007. ISBN: 978-0-7817-6433-9. 704 pages. Price (hardback): USD 159.00
EPB41L5 functions to post-transcriptionally regulate cadherin and integrin during epithelial–mesenchymal transition
Authors: Hirano Mariko , Hashimoto Shigeru , Yonemura Shigenobu , Sabe Hisataka , Aizawa Shinichi
Published: September 2008

EPB41L5 belongs to the band 4.1 superfamily. We investigate here the involvement of EPB41L5 in epithelial–mesenchymal transition (EMT) during mouse gastrulation. EPB41L5 expression is induced during TGFβ-stimulated EMT, whereas silencing of EPB41L5 by siRNA inhibits this transition. In EPB41L5 mutants, cell–cell adhesion is enhanced, and EMT is greatly impaired during gastrulation. Moreover, cell attachment, spreading, and mobility are greatly reduced by EPB41L5 deficiency. Gene transcription regulation during EMT occurs normally at the mRNA level; EPB41L5 siRNA does not affect either the decrease in E-cadherin or the increase in integrin expression. However, at the protein level, the decrease in E-cadherin and increase in integrin are inhibited in both EPB41L5 siRNA-treated NMuMG cells and mutant mesoderm. We find that EPB41L5 binds p120ctn through its N-terminal FERM domain, inhibiting p120ctn–E-cadherin binding. EPB41L5 overexpression causes E-cadherin relocalization into Rab5-positive vesicles in epithelial cells. At the same time, EPB41L5 binds to paxillin through its C terminus, enhancing integrin/paxillin association, thereby stimulating focal adhesion formation.
Laminins promote postsynaptic maturation by an autocrine mechanism at the neuromuscular junction
Authors: Nishimune Hiroshi , Valdez Gregorio , Jarad George , Moulson Casey L. , Müller Ulrich , Miner Jeffrey H. , Sanes Joshua R.
Published: September 2008

A prominent feature of synaptic maturation at the neuromuscular junction (NMJ) is the topological transformation of the acetylcholine receptor (AChR)-rich postsynaptic membrane from an ovoid plaque into a complex array of branches. We show here that laminins play an autocrine role in promoting this transformation. Laminins containing the α4, α5, and β2 subunits are synthesized by muscle fibers and concentrated in the small portion of the basal lamina that passes through the synaptic cleft at the NMJ. Topological maturation of AChR clusters was delayed in targeted mutant mice lacking laminin α5 and arrested in mutants lacking both α4 and α5. Analysis of chimeric laminins in vivo and of mutant myotubes cultured aneurally demonstrated that the laminins act directly on muscle cells to promote postsynaptic maturation. Immunohistochemical studies in vivo and in vitro along with analysis of targeted mutants provide evidence that laminin-dependent aggregation of dystroglycan in the postsynaptic membrane is a key step in synaptic maturation. Another synaptically concentrated laminin receptor, Bcam, is dispensable. Together with previous studies implicating laminins as organizers of presynaptic differentiation, these results show that laminins coordinate post- with presynaptic maturation.
PSGL-1 engagement by E-selectin signals through Src kinase Fgr and ITAM adapters DAP12 and FcRγ to induce slow leukocyte rolling
Authors: Zarbock Alexander , Abram Clare L. , Hundt Matthias , Altman Amnon , Lowell Clifford A. , Ley Klaus
Published: September 2008

E-selectin binding to P-selectin glycoprotein ligand-1 (PSGL-1) can activate the β2 integrin lymphocyte function-associated antigen-1 by signaling through spleen tyrosine kinase (Syk). This signaling is independent of Gαi-protein–coupled receptors, results in slow rolling, and promotes neutrophil recruitment to sites of inflammation. However, the signaling pathways linking E-selectin engagement of PSGL-1 to Syk activation are unknown. To test the role of Src family kinases and immunoreceptor tyrosine-based activating motif (ITAM)–containing adaptor proteins, we used different gene-deficient mice in flow chamber, intravital microscopy, and peritonitis studies. E-selectin–mediated phosphorylation of Syk and slow rolling was abolished in neutrophils from fgr−/− or hck−/− lyn−/− fgr−/− mice. Neutrophils from Tyrobp−/− Fcrg−/− mice lacking both DAP12 and FcRγ were incapable of sustaining slow neutrophil rolling on E-selectin and intercellular adhesion molecule-1 and were unable to phosphorylate Syk and p38 MAPK. This defect was confirmed in vivo by using mixed chimeric mice. Gαi-independent neutrophil recruitment into the inflamed peritoneal cavity was sharply suppressed in Tyrobp−/− Fcrg−/− mice. Our data demonstrate that an ITAM-dependent pathway involving the Src-family kinase Fgr and the ITAM-containing adaptor proteins DAP12 and FcRγ is involved in the initial signaling events downstream of PSGL-1 that are required to initiate neutrophil slow rolling.
Lophius in the world: a synthesis on the common features and life strategies
Authors: Fariña A. C. , Azevedo M. , Landa J. , Duarte R. , Sampedro P. , Costas G. , Torres M. A. , Cañás L.
Published: September 2008

Fariña, A. C., Azevedo, M., Landa, J., Duarte, R., Sampedro, P., Costas, G., Torres, M. A., and Cañás, L. 2008. Lophius in the world: a synthesis on the common features and life strategies. – ICES Journal of Marine Science, 65: 1272–1280. Seven species of Lophius are known worldwide, six in the Atlantic Ocean and just one in the Northwest Pacific. The genus supports valuable fisheries (except for Lophius vaillanti), most for a long time, though the exploitation of Lophiusgastrophysus along the coast of Brazil is relatively recent. The manuscript reviews the current knowledge of phylogeographic and biological traits of Lophius species, pointing out common aspects in the life histories. Within the Lophiidae, the genus Lophius is phylogenetically the most derived, vicariance and dispersal having played a significant role in driving speciation. Life histories seem to have followed similar adaptive processes from a common ancestor along with similar environmental characteristics. The genetic structure of populations is poorly known, and usually, genetic differentiation is limited. Life-history aspects (age, growth, reproductive cycle, early stages, and feeding ecology) are addressed, and fisheries are reviewed. However, knowledge of many aspects of the biology and ecology (e.g. validation of the growth pattern, maturation processes, spawning areas and periodicity, recruitment processes, mortality, stock identification, and habitat needs) remains limited.
Systematic conservation planning in the Mediterranean: a flexible tool for the identification of no-take marine protected areas
Authors: Maiorano Luigi , Bartolino Valerio , Colloca Francesco , Abella Alvaro , Belluscio Andrea , Carpentieri Paolo , Criscoli Alessandro , Jona Lasinio Giovanna , Mannini Alessandro , Pranovi Fabio , Reale Bruno , Relini Giulio , Viva Claudio , Ardizzone Gian Domenico
Published: September 2008

Maiorano, L., Bartolino, V., Colloca, F., Abella, A., Belluscio, A., Carpentieri, P., Criscoli, A., Jona Lasinio, G., Mannini, A., Pranovi, F., Reale, B., Relini, G., Viva, C., and Ardizzone, G. D. 2009. Systematic conservation planning in the Mediterranean: a flexible tool for the identification of no-take marine protected areas. – ICES Journal of Marine Science, 66: 137–146. We propose the use of systematic conservation planning in the Mediterranean context for the identification of no-take marine protected areas (NTMPAs). We suggest a logical framework that should be used for the identification of areas to be targeted for multispecies, spatially explicit conservation actions. Specifically, we propose seven steps: (i) definition of the study area; (ii) selection of the species or habitats to be considered; (iii) definition of the planning units; (iv) measurement of the fishing effort; (v) definition of the conservation targets; (vi) review of the existing conservation areas; (vii) selection of additional NTMPAs. Moreover, we consider the potential impact of different conservation plans on existing fishing vessels. A working example is presented, focusing on a limited number of species and on a limited study area. This framework can be easily expanded to include datasets of different origin and to accommodate larger spatial scales. Such a process involves major data-collection and capacity-building elements, and conservation of productive commercial fisheries must be a priority.
Monkfish/anglerfish across the world; common problems and common solutions: an introduction to papers presented at the ICES Theme Session in September 2007
Authors: Maguire Jean-Jacques , Pereda Pilar , Duarte Rafael , Dobby Helen , Azevedo Manuela
Published: September 2008

Maguire, J-J., Pereda, P., Duarte R., Dobby, H., and Azevedo, M. 2008. Monkfish/anglerfish across the world; common problems and common solutions: An introduction to papers presented at the ICES Theme Session in September 2007. – ICES Journal of Marine Science, 65: 1270–1271.
What Causes Genetic Disorders?
Authors: Simon Chantal , Farndon Peter
Published: September 2008

Genetics is the science of inheritance. It aims to understand the mechanism by which the blueprints for life are passed through generations and how variations in these blueprints are essential for evolution, yet can cause disease. This article aims to define the major types of genetic disorder and provide examples of each.
The Role of the Practice Manager
Authors: Bateman Cathryn
Published: September 2008

The role of the practice manager has probably been somewhat evolutionary, primarily due to the growing administrative demands placed upon general practitioners.
Human cytomegalovirus interferes with signal transducer and activator of transcription (STAT) 2 protein stability and tyrosine phosphorylation
Authors: Le Vu Thuy Khanh , Trilling Mirko , Wilborn Manuel , Hengel Hartmut , Zimmermann Albert
Published: September 2008

We have investigated the role of signal transducer and activator of transcription (STAT) 2 during human cytomegalovirus (HCMV) replication and found that protein levels of STAT2 are downregulated. STAT2 downregulation was observed in HCMV clinical isolates and laboratory strains with the exception of strain Towne. The HCMV-induced loss of STAT2 protein occurred despite an increased accumulation ofSTAT2mRNA; it required HCMV early gene expression. The decrease in STAT2 was sensitive to proteasome inhibition, suggesting degradation of STAT2 via the ubiquitin proteasome pathway. Notably, pUL27, the HCMV homologue of the mouse CMV pM27 protein, which mediates the selective proteolysis of STAT2, did not induce STAT2 downregulation. Moreover, preceding STAT2 degradation, alpha/beta interferon (IFN)-receptor-mediated tyrosine phosphorylation of STAT2 was inhibited in HCMV-infected cells. This effect was paralleled by impaired tyrosine activation of STAT1 and STAT3. Accordingly, IFNs affected the replication efficiency of STAT2 degrading and non-degrading HCMV strains to a similar degree. In summary, HCMV abrogates IFN receptor signalling at multiple checkpoints by independent mechanisms includingUL27-independent degradation of STAT2 and a preceding blockade of STAT2 phosphorylation.
Four novel human betapapillomaviruses of species 2 preferentially found in actinic keratosis
Authors: Vasiljević Nataša , Hazard Kristina , Dillner Joakim , Forslund Ola
Published: September 2008

Recent studies have suggested an association between human papillomaviruses (HPVs), particularly species 2 members of the genusBetapapillomavirus, and squamous cell carcinoma (SCC) of the skin. As most of these viruses are uncharacterized, molecular characterization and epidemiology are needed to advance our understanding of their significance in carcinogenesis. This study determined the complete genomes of four betapapillomaviruses of species 2 from skin lesions designated HPV-107, -110 and -111 and FA75[KI88-03], an isolate of an unpublished HPV type, and analysed their prevalence and viral loads in biopsies from SCC, actinic keratosis (AK), basal cell carcinoma, seborrhoeic keratosis and the healthy skin of 263 immunocompetent patients by HPV type-specific real-time PCR assays. Seventeen patients (6.5 %) harboured at least one of the four HPV types in their lesion, whereas seven patients (2.7 %) harboured one or more of the HPV types in healthy skin. Overall, the four viruses were more common in AK than in healthy skin (odds ratio 5.0, 95 % confidence interval 1.4–17.5), but the prevalence and viral loads were low. This characterization of HPV-107, -110 and -111 and FA75[KI88-03] expands the heterogeneity of members of species 2 of the genusBetapapillomavirus. However, as these types were found in only a few samples and in low amounts, a possible role in carcinogenesis remains elusive.
Molecular characterization of a novel adult diarrhoea rotavirus strain J19 isolated in China and its significance for the evolution and origin of group B rotaviruses
Authors: Jiang Shengjun , Ji Shaozhong , Tang Qing , Cui Xiaoying , Yang Hongyang , Kan Biao , Gao Shouyi
Published: September 2008

The complete genome of a novel adult diarrhoea rotavirus strain J19 was cloned and sequenced using an improved single-primer sequence-independent method. The complete genome is 17 961 bp and is AU-rich (66.49 %). Northern blot analysis and genomic sequence analysis indicated that segments 1–11 encode 11 viral proteins, respectively. Protein alignments with the corresponding proteins of J19 with B219, and groups A, B and C rotaviruses, produced higher per cent sequence identities to B219. Among groups A, B and C rotaviruses, 10 proteins from group B rotaviruses exhibited slightly higher amino acid sequence identity to the J19 proteins, but proteins of J19 showed low amino acid sequence identity with groups A and C rotaviruses. Construction of unrooted phylogenetic trees using a set of known proteins and representatives of three known rotavirus groups revealed that six structural proteins were positioned close to B219 and the basal nodes of groups A, B and C lineages, although with a preferred association with group B lineages. Phylogenetic analysis of the five non-structural proteins showed a similar trend. The results of the serological analysis, protein sequence analysis and phylogenetic analysis suggested that J19 would be a novel rotavirus strain with great significance to the evolution and origin of group B rotaviruses.
A single amino acid change in a geminiviral Rep protein differentiates between triggering a plant defence response and initiating viral DNA replication
Authors: Jin Mingfei , Li Chunyang , Shi Yan , Ryabov Eugene , Huang Jing , Wu Zirong , Fan Zaifeng , Hong Yiguo
Published: September 2008

We have devised anin plantasystem for functional analysis of the replication-associated protein (Rep) of African cassava mosaic virus (ACMV). Using this assay and PCR-based random mutagenesis, we have identified an ACMV Rep mutant that failed to trigger the hypersensitive response (HR), but had an enhanced ability to initiate DNA replication. The mutant Rep–green fluorescent protein (GFP) fusion protein was localized to the nucleus. Sequence analysis showed that the mutatedRepgene had three nucleotide changes (A6→T, T375→G and G852→A); only the A6→T transversion resulted in an amino acid substitution (Arg to Ser), which is at the second residue in the 358 amino acid ACMV Rep protein. Our results indicate that a single amino acid can alter the differential ability of ACMV Rep to trigger the host-mediated HR defence mechanism and to initiate viral DNA replication. The implications of this finding are discussed in the context of plant–virus interactions.
In Semliki Forest virus encephalitis, antibody rapidly clears infectious virus and is required to eliminate viral material from the brain, but is not required to generate lesions of demyelination
Authors: Fragkoudis Rennos , Ballany Catherine M. , Boyd Amanda , Fazakerley John K.
Published: September 2008

Semliki Forest virus (SFV) infection of the laboratory mouse provides a well-characterized tractable system to study the pathogenesis of virus encephalitis and virus induced demyelination. In μMT mice, which have no antibodies, infectious virus persisted in both the serum and the brain for several weeks, indicating that antibodies are required to eliminate infectious virus. In immunocompetent mice, virus infectivity in the brain was undetectable after the first week of infection, but virus RNA levels declined slowly. Following SFV infection, lesions of demyelination were present in the brains of both immunocompetent and μMT mice, indicating that antibodies are not required to generate lesions of demyelination.
Molecular characterization of medically important viruses of the genus Orthobunyavirus
Authors: Lambert Amy J. , Lanciotti Robert S.
Published: September 2008

We have characterized the full-length S segment RNA sequences of five human pathogens of the virus family Bunyaviridae, genus Orthobunyavirus. S segment sequences of Fort Sherman, Shokwe and Xingu viruses of the Bunyamwera serogroup, as well as those of Bwamba and Pongola viruses of the Bwamba serogroup, are described. S segment sequences of Bwamba and Pongola viruses represent the first nucleotide sequences characterized for viruses of the Bwamba serogroup. The described molecular and phylogenetic analyses of these and other selected viruses of the genus Orthobunyavirus reveal that a close sequence similarity is shared between the African Bwamba and the predominantly North American and European California serogroups of the genus Orthobunyavirus.
Mapping and characterization of visna/maedi virus cytotoxic T-lymphocyte epitopes
Authors: Wu Changxin , Barbezange Cyril , McConnell Ian , Blacklaws Barbara A.
Published: September 2008

CD8+cytotoxic T-lymphocyte (CTL) responses have been shown to be important in the control of human and simian immunodeficiency virus infections. Infection of sheep with visna/maedi virus (VISNA), a related lentivirus, induces specific CD8+CTLin vivo, but the specific viral proteins recognized are not known. To determine which VISNA antigens were recognized by sheep CTL, we used recombinant vaccinia viruses expressing the different genes of VISNA: in six sheep (Finnish Landrace×Dorset crosses, Friesland and Lleyn breeds) all VISNA proteins were recognized except TAT. Two sheep, shown to share major histocompatibility complex (MHC) class I alleles, recognized POL and were used to map the epitope. Thepolgene is 3267 bp long encoding 1088 aa. By using recombinant vaccinia viruses a central portion (nt 1609–2176, aa 537–725) was found to contain the CTL epitope and this was mapped with synthetic peptides to a 25 aa region (aa 612–636). When smaller peptides were used, a cluster of epitopes was detected: at least three epitopes were present, at positions 612–623: DSRYAFEFMIRN; 620–631: MIRNWDEEVIKN; and 625–635: EEVIKNPIQAR. A DNA-prime-modified vaccinia virus Ankara (MVA)-boost strategy was employed to immunize four sheep shown to share MHC class I allele(s) with the sheep above. Specific CTL activity developed in all the immunized sheep within 3 weeks of the final MVA boost although half the sheep showed evidence of specific reactivity after the DNA-prime immunizations. This is the first report, to our knowledge, of induction of CTL by a DNA-prime-boost method in VISNA infection.
Mapping the immune response to the outer domain of a human immunodeficiency virus-1 clade C gp120
Authors: Chen Hongying , Xu Xiaodong , Lin Hsin-Hui , Chen Ssu-Hsien , Forsman Anna , Aasa-Chapman Marlen , Jones Ian M.
Published: September 2008

The outer domain (OD) of human immunodeficiency virus (HIV)-1 gp120 represents an attractive, if difficult, target for a beneficial immune response to HIV infection. Unlike the entire gp120, the OD is structurally stable and contains the surfaces that interact with both the primary and secondary cellular receptors. The primary strain-specific neutralizing target, the V3 loop, lies within the OD, as do epitopes for two cross-reactive neutralizing monoclonal antibodies (mAbs), b12 and 2G12, and the contact sites for a number of inhibitory lectins. The OD is poorly immunogenic, at least in the context of complete gp120, but purposeful OD immunization can lead to a substantial antibody response. Here, we map the antibody generated following immunization with a clade C OD. In contrast to published data for the clade B OD, the majority of the polyclonal response to the complete clade C OD is to the V3 loop; deletion of the loop substantially reduces immunogenicity. When the loop sequence was substituted for the epitope for 2F5, a well-characterized human cross-neutralizing mAb, a polyclonal response to the epitope was generated. A panel of mAbs against the clade C OD identified two mAbs that reacted with the loop and were neutralizing for clade C but not B isolates. Other mAbs recognized both linear and conformational epitopes in the OD. We conclude that, as for complete gp120, V3 immunodominance is a property of OD immunogens, that the responses can be neutralizing and that it could be exploited for the presentation of other epitopes.
Resistance to human immunodeficiency virus type 1 (HIV-1) generated by lentivirus vector-mediated delivery of the CCR5Δ32 gene despite detectable expression of the HIV-1 co-receptors
Authors: Jin Qingwen , Marsh Jon , Cornetta Kenneth , Alkhatib Ghalib
Published: September 2008

It has previously been demonstrated that there are two distinct mechanisms for genetic resistance to human immunodeficiency virus type 1 (HIV-1) conferred by theCCR5Δ32gene: the loss of wild-type CCR5 surface expression and the generation of CCR5Δ32 protein, which interacts with CXCR4. To analyse the protective effects of long-term expression of the CCR5Δ32 protein, recombinant lentiviral vectors were used to deliver theCCR5Δ32gene into human cell lines and primary peripheral blood mononuclear cells that had been immortalized by human T-cell leukemia virus type 1. Blasticidin S-resistant cell lines expressing the lentivirus-encodedCCR5Δ32showed a significant reduction in HIV-1 Env-mediated fusion assays. It was shown that CD4+T lymphocytes expressing the lentivirus-encodedCCR5Δ32gene were highly resistant to infection by a primary but not by a laboratory-adapted X4 strain, suggesting different infectivity requirements. In contrast to previous studies that analysed the CCR5Δ32 protective effects in a transient expression system, this study showed that long-term expression of CCR5Δ32 conferred resistance to HIV-1 despite cell-surface expression of the HIV co-receptors. The results suggest an additional unknown mechanism for generating the CCR5Δ32 resistance phenotype and support the hypothesis that the CCR5Δ32 protein acts as an HIV-suppressive factor by altering the stoichiometry of the molecules involved in HIV-1 entry. The lentiviral–CCR5Δ32 vectors offer a method of generating HIV-resistant cells by delivery of theCCR5Δ32gene that may be useful for stem cell- or T-cell-based gene therapy for HIV-1 infection.
Two out of the 11 genes of an unusual human G6P[6] rotavirus isolate are of bovine origin
Authors: Matthijnssens Jelle , Rahman Mustafizur , Van Ranst Marc
Published: September 2008

In 2003, we described the first human G6P[6] rotavirus strain (B1711). To investigate further the molecular origin of this strain and to determine the possible reassortments leading to this new gene constellation, the complete genome of strain B1711 was sequenced. SimPlot analyses were conducted to compare strain B1711 with other known rotavirus gene segments, and phylogenetic dendrograms were constructed to analyse the origin of the eleven genome segments of strain B1711. Our analysis indicated that strain B1711 acquired its VP1-, VP2-, VP4-, VP6- and NSP1–5-encoding gene segments from human DS-1-like P[6] rotavirus strains, and its VP3 and VP7 gene segments from a bovine rotavirus strain through reassortment. The introduction of animal–human reassortant strains, which might arise in either of the hosts, into the human rotavirus population is an important mechanism for the generation of rotavirus diversity, and might be a challenge for the current rotavirus vaccines and vaccines under development.