bluuebunny/crossref_metadata_2025 huggingface dataset:
Coulomb blockade of small Pd clusters
Authors: Oncel Nuri , Hallback Ann-Sofie , Zandvliet Harold J. W. , Speets Emiel A. , Jan Ravoo Bart , Reinhoudt David N. , Poelsema Bene
Published: July 2005

Single-electron tunneling through Au substrate-alkanethiol-Pd cluster-tip junctions is investigated with scanning tunneling spectroscopy. The measured I(V) curves reveal several characteristic features of the Coulomb blockade, namely, the presence of a Coulomb gap and a Coulomb staircase. By using the orthodox theory of single-electron tunneling, the capacitances and resistances of the double junction system as well as the fractional charge are extracted from the experimental data.
Covariant Hamiltonian for the electromagnetic two-body problem
Authors: De Luca Jayme
Published: July 2005

We give a Hamiltonian formalism for the delay equations of motion of the electromagnetic two-body problem with arbitrary masses and with either repulsive or attractive interaction. This dynamical system based on action-at-a-distance electrodynamics appeared 100 years ago and it was popularized in the 1940s by the Wheeler and Feynman program to quantize it as a means to overcome the divergencies of perturbative QED. Our finite-dimensional implicit Hamiltonian is closed and involves no series expansions. As an application, the Hamiltonian formalism is used to construct a semiclassical canonical quantization based on the numerical trajectories of the attractive problem.
Characterizing energy dissipation in single-walled carbon nanotube polycarbonate composites
Authors: Koratkar Nikhil A. , Suhr Jonghwan , Joshi Amit , Kane Ravi S. , Schadler Linda S. , Ajayan Pulickel M. , Bartolucci Steve
Published: July 2005

In this study, single-walled carbon nanotube and bisphenol-A-polycarbonate composite beams were fabricated by a solution mixing process and dynamic (cyclic) load tests were performed to characterize energy dissipation. We report up to an order of magnitude (>1000%) increase in loss modulus of the polycarbonate system with the addition of 2% weight fraction of oxidized single-walled nanotube fillers. We show that the increase in damping is derived from frictional sliding at the nanotube-polymer interfaces. The nanoscale dimensions of the tubes not only result in large interfacial contact area, thereby generating high damping efficiency, but also enable seamless integration of the filler materials into the composite structure.
Linear viscoelasticity in aluminum thin films
Authors: Hyun Seungmin , Hooghan Tejpal K. , Brown Walter L. , Vinci Richard P.
Published: July 2005

We have found that thin aluminum films at room temperature exhibit changes in elastic modulus of 25% or more over periods of an hour under stress by using a novel capacitance bulge system. Furthermore, we have strong evidence that this change is due to linear viscoelasticity. Such large changes in modulus can affect the stability and performance of microelectromechanical systems structures and need to be understood and controlled. These results are also proof that linear viscoelastic analysis is as appropriate for thin metal films as it is for many polymer systems.
Spatially resolved observation of domain-wall propagation in a submicron ferromagnetic NOT-gate
Authors: Zhu Xiaobin , Allwood Dan A. , Xiong Gang , Cowburn Russell P. , Grütter Peter
Published: July 2005

Domain-wall propagation under an external magnetic field in a submicron ferromagnetic ring integrated with a NOT-junction is investigated by magnetic force microscopy and micromagnetic modeling. Within a certain magnetic field range, one head-to-head or tail-to-tail domain wall propagates in the structure. Magnetic fields above this range cause nucleation of additional domain walls in the ring structure while fields below this range are not able to switch the NOT-junction magnetization. This explicitly demonstrates the magnetization reversal, operation, and failure modes of a magnetic NOT-junction.
Deficit in reproduction in polygynously mated females of the monogamous mound-building mouse Mus spicilegus
Authors: Gouat Patrick , Féron Christophe
Published: July 2005

In monogamous mound-building mice Mus spicilegus, facultative polygyny was observed in April, at the beginning of the reproductive season. In order to evaluate the cost of polygyny, we compared, under laboratory conditions, the reproductive success of trios and pairs composed of sister females mated with unrelated brother males. Females in trios were able to tolerate each other during the reproductive period and to reproduce with the same male under spatial constraints. Nevertheless, polygyny had a strong negative effect on the reproductive success of the females. The average number of young per litter was smaller in trio females than in paired females, whereas the interval between two successive litters was higher. As a result, the number of litters and the number of young per time unit were smaller in each of the trio females compared with paired females. Agonistic behaviour being absent in trios, our results strongly suggest that living in trios led to pheromonal production that affected the female physiological state and reproduction. Males of trios did not obtain a better reproductive success than males in pairs.
p63 regulates commitment to the prostate cell lineage
Authors: Signoretti Sabina , Pires Maira M. , Lindauer Meghan , Horner James W. , Grisanzio Chiara , Dhar Sonya , Majumder Pradip , McKeon Frank , Kantoff Philip W. , Sellers William R. , Loda Massimo
Published: July 2005

Molecular mechanisms underlying prostate and urothelial development remain unclear. This situation presents major limitations in identifying the cell type(s) and molecular events involved in the development of prostate and bladder cancer. It has been shown that mice lacking the basal cell marker p63 present several epithelial defects, including epidermis and prostate buds agenesis and urothelial abnormalities. Here, we use the p63–/– mouse as a tool to define cell lineages in the prostate epithelium and urothelium. By complementing p63–/– blastocysts with p63+/+ β-galactosidase (β-gal)-positive ES cells, we show that secretory cells of the prostate originate from p63-positive basal progenitor cells. Importantly, our urogenital sinus transplantation studies demonstrate that p63 prevents intestinal differentiation of the urogenital sinus endoderm and is therefore required to maintain commitment to the prostate cell lineage. Finally, in contrast with the prostate findings, analysis of the urothelium from rescued p63–/– chimeras shows that umbrella (superficial) cells can develop and be maintained independently from p63-positive basal and intermediate cells.
Caveolin-1 expression by means of p38β mitogen-activated protein kinase mediates the antiproliferative effect of carbon monoxide
Authors: Kim Hong Pyo , Wang Xue , Nakao Atsunori , Kim Sung Il , Murase Noriko , Choi Mary E. , Ryter Stefan W. , Choi Augustine M. K.
Published: July 2005

During vascular injury, the proliferation and migration of smooth muscle cells leads to characteristic neointima formation, which can be exacerbated by genetic depletion of caveolin-1 or heme oxygenase 1 (HO-1), and inhibited by carbon monoxide (CO), a by-product of heme oxygenase 1 activity. CO inhibited smooth muscle cell proliferation by activating p38 mitogen-activated protein kinase (MAPK) and p21Waf1/Cip1. Exposure to CO increased caveolin-1 expression in neointimal lesions of injured aorta andin vitroby activating guanylyl cyclase and p38 MAPK. p38β–/–fibroblasts did not induce caveolin-1 in response to CO, and exhibited a diminished basal caveolin-1 expression, which was restored by p38β gene transfer. p38β MAPK down-regulated extracellular signal-regulated protein kinase 1/2 (ERK-1/2), which can repress caveolin-1 transcription. Genetic depletion of caveolin-1 abolished the antiproliferative effect of CO. Thus, we demonstrate that CO, by activating p38β MAPK, up-regulates caveolin-1, which acts as a tumor suppressor protein that mediates the growth inhibitory properties of this gas.
Electrogene transfer of an Epstein–Barr virus‐based plasmid replicon vector containing the diphtheria toxin A gene suppresses mammary carcinoma growth in SCID mice
Authors: Shibata Masa‐Aki , Miwa Yoshihiro , Miyashita Minoru , Morimoto Junji , Abe Hideaki , Otsuki Yoshinori
Published: July 2005

Experimental mammary cancer therapy in mice was conducted using electrogene transfer of a non‐viral EBV‐based plasmid vector (reduced size of the oriP gene), containing the DT‐A gene. Because the EBV‐based plasmid vector exhibits high transfer efficiency and strong persistent transgene expression due to autonomous replication in human cells, it is particularly suitable as a tool for cancer gene therapy. In vitro, 79% of MDA‐MB231 human mammary carcinoma cells died as a result of the EBV‐based vector containing DT‐A (pEB‐DTA) by 48 h after transfection. DNA synthesis was also significantly decreased as compared to levels with a control vector. In vivo, mammary tumors induced by inoculation of SCID mice with MDA‐MB231 cells were subsequently treated by direct injection of pEB‐DTA vector or pEB‐GFP vector as a control once a week for 5 weeks. After each injection, the tumors were subjected to in vivo electrogene transfer. Significantly reduced tumor volumes were observed for the pEB‐DTA group in experimental week 1 and thereafter throughout the study. At necropsy, strong and extent expression of GFP was still observed in tumors receiving pEB‐GFP 6 days after the last electrogene transfer. The ratio of histological necrotic area to viable area was significantly increased in the pEB‐DTA‐treated tumors, where levels of apoptosis were significantly higher than those observed in the pEB‐GFP group. DNA synthesis showed a tendency to decrease in the pEB‐DTA group but this was not significant. The incidence and multiplicity of lung metastasis were similar between the groups. There was also no difference in the density of microvessels between groups. We therefore conclude that the EBV‐based plasmid vector system combined with in vivo electrogene transfer can result in efficient gene transfection and that the non‐viral replicon vector containing DT‐A suppresses tumor growth due to apoptotic cell death in this model. (Cancer Sci 2005; 96: 434–440)
Relationship between diabetes mellitus and adenocarcinoma of the oesophagus and gastric cardia
Authors: RUBENSTEIN J. H. , DAVIS J. , MARRERO J. A. , INADOMI J. M.
Published: July 2005

Background:  Obesity is a risk factor for adenocarcinomas of the oesophagus and gastric cardia. Diabetes mellitus might mediate that association.

Aim:  To estimate the risk of diabetes mellitus on the development of adenocarcinoma of distal oesophagus and gastric cardia beyond that of gastro‐oesophageal reflux disease.

Methods:  A case–control study was performed using a national administrative database of the Veterans Administration.

Results:  A total of 311 cases of cancer and 10 154 controls were identified. Gender, age, and race were risks for cancer. Diabetes was diagnosed in 36% of cases, and 32% of controls (P = 0.15). Diabetic complications were diagnosed in 14% of cases and 13% of controls (P = 0.60). Multiple logistic regression confirmed the absence of an association between cancer and diabetes (odds ratio 1.1, 95% confidence interval 0.8–1.5) or diabetic complications (odds ratio 0.8, 95% confidence interval 0.6–1.3), adjusting for age, gender, and race.

Conclusions:  Within the limitations of this case–control study, there is no evidence of an association between diabetes and adenocarcinoma of the oesophagus or gastric cardia among US veterans with gastro‐oesophageal reflux disease.
α‐fetoprotein response predicts survival benefits of thalidomide in advanced hepatocellular carcinoma
Authors: CHEN L.‐T. , LIU T.‐W. , CHAO Y. , SHIAH H.‐S. , CHANG J.‐Y. , JUANG S.‐H. , CHEN S.‐C. , CHUANG T.‐R. , CHIN Y.‐H. , WHANG‐PENG J.
Published: July 2005

Background: Radiographic measurements do not always reflect the biological response of hepatocellular carcinoma to drug therapy.

Aims: To evaluate the clinical implications of tumour marker (α‐fetoprotein) response in advanced hepatocellular carcinoma patients with thalidomide treatment.

Patients and methods: Forty‐two advanced hepatocellular carcinoma patients with baselineα‐fetoprotein levels above 200 ng/mL and thalidomide therapy were included. Serumα‐fetoprotein levels were measured every 4 weeks.α‐fetoprotein response was defined as a 50% or greater reduction ofα‐fetoprotein levels for 4 or more weeks during treatment. Radiographic response was assessed by World Health Organization criteria; survivals were estimated by Kaplan–Meier method and prognostic factors were assessed by Cox's proportional hazard model.

Results: With intention‐to‐treat analysis, radiographic response andα‐fetoprotein response were obtained in 7% (three of 42, 95% confidence interval: 0–15) and 24% (10 of 42, 95% CI: 10–38) of patients, respectively. All radiographic response was observed inα‐fetoprotein responders. Multivariate analyses showedα‐fetoprotein response was independent prognostic factor for both progression‐free survival (relative risk = 0.394, 95% CI: 0.189–0.820,P = 0.013) and overall survival (relative risk = 0.241, 95% CI: 0.096–0.606,P =0.003), whereas radiographic response was not.

Conclusion: α‐fetoprotein response can more accurately reflect the biological response of advanced hepatocellular carcinoma to thalidomide therapy than radiographic response.
Daily treatment with esomeprazole is superior to that taken on‐demand for maintenance of healed erosive oesophagitis
Authors: SJÖSTEDT S. , BEFRITS R. , SYLVAN A. , HARTHON C. , JÖRGENSEN L. , CARLING L. , MODIN S. , STUBBERÖD A. , TOTH E. , LIND T.
Published: July 2005

Background:  On‐demand therapy with esomeprazole is effective for long‐term treatment of non‐erosive gastro‐oesophageal reflux disease, but it has not been evaluated in erosive gastro‐oesophageal reflux disease.

Aims:  To compare endoscopic and symptomatic remission over a 6‐month period when patients with healed erosive gastro‐oesophageal reflux disease are treated with esomeprazole 20 mg, either once daily or on‐demand.

Methods:  Patients with verified erosive reflux oesophagitis of Los Angeles grades A–D were enrolled. Following 4–8 weeks treatment with esomeprazole 40 mg daily, those who were endoscopically healed and had symptom control during the last week were randomized to maintenance therapy for 6 months with esomeprazole 20 mg, taken either once daily or on‐demand.

Results:  Of 539 enrolled patients, 494 (91%) were healed at 8 weeks and 477 were randomized to maintenance therapy with esomeprazole 20 mg, 243 once daily and 234 on‐demand. After once daily treatment, 81% of patients were still in remission at 6 months, compared with only 58% who took on‐demand treatment (P < 0.0001). A difference in remission was found irrespective of baseline grade of oesophagitis, but it was more pronounced for the more severe grades. There was no difference in overall symptomatic remission between the two treatments, although heartburn was significantly more prevalent in the on‐demand group.

Conclusions:  Once daily esomeprazole 20 mg was better than that taken on‐demand for maintaining healed erosive oesophagitis, regardless of baseline Los Angeles grade.
Continuing challenges in childhood non‐Hodgkin's lymphoma
Authors: Pinkerton Ross
Published: July 2005

Non‐Hodgkin's lymphoma in children includes a number of different pathological subtypes and, with improved diagnostic techniques and better understanding of the natural history of each type, treatment strategies have become more tumour‐specific. Original staging systems are now less useful in determining treatment stratification and there is a need for careful analysis of large cohorts of patients, treated in the same manner, to determine the clinical and biological features that, with current therapies, are of prognostic value. Novel imaging techniques, such as positron emission tomography, and molecular techniques to detect low‐level marrow involvement are likely to be incorporated into new risk grouping. These will be used to determine the extent of initial disease and evaluate more accurately the speed and quality of response to chemotherapy. With high cure rates, it becomes particularly important to minimize late effects of therapy and the introduction of monoclonal antibodies in combination with chemotherapy may provide a method for improving outcome in poor risk groups and reducing sequelae by allowing reduction in chemotherapy in good risk patients.
Efficacy of the interleukin‐2 receptor antagonist basiliximab in steroid‐refractory acute graft‐versus‐host disease
Authors: Schmidt‐Hieber Martin , Fietz Thomas , Knauf Wolfgang , Uharek Lutz , Hopfenmüller Werner , Thiel Eckhard , Blau Igor Wolfgang
Published: July 2005

Acute graft‐versus‐host disease (aGVHD) occurs in up to 80% of patients who undergo allogeneic stem cell transplantation (SCT) and contributes significantly to transplant‐related mortality (TRM). We conducted a prospective phase II trial to assess the efficacy and feasibility of treating steroid‐refractory aGVHD with basiliximab, a chimaeric monoclonal antibody directed against the alpha chain of the interleukin‐2 (IL‐2) receptor. Basiliximab was administered intravenously at a dose of 20 mg on days 1 and 4. Twenty‐three patients were enrolled between October 1999 and July 2004. We found a primary overall response rate of 82·5% with four patients (17·5%) showing a complete response and 15 patients (65%) a partial response. Six patients were again treated successfully with an IL‐2 receptor antagonist because of recurrence of aGVHD. The rates of infections, chronic GVHD, malignancy recurrence and 1‐year TRM following immunosuppression with basiliximab were comparable with those found with other treatment modalities for aGVHD. We conclude that basiliximab is efficient and feasible for steroid‐refractory aGVHD and merits further evaluation.
Clinical and molecular variability in congenital dyserythropoietic anaemia type I
Authors: Tamary Hannah , Dgany Orly , Proust Alexis , Krasnov Tatyana , Avidan Nili , Eidelitz‐Markus Tal , Tchernia Gil , Geneviève David , Cormier‐Daire Valérie , Bader‐Meunier Brigitte , Ferrero‐Vacher Corinne , Munzer Martine , Gruppo Ralph , Fibach Eithan , Konen Osnat , Yaniv Isaac , Delaunay Jean
Published: July 2005

Congenital dyserythropoietic anaemia (CDA) type I is a rare, inherited disorder characterised by ineffective erythropoiesis and macrocytic anaemia. Complex bone disease has only occasionally been associated with this disease. CDA I is caused by mutations in the CDAN1 gene encoding for codanin‐1. Our aim was to characterise the CDAN1 mutation in eight unrelated patients with sporadic CDA I, three of whom had complex bone disease. Six novel mutations in the CDAN1 gene were identified. In two patients, one mutation and in another, both mutations were elusive. No patient was homozygous for a null‐type mutation. However, one patient with complex bone disease was homozygous for a splice‐site mutation (IVS‐12+5G > A). Western blotting revealed that codanin‐1 synthesis was 65% less than the control. Five single nucleotide polymorphisms (SNPs) previously unreported in the literature or the SNP database were also identified. Although the absence of codanin‐1 is probably lethal, the presence of 35% of the protein was compatible with life but was associated with severe clinical manifestations. However, in most patients studied, no correlation could be established between the expected levels of codanin‐1 or the nature of the mutation and the severity of the clinical manifestations.
Zoledronate synergises with imatinib mesylate to inhibit Ph+ primary leukaemic cell growth
Authors: Segawa Hidekazu , Kimura Shinya , Kuroda Junya , Sato Kiyoshi , Yokota Asumi , Kawata Eri , Kamitsuji Yuri , Ashihara Eishi , Yuasa Takeshi , Fujiyama Yoshihide , Ottmann Oliver G. , Maekawa Taira
Published: July 2005

We examined the in vivo effects and safety of the third generation bisphosphonate, zoledronate (ZOL) alone and combined with imatinib mesylate against primary Philadelphia chromosome positive (Ph+) leukaemic cells. ZOL inhibited the prenylation of Rap1A in leukaemic cells in vitro and synergised with imatinib to enhance the survival of mice engrafted with cells from imatinib‐responders, but not from non‐responders because of mutated BCR‐ABL. These findings suggest that the combination of ZOL and imatinib accelerate the eradication of Ph+ clone, resulting in better prognosis of Ph+ leukaemia patients who have not yet acquired mutations.
Improved outcome for children with disseminated adenoviral infection following allogeneic stem cell transplantation
Authors: Kampmann B. , Cubitt D. , Walls T. , Naik P. , Depala M. , Samarasinghe S. , Robson D. , Hassan A. , Rao K. , Gaspar H. , Davies G. , Jones A. , Cale C. , Gilmour K. , Real M. , Foo M. , Bennett‐Rees N. , Hewitt A. , Amrolia P. , Veys Paul
Published: July 2005

Adenovirus (AdV) infections are a frequent cause of morbidity and mortality following allogeneic stem cell transplantation (SCT), and disseminated infection is associated with high mortality, particularly in paediatric SCT. Here, we describe an approach to reduce mortality from adenoviraemia by combining prospective monitoring for the occurrence of adenoviraemia using a sensitive polymerase chain reaction method, early antiviral therapy and prompt withdrawal of immunosuppression. A total of 155 consecutive paediatric SCT procedures were prospectively monitored, of which 113 (73%) transplants involved donors other than matched siblings and 126 (83%) employed T‐cell depletion. Adenoviraemia was detected in 26/155 (17%) transplants and developed exclusively in patients who had received T‐cell‐depleted grafts. Withdrawal of immunosuppression coupled with early antiviral therapy led to resolution of adenoviraemia in 19/26 (81%) patients with only five patients succumbing to disseminate AdV infection. Survival from adenoviraemia was associated with lymphocyte recovery to above 0·3 × 109/l. Mortality was closely linked with the absence of lymphocyte recovery because of profound T‐cell depletion of the graft with CD34+ magnetic‐activated cell sorting. Mortality from disseminated AdV infection was 5/26 (19%) in this study, which is significantly lower than previously reported.
Activity of selective fully human agonistic antibodies to the TRAIL death receptors TRAIL‐R1 and TRAIL‐R2 in primary and cultured lymphoma cells: induction of apoptosis and enhancement of doxorubicin‐ and bortezomib‐induced cell death
Authors: Georgakis Georgios V. , Li Yang , Humphreys Robin , Andreeff Michael , O'Brien Susan , Younes Mamoun , Carbone Antonino , Albert Vivian , Younes Anas
Published: July 2005

Tumour necrosis factor‐related apoptosis‐inducing ligand (TRAIL/Apo2L) is a death protein that preferentially kills tumour cells while sparing normal cells. TRAIL has four exclusive receptors, two of which (TRAIL‐R1, TRAIL‐R2) are death receptors. Both TRAIL/Apo2L and agonistic antibodies to the TRAIL death receptors are currently being explored for cancer therapy. Although the activity of TRAIL/Apo2L in a variety of haematological malignancies has been examined, the activity of anti‐TRAIL receptor agonistic antibodies in primary and cultured lymphoma cells has not. Using two fully human selective agonistic monoclonal antibodies to the TRAIL death receptors TRAIL‐R1 (HGS‐ETR1) and TRAIL‐R2 (HGS‐ETR2) this study demonstrated that both monoclonal antibodies activated caspase‐8 and induced cell death in five of nine human lymphoma cell lines, and induced >10% cell death in 67% and 70%, respectively, of 27 primary lymphoma cells, and >20% cell death in at least one‐thirds of the samples. HGS‐ETR1 and HGS‐ETR2 demonstrated comparable activity in the fresh tumour samples, which was independent of TRAIL receptor surface expression, Bax, cFLIP, or procaspase‐8 expression, or exposure to prior therapy. Furthermore, both antibodies enhanced the killing effect of doxorubicin and bortezomib. Our data demonstrate that HGS‐ETR1 and HGS‐ETR2 monoclonal antibodies can induce cell death in a variety of cultured and primary lymphoma cells, and may have therapeutic value in lymphoma.
188Re or 90Y‐labelled anti‐CD66 antibody as part of a dose‐reduced conditioning regimen for patients with acute leukaemia or myelodysplastic syndrome over the age of 55: results of a phase I–II study
Authors: Ringhoffer Mark , Blumstein Norbert , Neumaier Bernd , Glatting Gerhard , von Harsdorf Stephanie , Buchmann Inga , Wiesneth Markus , Kotzerke Jörg , Zenz Thorsten , Buck Andreas K. , Schauwecker Peter , Stilgenbauer Stephan , Döhner Hartmut , Reske Sven N. , Bunjes Donald
Published: July 2005

In a phase I–II study for patients aged 55–65 years, we employed radioimmunotherapy using an anti‐CD‐66 antibody as part of a dose‐reduced conditioning regimen, which was followed by a T‐cell‐depleted graft. 20 patients with a median age of 63 years suffering from acute leukaemia (n = 17) or myelodysplastic syndrome (n = 3) received the antibody labelled either with 188Rhenium (n = 8) or with 90Yttrium (n = 12) during conditioning. Radioimmunotherapy provided a mean dose of 21·9 (±8·4) Gy to the bone marrow with a significantly higher dose when 90Yttrium was used. Additional conditioning was fludarabine‐based plus anti‐thymocyte globulin in matched related donor transplants (n = 11), or plus melphalan in matched unrelated donor transplants (n = 9). Regimen‐related toxicity was low, with two patients developing three episodes of grade III organ toxicity. All patients engrafted, grade II–IV acute graft‐versus‐host disease (GvHD) was observed in one patient (5%) and chronic GvHD in three patients (15%). The cumulative incidence of non‐relapse mortality was 25%, the cumulative incidence of relapse 55%. The probability of survival was estimated to be 70% at 1 year and 52% at 2 years post‐transplant, although no plateau was reached afterwards. In conclusion, radioimmunotherapy using the anti‐CD66 antibody was feasible and safe in our elderly patient group and provided a high marrow dose.
Infectious pancreatic necrosis in Atlantic salmon, Salmo salar L.
Authors: Roberts R J , Pearson M D
Published: July 2005

A clinical and histopathological review was carried out of 21 outbreaks of acute infectious pancreatic necrosis (IPN) in Scottish Atlantic salmon, Salmo salar L., farms (13 marine and eight fresh water) during 1991–2004. A distinctive syndrome was evident in both post‐smolts in sea water and fry in fresh water, where liver lesions, which had not previously been associated with IPN, became a consistent finding in addition to the more typical pancreatic and intestinal changes. Initial cases were described in post‐smolts in Shetland, but by the end of the period of investigation this type of pathology had extended down the West coast of Scotland and into Ireland. Limited viral strain analysis suggested that similar strains were involved in both fresh water and sea water and that these differed from earlier isolates from rainbow trout, Oncorhynchus mykiss (Walbaum). In fresh water, recovered fish frequently developed a greatly distended intestine associated with accumulation of undigested food. In sea water, after the initial, often significant (50% or more), losses, there were many fish which failed to grow and became chronically emaciated and prone to sea louse infection. Although use of transfer diets containing immune enhancers and the selection of IPN resistant broodstock has reduced losses the disease remains a serious cause of economic loss.
Virulence factors and pathogenicity of Hafnia alvei for gilthead seabream, Sparus aurata L.
Authors: Padilla D , Real F , Gómez V , Sierra E , Acosta B , Déniz S , Acosta F
Published: July 2005

Virulence factors (eae gene, haemolytic capacity, fimbriae, resistance to the bactericidal effect of serum, siderophore production) and pathogenicity for gilthead seabream, Sparus aurata L., were analysed for 23 Hafnia alvei strains. None of the strains used in LD50 studies were lethal for seabream at doses as high as >108 cfu mL−1. In chronic challenge studies differences in severity of the inflammatory response were observed between strains. On the basis of correlation of the inflammatory response to different strains of H. alvei in seabream with those virulence factors studied, it was only possible to establish a positive correlation between pathogenicity and resistance to the bactericidal effect of fish serum. Gilthead seabream is thus a species with considerable resistance to experimental infection with H. alvei. The bacterium does, however, have the capacity to remain viable in seabream for up to 3 months, without any clinical signs. Hafnia alvei is a well‐recognized human and animal pathogen. Thus, as the pathogen can coexist with aquaculture operations, cultured gilthead seabream could represent a risk to human health as a carrier in some circumstances.
Identification and structural characterization of an unusual mycobacterial monomeromycolyl‐diacylglycerol
Authors: Kremer Laurent , De Chastellier Chantal , Dobson Gary , Gibson Kevin J. C. , Bifani Pablo , Balor Stéphanie , Gorvel Jean‐Pierre , Locht Camille , Minnikin David E. , Besra Gurdyal S.
Published: July 2005

Systematic thin layer chromatographic (TLC) analysis of apolar lipids in Mycobacterium kansasii revealed the presence of a previously uncharacterized novel component. The product was ubiquitously found in a panel of M. kansasii clinical isolates, as well as other pathogenic and non‐pathogenic mycobacterial species. TLC analysis of [14C]‐acetate‐ or [14C]‐glycerol‐labelled M. kansasii cultures tentatively assigned the novel product as an unusual triacylglycerol‐related lipid. Subsequent purification, followed by structural determination using 1H‐nuclear magnetic resonance (NMR) and electrospray mass spectrometry (ES/MS), led to the identification of this product as a monomeromycolyl‐diacylglycerol (MMDAG). Treatment of M. kansasii with either isoniazid (INH), a well‐known type II fatty acid synthase (FAS‐II) and mycolic acid biosynthesis inhibitor, or tetrahydrolipstatin (THL), a drug approved for treating obesity, correlated with a reduced incorporation of [14C]‐acetate into both mycolic acids and MMDAG. Addition of INH or THL to the cultures induced major morphological changes and, surprisingly, resulted in an increased number of lipid storage bodies, as determined by electron microscopy. The potent antimycobacterial activity of THL was confirmed against a variety of mycobacterial species, including INH‐susceptible and ‐resistant Mycobacterium tuberculosis strains. Therefore, THL and other β‐lactones may be promising drugs for the development of new antitubercular therapy.
A symbiosis expressed non‐ribosomal peptide synthetase from a mutualistic fungal endophyte of perennial ryegrass confers protection to the symbiotum from insect herbivory
Authors: Tanaka Aiko , Tapper Brian A. , Popay Alison , Parker Emily J. , Scott Barry
Published: July 2005

While much is known about the biosynthesis of secondary metabolites by filamentous fungi their biological role is often less clear. The assumption is these pathways have adaptive value to the organism but often the evidence to support this role is lacking. We provide the first genetic evidence that the fungal produced secondary metabolite, peramine, protects a host plant from insect herbivory. Peramine is a potent insect feeding deterrent synthesized by Epichloë/Neotyphodium mutualistic endophytes in association with their grass hosts. The structure of peramine, a pyrrolopyrazine, suggests that it is the product of a reaction catalysed by a two‐module non‐ribosomal peptide synthetase (NRPS). Candidate sequences for a peramine synthetase were amplified by reverse transcription polymerase chain reaction. Four unique NRPS products were identified, two of which were preferentially expressed in planta. One of these hybridized to known peramine producing strains. This clone was used to isolate an Epichloë festucae cosmid that contained a two‐module NRPS, designated perA. Nine additional genes, which show striking conservation of microsynteny with Fusarium graminearum and other fungal genomes, were identified on the perA‐containing cosmid. Associations between perennial ryegrass and an E. festucae mutant deleted for perA lack detectable levels of peramine. A wild‐type copy of perA complemented the deletion mutant, confirming that perA is a NRPS required for peramine biosynthesis. In a choice bioassay, plant material containing the perA mutant was as susceptible to Argentine stem weevil (ASW) (Listronotus bonariensis) feeding damage as endophyte‐free plants confirming that peramine is the E. festucae metabolite responsible for ASW feeding deterrent activity.
The crystal structures of Lactococcus lactis MG1363 Dps proteins reveal the presence of an N‐terminal helix that is required for DNA binding
Authors: Stillman Timothy J. , Upadhyay Manisha , Norte Valia A. , Sedelnikova Svetlana E. , Carradus Maria , Tzokov Svetomir , Bullough Per A. , Shearman Claire A. , Gasson Michael J. , Williams Colin H. , Artymiuk Peter J. , Green Jeffrey
Published: July 2005

Dps proteins play a major role in the protection of bacterial DNA from damage by reactive oxygen species. Previous studies have implicated the extended lysine‐containing N‐terminal regions of Dps subunits in DNA binding, but this part of the structure has not previously been observed crystallographically. Here the structures of two Dps proteins (DpsA and DpsB) from Lactococcus lactis MG1363 reveal for the first time the presence of an N‐terminal α helix that extends from the core of the Dps subunit. Consequently, the N‐terminal helices are displayed in parallel pairs on the exterior of the dodecameric Dps assemblies. Both DpsA and DpsB bind DNA. Deletion of the DpsA N‐terminal helix impaired DNA binding. The N‐terminal Lys residues of Escherichia coli Dps have been implicated in DNA binding. Replacement of the lactococcal DpsA Lys residues 9, 15 and 16 by Glu did not inhibit DNA binding. However, DNA binding was inhibited by EDTA, suggesting a role for cations in DNA binding. In contrast to E. coli, Bacillus brevis and Mycobacterium smegmatis Dps:DNA complexes, in which DNA interacts with crystalline Dps phases, L. lactis DNA:Dps complexes appeared as non‐crystalline aggregates of protein and DNA in electron micrographs.
Modulated targeting of GFP‐AtMAP65‐1 to central spindle microtubules during division
Authors: Mao Guojie , Chan Jordi , Calder Grant , Doonan John H. , Lloyd Clive W.
Published: July 2005

AtMAP65‐1 bundles cortical microtubules and we examined how this property is regulated during division in time‐lapse studies of Arabidopsis suspension cells expressing GFP‐AtMAP65‐1. Spindle fluorescence is diffuse during metaphase, restored to the central spindle at anaphase and then compacted at the midline during late anaphase/early telophase. However, mutagenesis of the microtubule‐associated protein (MAP) consensus Cdk site to a non‐phosphorylatable form allows premature decoration of microtubules traversing the central region of the metaphase spindle without affecting the timing of the subsequent compaction. This suggests that mutagenesis does not affect compaction but does affect a phosphorylation/dephosphorylation switch that normally targets AtMAP65‐1 to the central spindle at the metaphase/anaphase transition. GFP‐AtMAP65‐1 continues to label the midline of the early phragmoplast, suggesting a structural continuity with the central spindle – both structures being composed of anti‐parallel microtubules. However, once the cytokinetic apparatus expands into a ring the MAP becomes depleted at the midline. Despite this, cytokinesis is not arrested and membrane and callose are deposited at the cell plate. It is concluded that AtMAP65‐1 plays a role in the central spindle at anaphase to early cytokinesis but is not essential at the midline of the phragmoplast at later stages.
Tiling array‐driven elucidation of transcriptional structures based on maximum‐likelihood and Markov models
Authors: Toyoda Tetsuro , Shinozaki Kazuo
Published: July 2005

Tiling arrays of high‐density oligonucleotide probes spanning the entire genome are powerful tools for the discovery of new genes. However, it is difficult to determine the structure of the spliced product of a structurally unknown gene from noisy array signals only. Here we introduce a statistical method that estimates the precise splicing points and the exon/intron structure of a structurally unknown gene by maximizing the odds or the ratio of posterior probabilities of the structure under the observation of array signal intensities and nucleic acid sequences. Our method more accurately predicted the gene structures than the simple threshold‐based method, and more correctly estimated the expression values of structurally unknown genes than the window‐based method. It was observed that the Markov model contributed to the precision of splice points, and that the statistical significance of expression (P‐value) represented the reliability of the estimated gene structure and expression value well. We have implemented the method as a program ARTADE (ARabidopsis Tiling Array‐based Detection of Exons) and applied it to the Arabidopsis thaliana whole‐genome array data analysis. The database of the predicted results and the ARTADE program are available at http://omicspace.riken.jp/ARTADE/.
UPF3 suppresses aberrant spliced mRNA in Arabidopsis
Authors: Hori Koichi , Watanabe Yuichiro
Published: July 2005

It has been reported that eukaryotic organisms have a nonsense‐mediated mRNA decay (NMD) system to exclude aberrant mRNAs that produce truncated proteins. NMD is an RNA surveillance pathway that degrades mRNAs possessing premature translation termination codons (PTCs), thus avoiding production of possibly toxic truncated proteins. Three interacting proteins, UPF1, UPF2 and UPF3, are required for NMD in mammals and yeasts, and their amino acid sequences are well conserved among most eukaryotes, including plants. In this study, ‘The Arabidopsis Information Resource’ database was searched for mRNAs with premature termination codons. We selected five of these mRNAs and checked for the presence of PTCs in these mRNAs when translated in vivo. As a result we identified aberrant mRNAs produced by alternative splicing for each gene. These genes produced at least one alternative splicing variant including a PTC (PTC+) and another variant without a PTC (PTC−). We analyzed their PTC+/PTC− ratios in wild‐type Arabidopsis and upf3 mutant plants and showed that the PTC+/PTC− ratios were higher in atupf3 mutant plants than wild‐type plants and that the atupf3 mutant was less able to degrade mRNAs with premature termination codons than wild‐type plants. This indicated that the AtUPF3 gene is required by the plant NMD system to obviate aberrantly spliced mRNA.
Identification, subcellular localization and biochemical characterization of water‐soluble heteroglycans (SHG) in leaves of Arabidopsis thaliana L.: distinct SHG reside in the cytosol and in the apoplast
Authors: Fettke Joerg , Eckermann Nora , Tiessen Axel , Geigenberger Peter , Steup Martin
Published: July 2005

Water‐soluble heteroglycans (SHG) were isolated from leaves of wild‐type Arabidopsis thaliana L. and from two starch‐deficient mutants. Major constituents of the SHG are arabinose, galactose, rhamnose, and glucose. SHG was separated into low (<10 kDa; SHGS) and high (>10 kDa; SHGL) molecular weight compounds. SHGS was resolved into approximately 25 distinct oligoglycans by ion exchange chromatography. SHGL was further separated into two subfractions, designated as subfraction I and II, by field flow fractionation. For the intracellular localization of the various SHG compounds several approaches were chosen: first, leaf material was subjected to non‐aqueous fractionation. The apolar gradient fractions were characterized by monitoring markers and were used as starting material for the SHG isolation. Subfraction I and SHGS exhibited a distribution similar to that of cytosolic markers whereas subfraction II cofractionated with crystalline cellulose. Secondly, intact organelles were isolated and used for SHG isolation. Preparations of intact organelles (mitochondria plus peroxisomes) contained no significant amount of any heteroglycan. In isolated intact microsomes a series of oligoglycans was recovered but neither subfraction I nor II. In in vitro assays using glucose 1‐phosphate and recombinant cytosolic (Pho 2) phosphorylase both SHGS and subfraction I acted as glucosyl acceptor whereas subfraction II was essentially inactive. Rabbit muscle phosphorylase a did not utilize any of the plant glycans indicating a specific Pho 2–glycan interaction. As revealed by in vivo labeling experiments using 14CO2 carbon fluxes into subfraction I and II differed. Furthermore, in leaves the pool size of subfraction I varied during the light–dark regime.
A new role of the Arabidopsis SEPALLATA3 gene revealed by its constitutive expression
Authors: Castillejo Cristina , Romera‐Branchat Maida , Pelaz Soraya
Published: July 2005

During Arabidopsis flower development a set of homeotic genes plays a central role in specifying the distinct floral organs of the four whorls, sepals in the outermost whorl, and petals, stamens, and carpels in the sequentially inner whorls. The current model for the identity of the floral organs includes the SEPALLATA genes that act in combination with the A, B and C genes for the specification of sepals, petals, stamens and carpels. According to this new model, the floral organ identity proteins would form different complexes of proteins for the activation of the downstream genes. We show that the presence of SEPALLATA proteins is needed to activate the AG downstream gene SHATTERPROOF2, and that SEPALLATA4 alone does not provide with enough SEPALLATA activity for the complex to be functional. Our results suggest that CAULIFLOWER may be part of the protein complex responsible for petal development and that it is fully required in the absence of APETALA1 in 35S::SEP3 plants. In addition, genetic and molecular experiments using plants constitutively expressing SEPALLATA3 revealed a new role of SEPALLATA3 in activating other B and C function genes. We molecularly prove that the ectopic expression of SEPALLATA3 is sufficient to ectopically activate APETALA3 and AGAMOUS. Remarkably, plants that constitutively express both SEPALLATA3 and LEAFY developed ectopic petals, carpels and ovules outside of the floral context.
Identification of QTLs for domestication‐related and agronomic traits in an Oryza sativa×O. rufipogon BC1F7 population
Authors: Lee S.‐J. , Oh C.‐S. , Suh J.‐P. , McCouch S. R. , Ahn S.‐N.
Published: July 2005

Asian cultivated rice was domesticated from the wild rice, Oryza rufipogon and throughout the domestication process, a wide range of morphological and physiological changes altered the ancestral form. This study was conducted to identify the genetic basis of changes associated with the domestication process. An recombinant Inbred line (RIL) population consisting of 120 lines was developed from a cross between the Juponica cultivar.‘Hwayeongbyeo’and a presumed wild progenitor. O. rufipogon Griff. Acc.01944. The population was genotyped with 124 simple sequence length repeat (SSR) markers, providing an average interval size of 15 cM, and also evaluated for 20 traits related to domestication and agricultural performance. A total of 63 quantitative trait locus (QTLs) and one locus associated with qualitative variation for pericarp coloration were identified using single point and composite interval analysis. The number of QTLs per trait ranged from one to seven. Phenotypic variation associated with each QTL ranged from 3.7 to 40.4%. with an average of 15.3%. The results indicated that most domestication‐related traits clustered in chromosomal blocks, and the positions of many of these clusters were consistent with those reported in previous studies and with skewed segregation ratios in these BC1,F7 RILs. For 13 (20.6%) of the QTLs identified in this study. the O. rufipogan ‐derived allele contributed a desirable agronomic effect despite the overall undesirable characteristics of the wild phenotype. Favourable alleles from O. rufipogan were detected for panicle length, spikelets per panicle, days to heading and leaf discoloration associated with cold stress. When compared with previous studies involving interspecific crosses, it can be concluded that O. rufipogon is useful as a source of valuable alleles for rice improvement and that many of the introgressed regions contain genes that have a favourable impact on phenotype in different genetic backgrounds and different environments.
Changes in the meiotic pairing behaviour of a winter wheat‐winter barley hybrid maintained for a long term in tissue culture, and tracing the barley chromatin in the progeny using GISH and SSR markers
Authors: Molnár‐Lánci M. , Novotny C. , Linc G. , Naoy E. D.
Published: July 2005

The aim of the present study was to produce backcross progenies in a new winter wheat (‘Asakaze komugi’) × winter barley (‘Manas’) hybrid produced in Martonvasar. As no backcross seeds were obtained from the initial hybrids, young inflorescences of the hybrids were used for in vitro multiplication in three consecutive cycles until a backcross progeny was developed. The chromosome constitution of the regenerated hybrids was analysed using genomic in situ hybridization (GISH) after each in vitro multiplication cycle. The seven barley chromosomes were present even after the third in vitro multiplication cycle but abnormalities were observed. Sixteen BC; plants containing, according to GfSH analysis, one to three complete barley chromosomes, two deletion barley chromosomes and a dicentric wheat‐barley translocation were grown to maturity from the single backcross progeny. The barley chromatin was identified using 20 chromosome‐specific barley SSR markers. All seven barley chromosomes were represented in the BC: plants. A deletion breakpoint at FL ±0,3 on the 5HL chromosome arm facilitated the physical localization of microsatellite markers.
The clinical features of respiratory syncytial virus: Lower respiratory tract infection after upper respiratory tract infection due to influenza virus
Authors: Nishimura Nozomi , Nishio Hisahide , Lee Myeong Jin , Uemura Kanjiro
Published: July 2005

Background
: Respiratory syncytial virus (RSV) and influenza virus are the primary pathogens of respiratory tract infection. However, epidemics of influenza virus infection have been observed to interrupt RSV epidemics (termed an epidemiological interference effect).

Methods
: At a clinic in Tsuna county, Hyogo prefecture, Japan, a total of 1262 outpatients under 6 years of age with lower respiratory tract infection due to RSV (RSV‐LRTI) and upper respiratory tract infection due to influenza virus (FLU‐URTI) in three successive winter seasons (1999–2000, 2000–2001 and 2001–2002) were analyzed.

Results
: The RSV‐LRTI epidemic and FLU‐URTI epidemic overlapped in each season, but the RSV‐LRTI epidemic peak preceded that of the FLU‐URTI epidemic. Epidemiological interference between RSV and influenza virus was observed in the second and third season; the number of patients with RSV‐LRTI began to decrease after the start of the FLU‐URTI epidemic and recovered to some extent after the FLU‐URTI epidemic passed its peak. There were no differences in onset age, male‐to‐female ratio and severity of RSV‐LRTI in outpatients before and after the start of the FLU‐URTI epidemic in all the three seasons.

Conclusion
: An epidemiological interference between RSV and influenza virus was observed in Tsuna county in two of the three winter seasons. However, there was no difference between the clinical features of the patients with RSV‐LRTI before and after the start of the influenza virus infection epidemic. The data suggest that the clinical severity of RSV infection is not changed by the epidemiological interference effect of influenza virus infection epidemics.