bluuebunny/crossref_metadata_2025 huggingface dataset:
Gero Miesenböck: instructing the nervous system
Authors: Wells William A.
Published: May 2007

Gero Miesenböck uses light and genetically encoded sensors and actuators to observe and control neural activity. Having caused headless flies to fly at will, he is set to understand how the nervous system encodes behavior.
αvβ3 and α5β1 integrin recycling pathways dictate downstream Rho kinase signaling to regulate persistent cell migration
Authors: White Dominic P. , Caswell Patrick T. , Norman Jim C.
Published: May 2007

Accumulating evidence suggests that integrin recycling regulates cell migration. However, the lack of reagents to selectively target the trafficking of individual heterodimers, as opposed to endocytic transport as a whole, has made it difficult to define the contribution made by particular recycling pathways to directional cell movement. We show that autophosphorylation of protein kinase D1 (PKD1) at Ser916 is necessary for its association with αvβ3 integrin. Expression of PKD1916A or the use of mutants of β3 that do not bind to PKD1 selectively inhibits short-loop, Rab4-dependent recycling of αvβ3, and this suppresses the persistence of fibroblast migration. However, we report that short-loop recycling does not directly contribute to fibroblast migration by moving αvβ3 to the cell front, but by antagonizing α5β1 recycling, which, in turn, influences the cell's decision to migrate with persistence or to move randomly.
Secreted gliomedin is a perinodal matrix component of peripheral nerves
Authors: Eshed Yael , Feinberg Konstantin , Carey David J. , Peles Elior
Published: May 2007

The interaction between gliomedin and the axonodal cell adhesion molecules (CAMs) neurofascin and NrCAM induces the clustering of Na+ channels at the nodes of Ranvier. We define new interactions of gliomedin that are essential for its clustering activity. We show that gliomedin exists as both transmembrane and secreted forms that are generated by proteolytic cleavage of the protein, and that only the latter is detected at the nodes of Ranvier. The secreted extracellular domain of gliomedin binds to Schwann cells and is incorporated into the extracellular matrix (ECM) in a heparin-dependent manner, suggesting the involvement of heparan sulfate proteoglycans (HSPGs). Furthermore, we show that the N-terminal region of gliomedin serves as an oligomerization domain that mediates self-association of the molecule, which is required for its binding to neurofascin and NrCAM. Our results indicate that the deposition of gliomedin multimers at the nodal gap by binding to HSPGs facilitates the clustering of the axonodal CAMs and Na+ channels.
Identification of long intergenic region sequences involved in maize streak virus replication
Authors: Willment Janet A. , Martin Darrin P. , Palmer Kenneth E. , Schnippenkoetter Wendelin H. , Shepherd Dionne N. , Rybicki Edward P.
Published: May 2007

The maincis-acting control regions for replication of the single-stranded DNA genome of maize streak virus (MSV) are believed to reside within an approximately 310 nt long intergenic region (LIR). However, neither the minimum LIR sequence required nor the sequence determinants of replication specificity have been determined experimentally. There are iterated sequences, or iterons, both within the conserved inverted-repeat sequences with the potential to form a stem–loop structure at the origin of virion-strand replication, and upstream of therepgene TATA box (therep-proximal iteron or RPI). Based on experimental analyses of similar iterons in viruses from other geminivirus genera and their proximity to known Rep-binding sites in the distantly related mastrevirus wheat dwarf virus, it has been hypothesized that the iterons may be Rep-binding and/or -recognition sequences. Here, a series of LIR deletion mutants was used to define the upper bounds of the LIR sequence required for replication. After identifying MSV strains and distinct mastreviruses with incompatible replication-specificity determinants (RSDs), LIR chimaeras were used to map the primary MSV RSD to a 67 nt sequence containing the RPI. Although the results generally support the prevailing hypothesis that MSV iterons are functional analogues of those found in other geminivirus genera, it is demonstrated that neither the inverted-repeat nor RPI sequences are absolute determinants of replication specificity. Moreover, widely divergent mastreviruses cantrans-replicate one another. These results also suggest that sequences in the 67 nt region surrounding the RPI interact in a sequence-specific manner with those of the inverted repeat.
Local delivery of beta interferon using an adeno-associated virus type 5 effectively inhibits adjuvant arthritis in rats
Authors: Adriaansen J. , Fallaux F. J. , de Cortie C. J. , Vervoordeldonk M. J. , Tak P. P.
Published: May 2007

Beta interferon (IFN-β) is a cytokine with potent immunomodulatory properties and has been described as a promising therapeutic molecule for the treatment of rheumatoid arthritis (RA). IFN-β was previously overexpressed intra-articularly using an adenoviral vector in rats with adjuvant arthritis (AA) as a model of RA. This effect was powerful, albeit transient due to the vector chosen. Therefore, in the context of pre-clinical development, a delivery vector optimized for intra-articular gene transfer, recombinant adeno-associated virus type 5 (rAAV5), was selected. To exert an optimal effect, protein production should parallel the course of the disease. For this reason, the gene for IFN-β was placed under the control of an inflammation-responsive [nuclear factor (NF)-κB] promoter. After intra-articular injection of the rAAV5 constructs in rats with AA, local transcription of the transgene and production of the IFN-β protein was found, leading to a pronounced and sustained effect on paw swelling when the expression was under the control of the NF-κB-responsive promoter. Additionally, a significant beneficial effect was observed on proteoglycan depletion and erosions. Thus, intra-articular overexpression of IFN-β using a rAAV5 vector exhibits potential as an innovative therapy for the treatment of RA.
Molecular biology of potexviruses: recent advances
Authors: Verchot-Lubicz Jeanmarie , Ye Chang-Ming , Bamunusinghe Devinka
Published: May 2007

Recent advances in potexvirus research have produced new models describing virus replication, cell-to-cell movement, encapsidation,Rgene-mediated resistance and gene silencing. Interactions between distant RNA elements are a central theme in potexvirus replication. The 5′ non-translated region (NTR) regulates genomic and subgenomic RNA synthesis and encapsidation, as well as virus plasmodesmal transport. The 3′ NTR regulates both plus- and minus-strand RNA synthesis. How the triple gene-block proteins interact for virus movement is still elusive. As the potato virus X (PVX) TGBp1 protein gates plasmodesmata, regulates virus translation and is a suppressor of RNA silencing, further research is needed to determine how these properties contribute to propelling virus through the plasmodesmata. Specifically, TGBp1 suppressor activity is required for virus movement, but how the silencing machinery relates to plasmodesmata is not known. The TGBp2 and TGBp3 proteins are endoplasmic reticulum (ER)-associated proteins required for virus movement. TGBp2 associates with ER-derived vesicles that traffic along the actin network. Future research will determine whether the virus-induced vesicles are cytopathic structures regulating events along the ER or are vehicles carrying virus to the plasmodesmata for transfer into neighbouring cells. Efforts to assemble virionsin vitroidentified a single-tailed particle (STP) comprising RNA, coat protein (CP) and TGBp1. It has been proposed that TGBp1 aids in transport of virions or STP between cells and ensures translation of RNA in the receiving cells. PVX is also a tool for studyingAvr–Rgene interactions and gene silencing in plants. The PVX CP is the elicitor for theRxgene. Recent reports of the PVX CP reveal how CP interacts with theRxgene product.
Camelpox virus encodes a schlafen-like protein that affects orthopoxvirus virulence
Authors: Gubser Caroline , Goodbody Rory , Ecker Andrea , Brady Gareth , O'Neill Luke A. J. , Jacobs Nathalie , Smith Geoffrey L.
Published: May 2007

Camelpox virus (CMLV) gene 176R encodes a protein with sequence similarity to murine schlafen (m-slfn) proteins. In vivo, short and long members of the m-slfn family inhibited T-cell development, whereas in vitro, only short m-slfns caused arrest of fibroblast growth. CMLV 176 protein (v-slfn) is most closely related to short m-slfns; however, when expressed stably in mammalian cells, v-slfn did not inhibit cell growth. v-slfn is a predominantly cytoplasmic 57 kDa protein that is expressed throughout infection. Several other orthopoxviruses encode v-slfn proteins, but the v-slfn gene is fragmented in all sequenced variola virus and vaccinia virus (VACV) strains. Consistent with this, all 16 VACV strains tested do not express a v-slfn detected by polyclonal serum raised against the CMLV protein. In the absence of a small animal model to study CMLV pathogenesis, the contribution of CMLV v-slfn to orthopoxvirus virulence was studied via its expression in an attenuated strain of VACV. Recombinant viruses expressing wild-type v-slfn or v-slfn tagged at its C terminus with a haemagglutinin (HA) epitope were less virulent than control viruses. However, a virus expressing v-slfn tagged with the HA epitope at its N terminus had similar virulence to controls, implying that the N terminus has an important function. A greater recruitment of lymphocytes into infected lung tissue was observed in the presence of wild-type v-slfn but, interestingly, these cells were less activated. Thus, v-slfn is an orthopoxvirus virulence factor that affects the host immune response to infection.
Orf virus interleukin-10 inhibits cytokine synthesis in activated human THP-1 monocytes, but only partially impairs their proliferation
Authors: Wise Lyn , McCaughan Catherine , Tan Chee Keong , Mercer Andrew A. , Fleming Stephen B.
Published: May 2007

The sheep parapoxvirus orf virus (ORFV) induces acute, pustular skin lesions in humans. ORFV encodes an orthologue of interleukin-10 (IL-10) that, whilst it closely resembles ovine IL-10 (91 % amino acid identity), shows only 75 % amino acid identity to human IL-10 (hIL-10). The anti-inflammatory potential of ORFV IL-10 in human ORFV infection was investigated by examining its immunosuppressive effects on THP-1 monocytes. ORFV IL-10 and hIL-10 were shown to have equivalent inhibitory effects on the synthesis of proinflammatory cytokines in lipopolysaccharide-activated monocytes, but differed in their abilities to inhibit monocyte proliferation. Structural modelling of ORFV IL-10 revealed differences from hIL-10 in residues predicted to interact with IL-10 co-receptor 2 (IL-10R2), whereas there were very few differences in the residues predicted to interact with IL-10R1. These findings suggest that the partial ability of ORFV IL-10 to inhibit THP-1 monocyte proliferation may be due to the absence of critical residues that mediate the interaction with human IL-10R2.
Point mutations in the potato leafroll virus major capsid protein alter virion stability and aphid transmission
Authors: Kaplan Igor B. , Lee Lawrence , Ripoll Daniel R. , Palukaitis Peter , Gildow Frederick , Gray Stewart M.
Published: May 2007

The coat protein (CP) of potato leafroll virus (PLRV) is the primary component of the capsid, and is a multifunctional protein known to be involved in vector transmission and virus movement within plant hosts, in addition to particle assembly. Thirteen mutations were generated in various regions of the CP and tested for their ability to affect virus–host and virus–vector interactions. Nine of the mutations prevented the assembly of stable virions. These mutants were unable to infect systemically four different host species. Furthermore, although virus replication and translation of the CP were similar for the mutants and wild-type virus in individual plant cells, the translation of the CP readthrough product was affected in several of the mutants. Four of the mutants were able to assemble stable particles and infect host plants systemically, similarly to the wild-type virus; however, two of the mutants were transmitted less efficiently by aphid vectors. Based on a computer-generated model of the PLRV CP, the mutations that prevented virion assembly were associated with subunit interfaces, while the amino acid alterations in the assembly-competent mutants were associated with surface loops. This and previous work indicates that the CP structural model has value in predicting the structural architecture of the virion.
Efficacy and safety of orbital decompression in treatment of thyroid‐associated ophthalmopathy: long‐term follow‐up of 78 patients
Authors: Jernfors Mia , Välimäki Matti J. , Setälä Kirsi , Malmberg Henrik , Laitinen Kalevi , Pitkäranta Anne
Published: May 2007

Objective To study the long‐term results of decompression surgery in patients with thyroid‐associated ophthalmopathy (TAO).

Design and methods Retrospective analysis and clinical re‐examination of TAO patients with comparison to healthy controls in Helsinki University Central Hospital.

Patients Seventy‐eight patients who had undergone orbital decompression due to TAO by transantral or endonasal technique between the years 1985 and 2000 were invited for re‐examination. For comparison, 79 healthy age‐ and sex‐matched controls underwent a similar examination.

Results In comparison to the patients’ preoperative state, proptosis was reduced by 4·7 (2·6) mm (mean (SD)) in the right and 4·4 (2·9) mm in the left globe (P < 0·0001) but did not reach the level of the controls’ globe (P < 0·0001). In the right and left eye of patients, visual acuity improved in 44–55%, remained stable in 27–36% and worsened in 18–20%, but remained significantly worse than in controls (P =0·02–0·001). Prevalence of diplopia was high at the re‐examination (N = 39, 50%), but 54% of those patients considered it less difficult than before the surgery. Among patients and controls, maxillary sinusitis and facial neuralgias were equally common. Sensory disturbances were significantly more frequent in patients (P < 0·0001). In assessment of overall satisfaction with present eye status, the median of the visual analogue scale (VAS) was lower for the patients than the controls (P < 0·0001). The majority of the patients considered the operation very helpful regardless of the technique used.

Conclusion For patients with TOA, orbital decompression seems to be an effective and safe treatment.
Assessment of thyroid function and urinary and breast milk iodine concentrations in healthy newborns and their mothers in Tehran
Authors: Ordookhani Arash , Pearce Elizabeth N. , Hedayati Mehdi , Mirmiran Parvin , Salimi Sanam , Azizi Fereidoun , Braverman Lewis E.
Published: May 2007

Objective  To measure breast milk iodine (MI) and urinary iodine (UI) concentrations in healthy newborns and their nursing mothers from an iodine‐sufficient region to determine adequacy and to relate these parameters to thyroid function tests in mothers and infants.

Design  Cross‐sectional.

Patients  Forty‐eight healthy neonates of 37 to 42 weeks’ gestation with normal cord blood TSH values and their mothers were recruited in Tehran, Iran.

Measurements  Serum thyroid function tests were performed, and maternal and infant urinary iodine excretion, and maternal MI concentration were measured.

Results  Neonatal age was 12·9 ± 3·9 (mean ± SD; range 7–30) days and maternal age was 25·8 ± 5 years. Median (range) UI in neonates was 271 µg/l (57–800) and in mothers was 107 µg/l (20–710). Median (range) MI was 148 µg/l (45–750). Neonatal and maternal UI did not correlate with serum thyroid function tests. UI < 150, 150–230, and > 230 µg/l was found in 20, 12·5, and 67·5% of neonates and 79·1, 14, and 7% of mothers, respectively. MI was < 150, 150–180, and > 180 µg/l in 52·4, 11·9, and 35·7% of mothers, respectively.

Conclusions  Among euthyroid neonates, UI was adequate despite low median maternal UI and MI concentrations. There were no significant correlations between UI or MI and thyroid function tests in the mothers and infants.
Stream temperature and the potential growth and survival of juvenile Oncorhynchus mykiss in a southern California creek
Authors: BOUGHTON DAVID A. , GIBSON MICHAEL , YEDOR ROBERT , KELLEY ELISE
Published: May 2007

1. We asked whether an increase in food supply in the field would increase the ability of fish populations to withstand climate warming, as predicted by certain bioenergetic models and aquarium experiments.

2. We subsidised the in situ food supply of wild juvenile steelhead (Oncorhynchus mykiss) in a small stream near the species’ southern limit. High‐quality food (10% of fish biomass per day) was added to the drift in eight in‐stream enclosures along a naturally‐occurring thermal gradient.

3. The temperatures during the experiment were well below the upper thermal limit for the species (means of enclosures ranged from 15.1 to 16.5 °C). Food supplements had no discernible effect on survival, but raised mean (± SD) specific growth rate substantially, from 0.038 ± 0.135 in controls to 2.28 ± 0.51 in feeding treatments. Food supplements doubled the variation in growth among fish.

4. The mean and variance of water temperature were correlated across the enclosures, and were therefore transformed into principal component scores T1 (which expressed the stream‐wide correlation pattern) and T2 (which expressed local departures from the pattern). Even though T1 accounted for 96% of the variation in temperature mean and variance, it was not a significant predictor of fish growth. T2 was a significant predictor of growth. The predicted time to double body mass in an enclosure with a large T2 score (cool‐variable) was half that in an enclosure with a low T2 score (warm‐stable).

5. Contrary to expectation, temperature effects were neutral, at least with respect to the main axis of variation among enclosures (cool‐stable versus warm‐variable). Along the orthogonal axis (cool‐variable versus warm‐stable), the effect was opposite from expectations, probably because of temperature variation. Subtle patterns of temperature heterogeneity in streams can be important to potential growth of O. mykiss.
Fault slip in the 1997 Manyi, Tibet earthquake from linear elastic modelling of InSAR displacements
Authors: Funning Gareth J. , Parsons Barry , Wright Tim J.
Published: May 2007

The Mw 7.6 1997 Manyi earthquake occurred in an area of central northern Tibet where sparse vegetation coverage and a lack of human habitation provide excellent conditions for Interferometric Synthetic Aperture Radar (InSAR) studies. We use coseismic pairs of radar images acquired by the ESA ERS-2 satellite to construct interferograms of the surface displacement field due to the earthquake. The location and extent of the coseismic fault rupture are mapped using a combination of optical satellite imagery, high-resolution digital topography, interferometric correlation and azimuth offset measurements; in so doing, we are able to relate prominent geomorphic features in the fault zone to bends in the fault.

Using elastic dislocation models consistent with this mapped fault trace, we then test a range of fault geometries and slip conditions to find the combination which best explains the InSAR displacements. Our favoured model contains a reversal in fault dip, approximately halfway along its length, occurring at the location of a restraining bend. Slip on this model fault is heterogeneous, with two areas of peak slip of 7 m or greater, and components of dip-slip displacement which vary significantly along-strike. The success of this model in fitting the data implies that an observed asymmetry in the coseismic interferograms can be explained in terms of the local fault geometry, rather than by using non-linear elastic rheologies as suggested by earlier authors.
Mutations in the MCFD2 gene are predominant among patients with hereditary combined FV and FVIII deficiency (F5F8D) in India
Authors: JAYANDHARAN G. , SPREAFICO M. , VISWABANDYA A. , CHANDY M. , SRIVASTAVA A. , PEYVANDI F.
Published: May 2007

Summary.  Combined FV and FVIII deficiency (F5F8D) is a rare (1:1.000.000) autosomal recessive disorder caused by a defect in the LMAN1 or MCFD2 genes, encoding for a FV and FVIII cargo receptor complex. We report the phenotype and genotype analyses in nine unrelated Indian patients with low FV and FVIII coagulant activity [FV:C, range: 5.6–22.4% and FVIII:C, range: 8.3–27.1%]. Four homozygous mutations, including two frame shift, one missense and one splice site, were identified in all the nine patients. Three of them, a 72‐bp deletion in LMAN1 (c.813_822 + 62del72, p.K272fs), a 35‐bp deletion in MCFD2 (c.210_244del35) and a missence mutation in MCFD2 (p.D122V), identified in four patients, were novel mutations. A previously reported c.149 + 5G > A transition in MCFD2 was identified in the remaining five patients. Haplotype analysis of MCFD2 gene in patients with p.E71fs and c.149 + 5G > A defects suggested an independent origin of both these mutations. The identification of two common mutations (p.E71fs, c.149 + 5G > A) in MCFD2 gene in seven of nine patients, particularly the c.149 + 5G > A (55,6% of patients), suggests that this gene could be the first to be analysed during the genetic diagnosis of F5F8D in this population. This is the first report describing the molecular analysis of a consistent number of F5F8D patients of South Indian origin, a population with a high frequency of such recessive bleeding disorders.
A sensitive mutation‐specific screening technique for GNAS1 mutations in cases of fibrous dysplasia: the first report of a codon 227 mutation in bone
Authors: Idowu B D , Al‐Adnani M , O'Donnell P , Yu L , Odell E , Diss T , Gale R E , Flanagan A M
Published: May 2007

Aims:  To report on the mutation‐specific restriction enzyme digest (MSRED) method using paraffin‐embedded tissue as a means of detecting GNAS1 mutations in fibrous dysplasia (FD), and to determine if any of the reported GNAS1 mutations in endocrine neoplasms, not previously documented in FD, can be found in FD.

Methods and results:  Sixty‐seven cases of extragnathic FD were analysed as two groups, 1997–2002 and 2003–06, chosen because tissue fixation and decalcification methods were more accurately recorded in the latter. MSRED revealed that between 2003 and 2006, 93% of 28 ‘in house’ extragnathic cases harboured a GNAS1 mutation, compared with 75% of 32 cases before 2003. Fixation times of no more than 48 h and decalcification in ethylenediamine tetraacetic acid gave the best results. Of the 56 mutations detected (five gnathic, 51 extragnathic), 32 (57%) were R201H, 21 (38%) were R201C and three (5%) were Q227L. Two Q227L extragnathic cases had unusual clinical/radiological findings. No mutations were detected in osteofibrous dysplasia.

Conclusion:  Detection of GNAS1 mutations by MSRED is a valuable adjunct to the histopathological diagnosis of FD. This is the first report of a Q227L mutation in FD, although it has been previously documented in pituitary adenoma.
Prognostic significance of histopathological parameters in sentinel nodes of melanoma patients
Authors: Satzger I , Völker B , Al Ghazal M , Meier A , Kapp A , Gutzmer R
Published: May 2007

Aims:  The presence of micrometastases in the sentinel lymph node (SLN) is an important prognostic parameter for melanoma patients. The aim was to determine the prognostic relevance of histopathological characteristics of micrometastases in the SLN, which has not been adequately addressed thus far.

Methods and results:  In 169 melanoma patients with positive SLN, histopathological features of the SLN were correlated with overall survival (OS) and relapse‐free survival (RFS). Tumour burden, expansion of melanoma cells in the periphery (infiltration of capsule) and towards the centre of the SLN [tumour penetrative depth (TPN)] were of prognostic significance for OS and RFS on univariate analysis. Multivariate analysis revealed three independent significant parameters which predict a poor prognosis: presence of infiltration of the SLN capsule, TPN ≥ 2 mm and size of the largest tumour deposit ≥ 30 cells.

Conclusions:  Histopathological analysis of SLN allows the identification of patients with a poor prognosis depending on the location of melanoma cells and tumour burden.
DNA mismatch repair protein expression and microsatellite instability in primary mucosal melanomas of the head and neck
Authors: Marani C , Alvino E , Caporali S , Vigili M G , Mancini G , Rahimi S
Published: May 2007

Aims:  To examine the expression of DNA mismatch repair (MMR) proteins and the presence of microsatellite instability (MSI) in seven primary mucosal melanomas of the head and neck (MMHN).

Methods and results:  Haematoxylin and eosin staining and immunohistochemical analysis for routine diagnostic markers and for MMR proteins were performed. Six cases were examined for MSI. Four cases were monomorphous and three cases were pleomorphic type MMHN. Melanocytic markers were positive in all cases. Immunoreactivity for MMR proteins was weak in normal epithelium. The neoplastic tissue in six cases showed positivity for all MMR proteins with different percentages. One case showed weak positivity for hMSH2 and hMSH6 and no immunoreactivity for hMLH1 or hPMS2. Staining intensity was higher in tumour cells than in matched normal mucosa in three cases for hMSH2 and hMLH1 and in two cases for hPMS2. None of the examined cases showed MSI.

Conclusions:  Expression of hMSH2 and hMLH1 proteins was up‐regulated in three cases, whereas in two cases that of hPMS2 was increased. hMSH6 expression was comparable to that of normal cells in all cases. The percentage of positive neoplastic cells and the intensity of staining seemed to be greater in pleomorphic melanomas. Six cases were MMR‐proficient and microsatellite stable.
Respiratory syncytial virus differentially activates murine myeloid and plasmacytoid dendritic cells
Authors: Boogaard Ivette , Van Oosten Marijke , Van Rijt Leonie S. , Muskens Femke , Kimman Tjeerd G. , Lambrecht Bart N. , Buisman Anne‐Marie
Published: May 2007

Respiratory syncytial virus (RSV) is the primary cause of bronchiolitis in young children. Upon infection both T helper 1 (Th1) and Th2 cytokines are produced. Because RSV‐induced Th2 responses have been associated with severe immunopathology and aggravation of allergic reactions, the regulation of the immune response following RSV infection is crucial. In this study we examined the influence of RSV on the activation and function of murine bone marrow‐derived dendritic cells (DCs). RSV induced the expression of maturation markers on myeloid DCs (mDCs) in vitro. The mDCs stimulated with RSV and ovalbumin (OVA) enhanced proliferation of OVA‐specific T cells, which produced both Th1 and Th2 cytokines. In contrast to mDCs, RSV did not induce the expression of maturation markers on plasmacytoid DCs (pDCs), not did it enhance the proliferation of OVA‐specific T cells that were cocultured with pDCs. However, RSV stimulated the production of interferon‐α (IFN‐α) by pDCs. Our findings indicate a clear difference in the functional activation of DC subsets. RSV‐stimulated mDCs may have immunostimulatory effects on both Th1 and Th2 responses, while RSV‐stimulated pDCs have direct antiviral activity through the release of IFN‐α.
A p38 MAP kinase regulates the expression of the Aedes aegypti defensin gene in mosquito cells
Authors: Chen‐Chih Wu R. , Shaio M.‐F. , Cho W.‐L.
Published: May 2007

An Aedes aegypti p38 (Aap38) mitogen‐activated protein kinase was isolated and characterized in this study. The 1761 bp long full‐length Aap38 cDNA encodes an open reading frame of 358 amino acids, exhibiting characteristics of Thr/Tyr dual kinase specificities. We showed that bacteria activate both the kinase activity of Aap38 and the expression of the Aedes aegypti defensin A (AaDefA) gene, which is inhibited by a p38 kinase inhibitor SB203580 and dsRNA interference of Aap38. A similar result was obtained by a reporter construct containing the AaDefA regulatory region linked to Ds‐Red. The lipopolysaccharide‐activated reporter gene was inhibited by SB203580. In addition, Aap38 translocated to the nucleus after lipopolysaccharide induction. Our findings suggest that the p38 protein kinase pathway is involved in the antibacterial peptide synthesis in mosquitoes.
Affordable assays for genotyping single nucleotide polymorphisms in insects
Authors: Black IV W. C. , Vontas J. G.
Published: May 2007

Insect genome projects and DNA sequence databases are providing unprecedented amounts of information about variation at specific nucleotides in protein‐ and RNA‐coding genes. Single nucleotide polymorphisms (SNPs) are abundant in all insect species so far examined and are proving useful in population genetics, linkage mapping and marker‐assisted selection. A number of studies has already identified SNPs associated with insecticide resistance, especially mutations conferring reduced target site sensitivity. Unfortunately, most modern, high‐throughput, automated SNP detection technologies are expensive or require the use of expensive equipment and are therefore not accessible to laboratories on a limited budget or to our colleagues in developing countries. In this review, we provide a chronological and comprehensive list of all SNP methods. We emphasize and explain those techniques in which genotypes can be identified by eye or that only require agarose gel electrophoresis. We provide examples where these techniques have or are currently being applied to insects.
Investigation of intratumoural and peritumoural lymphatics expressed by podoplanin and LYVE‐1 in the hybridoma‐induced tumours
Authors: Ji R.C. , Eshita Y. , Kato S.
Published: May 2007

Tumour‐associated lymphatics contribute to a key component of metastatic spread, however, the biological interaction of tumour cells with intratumoural and peritumoural lymphatics (ITLs and PTLs) has remained unclear. To address this important issue, we have focused on the morphological and molecular aspects of newly formed lymphatics (lymphangiogenesis) and pre‐existing lymphatics in the intratumoural and peritumoural tissues by using a hybridoma‐induced tumour model. In the present study, ITLs with very high vessel density within the tumour mass showed small and flattened contours that varied from non‐solid‐to‐solid tumours, whereas PTLs were relatively disorganized and tortuous, and packed with a cluster of tumour cells at the tumour periphery. Lymphatic endothelial cells (LECs) both in ITLs and PTLs were expressed with LYVE‐1 and podoplanin in various tumour tissues, in which initial lymphatics were extremely extended and dilated. The tumour cells were frequently detected adhering to or penetrating lymphatic walls, especially near the open junctions. In the metastatic tissues, lymphangiogenic vasculatures occurred within the tumour matrix, and collecting PTLs represented abnormal twisty valve leaflets. The Western blot and RT‐PCR analysis showed local variations of LEC proliferating potentials and lymphatic involvement in metastasis by a distinct profile of the protein and mRNA expression by LYVE‐1, podoplanin, Prox‐1 and vascular endothelial growth factor‐3 (VEGFR‐3). These findings indicated that both ITLs and PTLs, including enlarged pre‐existing and newly formed lymphatics, may play a crucial role in metastasis with an active tumour cell adhesion, invasion, migration and implantation.
Development of an experimental model of brain tissue heterotopia in the lung
Authors: Quemelo Paulo Roberto Veiga , Sbragia Lourenço , Peres Luiz Cesar
Published: May 2007

The presence of heterotopic brain tissue in the lung is a rare abnormality. The cases reported thus far are usually associated with neural tube defects (NTD). As there are no reports of experimental models of NTD that present this abnormality, the objective of the present study was to develop a surgical method of brain tissue heterotopia in the lung. We used 24 pregnant Swiss mice divided into two groups of 12 animals each, denoted 17GD and 18GD according to the gestational day (GD) when caesarean section was performed to collect the fetuses. Surgery was performed on the 15th GD, one fetus was removed by hysterectomy and its brain tissue was cut into small fragments and implanted in the lung of its litter mates. Thirty‐four live fetuses were obtained from the 17GD group. Of these, eight (23.5%) were used as control (C), eight (23.5%) were sham operated (S) and 18 (52.9%) were used for pulmonary brain tissue implantation (PBI). Thirty live fetuses were obtained from the females of the 18GD group. Of these, eight (26.6%) were C, eight (26.6%) S and 14 (46.6%) were used for PBI. Histological examination of the fetal trunks showed implantation of GFAP‐positive brain tissue in 85% of the fetuses of the 17GD group and in 100% of those of the 18GD group, with no significant difference between groups for any of the parameters analysed. The experimental model proved to be efficient and of relatively simple execution, showing complete integration of the brain tissue with pulmonary and pleural tissue and thus representing a model that will permit the study of different aspects of cell implantation and interaction.
Impacts of land use change on South‐east Asian forest butterflies: a review
Authors: KOH LIAN PIN
Published: May 2007

South‐east Asia has the highest relative rate of habitat loss and degradation in the humid tropics. The responses of less ‘charismatic’ groups, including butterflies, to habitat disturbance remain relatively poorly understood. Many South‐east Asian butterflies are endemic to the region and face global extinction if current levels of deforestation were to continue.

Here, I highlight South‐east Asia as a region urgently in need of butterfly conservation research and review empirical studies of the responses of South‐east Asian butterflies to land use change. Additionally, I discuss some methodological pitfalls for such studies. Furthermore, I argue for the importance of identifying the ecological correlates of sensitivity of butterfly species to forest modification and the potential biological mechanisms underlying their responses to land use change.

There has been no consensus among previous studies on the effects of land use change on butterfly communities in South‐east Asia. Of the 20 studies I reviewed, seven reported higher species richness/diversity in undisturbed (or the least disturbed) forest than in disturbed habitats, nine reported the opposite trend, three reported no difference and one reported a strong influence of seasonality on the impacts of logging.

Some of these studies may contain inherent methodological biases resulting from the failure to control for sampling effects, the lack of consideration for the spatial scale of analysis and incomplete sampling of the vertical strata in tropical rainforests.

Synthesis and applications. Empirical studies of the effects of land use change on tropical forest insects are sorely lacking from South‐east Asia. Butterflies are an ideal taxonomic group for such investigations. Future studies should be designed carefully to avoid the methodological pitfalls highlighted here. Determining the ecological correlates of sensitivity of butterflies to forest modification is important for the pre‐emptive identification of species of conservation concern and for generating testable hypotheses on the differential responses of species to forest modification. Experimental studies are needed to determine the mechanisms underlying the responses of species to land use change in order to develop effective strategies for the conservation of butterflies in human disturbed landscapes.
Comparative phylogeography of five sympatric Hypseleotris species (Teleostei: Eleotridae) in south‐eastern Australia reveals a complex pattern of drainage basin exchanges with little congruence across species
Authors: Thacker Christine E. , Unmack Peter J. , Matsui Lauren , Rifenbark Neil
Published: May 2007

Aim  To determine biogeographical patterns in five closely related species in the fish genus Hypseleotris, and to investigate the relative roles of drainage divide crossings and movement during lowered sea levels between drainage basins and biogeographical provinces based on the phylogeographical patterns within the group. The high degree of overlap in the distributions and ecology of these species makes them ideal candidates for comparative phylogeographical study.

Location  Eastern, central and south‐eastern Australia.

Methods  A total of 179 Hypseleotris individuals were sequenced from 45 localities for the complete mitochondrial cytochrome b gene and the first 30 base pairs of the threonine transfer RNA for a total of 1170 bp. Phylogenetic relationships were hypothesized using parsimony and Bayesian analyses.

Results  Phylogenetic analysis resolves the five species into three clades. The first corresponds to the species Hypseleotris klunzingeri (Ogilby, 1898); within it two clades are resolved, one consisting of individuals from the Eastern Province (EP), plus two eastern Murray‐Darling Province (MDP) localities, and the other including the remainder of the MDP localities, along with the Lake Eyre Basin (Central Australian Province, CAP) individuals. The other two clades include a mixed Hypseleotris galii (Ogilby, 1898)/Hypseleotris sp. 3 Murray‐Darling clade, with EP and MDP lineages mostly segregated and differentiations in populations spread along the EP, and a mixed Hypseleotris sp. 4 Lake's and Hypseleotris sp. 5 Midgley's clade, with two groups of MDP localities and two CAP lineages indicated, interspersed with EP lineages as well as those from the Northern Province.

Main conclusions  This study is broadly congruent with a previous analysis of Hypseleotris phylogeny, but the previously observed overall relationship of south‐eastern Australian provinces [EP(MDP+CAP)] was not confirmed and is more complicated than hitherto thought. This highlights the necessity of obtaining a sufficient number of sampling localities to identify potential connectivity between populations in order to demonstrate congruent biogeographical patterns. We identified many instances of drainage divide crossings, which were the major means of movement between provinces. Despite the commonness of movement across drainage divides, very few of these were found to be exactly congruent among the species. Most occurred in different places, or if in the same location, apparently at different times, or in at least one case, in opposite directions. Patterns of movement between adjacent coastal drainages were also found to be largely incongruent; when congruence was found the populations involved had quite different genetic divergences.
Involvement of P‐glycoprotein on ivermectin kinetic behaviour in sheep: itraconazole‐mediated changes on gastrointestinal disposition
Authors: BALLENT M. , LIFSCHITZ A. , VIRKEL G. , SALLOVITZ J. , LANUSSE C.
Published: May 2007

Different pharmacological approaches have been used in an attempt to increase the systemic availability of anthelmintic drugs. The comparative effect of the itraconazole (ITZ)‐mediated modulation of P‐glycoprotein (P‐gp) activity on the in vivo kinetic behaviour of ivermectin (IVM) administered by the intravenous (i.v.) and intraruminal (i.r.) routes to sheep was assessed in the current work. Corriedale sheep received IVM (50 μg/kg) by the i.v. route either alone (group A) or co‐administered with the P‐gp modulator ITZ (100 mg orally three times every 12 h) (group B). Animals in groups C and D were intraruminally treated with IVM (50 μg/kg) alone or co‐administered with ITZ (100 mg orally three times every 12 h) respectively. Jugular blood and gastrointestinal tissue samples (animals treated by the i.r. route) were collected. The samples were analysed by HPLC using fluorescence detection. The plasma disposition of IVM given intravenously was unaffected by the presence of ITZ. However, after the i.r. treatment the co‐administration with ITZ resulted in markedly higher IVM plasma concentration profiles compared to the control group. Likewise, the presence of ITZ enhanced the IVM concentration profiles measured in the gastrointestinal mucosal tissues. An ITZ‐induced reduction on the P‐gp efflux activity at the intestinal lining may have accounted for the greater absorption and enhanced systemic availability observed for IVM in the intraruminally treated animals.
Cellular and humoral immune response to a third generation hepatitis B vaccine
Authors: Schumann A. , Fiedler M. , Dahmen U. , Grosse‐Wilde H. , Roggendorf M. , Lindemann M.
Published: May 2007

Summary.  Liver transplantation is often the ultimate option of therapy for chronically hepatitis B virus (HBV) infected patients. Adoptive transfer of HBV immunity with the liver after vaccination of living liver donors (LLD) could be a new approach to prevent reinfection in the recipients. The time to achieve HBV immunity in LLD is usually short (1–2 months). Therefore, we established a short time immunization protocol (four injections in 2 weeks intervals) using Hepimmune, a recombinant vaccine that contains the L, M and S proteins of HBV. We examined cellular and humoral immune responses after immunization with Hepimmune and compared its immunogenicity to that of a standard HBV vaccine containing only the S protein (HBVAXPRO®). Cellular immunity was measured by interferon (IFN)‐γ ELISpot and proliferation assay. HBV‐specific T cells were detectable in the Hepimmune group after the second and in the standard group after the third vaccination. IFN‐γ production of T cells was significantly higher (P < 0.001) after the third vaccination with Hepimmune. Proliferative responses were also significantly (P < 0.01) higher in the Hepimmune group after the second to fourth vaccination. The humoral immune response could already be detected after the first immunization in nine of 15 Hepimmune vaccinated test persons while it was only observed in one of 15 probands of the later group. Titres differed significantly (P < 0.01) following all four vaccinations. Thus, Hepimmune appears to be a good candidate for short time immunization protocols.
The burden of hepatitis C in England
Authors: Sweeting M. J. , De Angelis D. , Brant L. J. , Harris H. E. , Mann A. G. , Ramsay M. E.
Published: May 2007

Summary.  In England, a large number of individuals are infected with the hepatitis C virus (HCV) and may develop future liver complications, such as decompensated cirrhosis and hepatocellular carcinoma (HCC). Estimates of the magnitude of this future burden are required to plan healthcare resources. We have estimated past incidence of HCV infection in England and predict future burden of end‐stage liver disease in the HCV‐infected population. A model of the natural history of HCV as a series of disease stages was constructed. A back‐calculation approach was performed, using the natural history model and data on annual HCC deaths in England from 1996 to 2004 with mention of HCV and hospital episode statistics for end‐stage liver disease with HCV. The number of HCV‐infected people living with compensated cirrhosis is predicted to rise from 3705 [95% credible interval (CrI): 2820–4975] in 2005 to 7550 (95% CrI: 5120–11 640) in 2015. The number of decompensated cirrhosis and/or HCC cases is also predicted to rise, to 2540 (95% CrI: 2035–3310) by 2015. HCV incidence increased during the 1980s, with an annual incidence of 12 650 (95% CrI: 6150–26 450) by 1989. HCV‐related cirrhosis and deaths from HCC in England are likely to increase dramatically within the next decade. If patients are left undiagnosed and untreated, the future burden of the disease on healthcare resources will be substantial.
Early virologic response after peginterferon alpha‐2a plus ribavirin or peginterferon alpha‐2b plus ribavirin treatment in patients with chronic hepatitis C*
Authors: Di Bisceglie A. M. , Ghalib R. H. , Hamzeh F. M. , Rustgi V. K.
Published: May 2007

Abstract  Patients infected with hepatitis C virus (HCV) genotype 1 and with serum HCV RNA concentrations over 800 000 IU/mL have relatively low rates of virologic response to pegylated interferons. The 2 forms of pegylated interferon have different pharmacokinetic profiles, and pilot studies comparing them have yielded varying results. We compared the virologic response to 12 weeks of treatment with peginterferon alpha‐2a plus ribavirin vs peginterferon alpha‐2b plus ribavirin in 380 patients who were infected with HCV genotype 1 and had high viral loads.

We observed no between‐group differences in viral load reduction over time and no differences in the percentage of patients treated with peginterferon alpha‐2a or peginterferon alpha‐2b plus ribavirin who achieved early virologic response (EVR), defined as ≥2‐log reduction in HCV RNA concentration or undetectable HCV RNA at 12 weeks (66%vs 63%). Serum levels of interferon were more frequently below the level of quantitation in patients treated with peginterferon alpha‐2b plus ribavirin (58–68%) than in those treated with peginterferon alpha‐2a plus ribavirin (1–2%). Patients treated with peginterferon alpha‐2b plus ribavirin had higher rates of discontinuation for safety reasons (6%vs 1%).

In conclusion, a substantial percentage of patients infected with HCV genotype 1 and high viral load can achieve EVR when treated with peginterferon and ribavirin. The 2 pegylated interferons showed comparable anti‐HCV activity during the first 12 weeks of treatment when combined with the same doses of ribavirin (1000–1200 mg/day), but discontinuations for safety reasons were higher in the patients treated with peginterferon alpha‐2b plus ribavirin.
The Boundary Value Problem for the Nonlinear Shallow Water Equations
Authors: Antuono Matteo , Brocchini Maurizio
Published: May 2007

We propose an approximate analytical solution of the boundary value problem (BVP) for the nonlinear shallow waters equations. Our work, based on the Carrier and Greenspan [1] hodograph transformation, focuses on the propagation of nonlinear nonbreaking waves over a uniformly plane beach. Available results are briefly discussed with specific emphasis on the comparison between the Initial Value Problem and the BVP; the latter more completely representing the physical phenomenon of wave propagation on a beach. The solution of the BVP is achieved through a perturbation approach solely using the assumption of small waves incoming at the seaward boundary of the domain. The most significant results, i.e., the shoreline position estimation, the actual wave height and velocity at the seaward boundary, the reflected wave height and velocity at the seaward boundary are given for three specific input waves and compared with available solutions.