bluuebunny/crossref_metadata_2025 huggingface dataset:
Combined targeting of lentiviral vectors and positioning of transduced cells by magnetic nanoparticles
Authors: Hofmann Andreas , Wenzel Daniela , Becher Ulrich M. , Freitag Daniel F. , Klein Alexandra M. , Eberbeck Dietmar , Schulte Maike , Zimmermann Katrin , Bergemann Christian , Gleich Bernhard , Roell Wilhelm , Weyh Thomas , Trahms Lutz , Nickenig Georg , Fleischmann Bernd K. , Pfeifer Alexander
Published: January 2009

Targeting of viral vectors is a major challenge for in vivo gene delivery, especially after intravascular application. In addition, targeting of the endothelium itself would be of importance for gene-based therapies of vascular disease. Here, we used magnetic nanoparticles (MNPs) to combine cell transduction and positioning in the vascular system under clinically relevant, nonpermissive conditions, including hydrodynamic forces and hypothermia. The use of MNPs enhanced transduction efficiency of endothelial cells and enabled direct endothelial targeting of lentiviral vectors (LVs) by magnetic force, even in perfused vessels. In addition, application of external magnetic fields to mice significantly changed LV/MNP biodistribution in vivo. LV/MNP-transduced cells exhibited superparamagnetic behavior as measured by magnetorelaxometry, and they were efficiently retained by magnetic fields. The magnetic interactions were strong enough to position MNP-containing endothelial cells at the intima of vessels under physiological flow conditions. Importantly, magnetic positioning of MNP-labeled cells was also achieved in vivo in an injury model of the mouse carotid artery. Intravascular gene targeting can be combined with positioning of the transduced cells via nanomagnetic particles, thereby combining gene- and cell-based therapies.
Localizing interference during naming: Convergent neuroimaging and neuropsychological evidence for the function of Broca's area
Authors: Schnur Tatiana T. , Schwartz Myrna F. , Kimberg Daniel Y. , Hirshorn Elizabeth , Coslett H. Branch , Thompson-Schill Sharon L.
Published: January 2009

To produce a word, the intended word must be selected from a competing set of other words. In other domains where competition affects the selection process, the left inferior frontal gyrus (LIFG) responds to competition among incompatible representations. The aim of this study was to test whether the LIFG is necessary for resolution of competition in word production. Using a methodological approach applying the same rigorous analytic methods to neuropsychological data as is done with neuroimaging data, we compared brain activation patterns in normal speakers (using fMRI) with the results of lesion-deficit correlations in aphasic speakers who performed the same word production task designed to elicit competition during lexical selection. The degree of activation of the LIFG in normal speakers and damage to the LIFG in aphasic speakers was associated with performance on the production task. These convergent findings provide strong support for the hypothesis that the region of cortex commonly known as Broca's area (i.e., the posterior LIFG) serves to bias competitive interactions during language production.
ChIP-on-chip significance analysis reveals large-scale binding and regulation by human transcription factor oncogenes
Authors: Margolin Adam A. , Palomero Teresa , Sumazin Pavel , Califano Andrea , Ferrando Adolfo A. , Stolovitzky Gustavo
Published: January 2009

ChIP-on-chip has emerged as a powerful tool to dissect the complex network of regulatory interactions between transcription factors and their targets. However, most ChIP-on-chip analysis methods use conservative approaches aimed at minimizing false-positive transcription factor targets. We present a model with improved sensitivity in detecting binding events from ChIP-on-chip data. Its application to human T cells, followed by extensive biochemical validation, reveals that 3 oncogenic transcription factors, NOTCH1, MYC, and HES1, bind to several thousand target gene promoters, up to an order of magnitude increase over conventional analysis methods. Gene expression profiling upon NOTCH1 inhibition shows broad-scale functional regulation across the entire range of predicted target genes, establishing a closer link between occupancy and regulation. Finally, the increased sensitivity reveals a combinatorial regulatory program in which MYC cobinds to virtually all NOTCH1-bound promoters. Overall, these results suggest an unappreciated complexity of transcriptional regulatory networks and highlight the fundamental importance of genome-scale analysis to represent transcriptional programs.
Continuous elimination of oxidized nucleotides is necessary to prevent rapid onset of cellular senescence
Authors: Rai Priyamvada , Onder Tamer T. , Young Jennifer J. , McFaline Jose L. , Pang Bo , Dedon Peter C. , Weinberg Robert A.
Published: January 2009

Reactive oxygen species (ROS) appear to play a role in limiting both cellular and organismic lifespan. However, because of their pleiotropic effects, it has been difficult to ascribe a specific role to ROS in initiating the process of cellular senescence. We have studied the effects of oxidative DNA damage on cell proliferation, believing that such damage is of central importance to triggering senescence. To do so, we devised a strategy to decouple levels of 8-oxoguanine, a major oxidative DNA lesion, from ROS levels. Suppression of MTH1 expression, which hydrolyzes 8-oxo-dGTP, was accompanied by increased total cellular 8-oxoguanine levels and caused early-passage primary and telomerase-immortalized human skin fibroblasts to rapidly undergo senescence, doing so without altering cellular ROS levels. This senescent phenotype recapitulated several salient features of replicative senescence, notably the presence of senescence-associated beta-galactosidase (SA beta-gal) activity, apparently irreparable genomic DNA breaks, and elevation of p21
Cip1
, p53, and p16
INK4A
tumor suppressor protein levels. Culturing cells under low oxygen tension (3%) largely prevented the shMTH1-dependent senescent phenotype. These results indicate that the nucleotide pool is a critical target of intracellular ROS and that oxidized nucleotides, unless continuously eliminated, can rapidly induce cell senescence through signaling pathways very similar to those activated during replicative senescence.
Genetic inactivation of D-amino acid oxidase enhances extinction and reversal learning in mice
Authors: Labrie Viviane , Duffy Steven , Wang Wei , Barger Steven W. , Baker Glen B. , Roder John C.
Published: January 2009

Activation of theN-methyl-d-aspartate receptor (NMDAR) glycine site has been shown to accelerate adaptive forms of learning that may benefit psychopathologies involving cognitive and perseverative disturbances. In this study, the effects of increasing the brain levels of the endogenous NMDAR glycine site agonist D-serine, through the genetic inactivation of its catabolic enzyme D-amino acid oxidase (DAO), were examined in behavioral tests of learning and memory. In the Morris water maze task (MWM), mice carrying the hypofunctionalDao1G181Rmutation demonstrated normal acquisition of a single platform location but had substantially improved memory for a new target location in the subsequent reversal phase. Furthermore,Dao1G181Rmutant animals exhibited an increased rate of extinction in the MWM that was similarly observed following pharmacological administration of D-serine (600 mg/kg) in wild-type C57BL/6J mice. In contextual and cued fear conditioning, no alterations were found in initial associative memory recall; however, extinction of the contextual fear memory was facilitated in mutant animals. Thus, an augmented level of D-serine resulting from reduced DAO activity promotes adaptive learning in response to changing conditions. The NMDAR glycine site and DAO may be promising therapeutic targets to improve cognitive flexibility and inhibitory learning in psychiatric disorders such as schizophrenia and anxiety syndromes.
Critical role of the cholinergic system for object-in-place associative recognition memory
Authors: Barker Gareth R.I. , Warburton Elizabeth C.
Published: January 2009

Object-in-place memory, which relies on the formation of associations between an object and the place in which it was encountered, depends upon a neural circuit comprising the perirhinal (PRH) and medial prefrontal (mPFC) cortices. This study examined the contribution of muscarinic cholinergic neurotransmission within this circuit to such object-in-place associative memory. Intracerebral administration of scopolamine in the PRH or mPFC impaired memory acquisition, but not retrieval and importantly we showed that unilateral blockade of muscarinic receptors simultaneously in both regions in opposite hemispheres, significantly impaired performance. Thus, object-in-place associative memory depends upon cholinergic modulation of neurones within the PRH-PFC circuit.
Role of dopamine receptors subtypes, D1-like and D2-like, within the nucleus accumbens subregions, core and shell, on memory consolidation in the one-trial inhibitory avoidance task
Authors: Managò Francesca , Castellano Claudio , Oliverio Alberto , Mele Andrea , De Leonibus Elvira
Published: January 2009

Recent evidence demonstrated that dopamine within the nucleus accumbens mediates consolidation of both associative and nonassociative memories. However, the specific contribution of the nucleus accumbens subregions, core and shell, and of D1 and D2 receptors subtypes has not been yet clarified. The aim of this study was, therefore, to directly compare the effect of D1 and D2 dopamine receptor blockade within the core and the shell subregions of the nucleus accumbens on memory consolidation. Using the one-trial inhibitory avoidance task in CD1 mice, we demonstrated that SCH 23390 (vehicle, 12.5, 25, 50 ng/side) administration within the core, but not the shell, impaired step-through latency 24 h after the administration if injected immediately, but not 120 min post-training. Interestingly, sulpiride (vehicle, 25, 50 ng/side) injection in both the core and the shell of the accumbens affected step-through latency 24 h later; also, in this case the impairment was time dependent. These data provide the most complete and direct demonstration to date that early consolidation of aversive memory requires D2 receptor activation in both nucleus accumbens subregions, and D1 activation selectively in the nucleus accumbens core.
Social modulation of associative fear learning by pheromone communication
Authors: Bredy Timothy W. , Barad Mark
Published: January 2009

Mice communicate through visual, vocal, and olfactory cues that influence innate, nonassociative behavior. We here report that exposure to a recently fear-conditioned familiar mouse impairs acquisition of conditioned fear and facilitates fear extinction, effects mimicked by both an olfactory chemosignal emitted by a recently fear-conditioned familiar mouse and by the putative stress-related anxiogenic pheromone β-phenylethylamine (β-PEA). Together, these findings suggest social modulation of higher-order cognitive processing through pheromone communication and support the concurrent excitor hypothesis of extinction learning.
Highly Diversified Molecular Evolution of Downstream Transcription Start Sites in Rice and Arabidopsis
Authors: Tanaka Tsuyoshi , Koyanagi Kanako O. , Itoh Takeshi
Published: January 2009

Alternative usage of transcription start sites (TSSs) is one of the key mechanisms to generate gene variation in eukaryotes. Here, we show diversified molecular evolution of TSSs in remotely related flowering plants, rice (Oryza sativa) and Arabidopsis (Arabidopsis thaliana), by comprehensive analyses of large collections of full-length cDNAs and genome sequences. We determined 45,917 representative TSSs within 23,445 loci of rice and 35,313 TSSs within 16,964 loci of Arabidopsis, about two TSSs per locus in either species. The nucleotide features around TSSs displayed distinct patterns when the most upstream TSSs were compared with downstream TSSs. We found that CG-skew and AT-skew were clearly different between upstream and downstream TSSs, and that this difference was commonly observed in rice and Arabidopsis. Relative entropy analysis revealed that the most upstream TSSs had retained canonical cis elements, whereas downstream TSSs showed atypical nucleotide features. Expression patterns were distinguishable between upstream and downstream TSSs. These results indicate that plant TSSs were generally diversified in downstream regions, resulting in the development of new gene expression patterns. Furthermore, our comparative analysis of TSS variation between the species showed a positive correlation between TSS number and gene conservation. Rice and Arabidopsis might have evolved novel TSSs in an independent manner, which led to diversification of these two species.
An Integrated Genomics Approach to Define Niche Establishment byRhodococcus fascians
Authors: Depuydt Stephen , Trenkamp Sandra , Fernie Alisdair R. , Elftieh Samira , Renou Jean-Pierre , Vuylsteke Marnik , Holsters Marcelle , Vereecke Danny
Published: January 2009

Rhodococcus fascians is a Gram-positive phytopathogen that induces shooty hyperplasia on its hosts through the secretion of cytokinins. Global transcriptomics using microarrays combined with profiling of primary metabolites on infected Arabidopsis (Arabidopsis thaliana) plants revealed that this actinomycete modulated pathways to convert its host into a niche. The transcript data demonstrated that R. fascians leaves a very characteristic mark on Arabidopsis with a pronounced cytokinin response illustrated by the activation of cytokinin perception, signal transduction, and homeostasis. The microarray data further suggested active suppression of an oxidative burst during the R. fascians pathology, and comparison with publicly available transcript data sets implied a central role for auxin in the prevention of plant defense activation. Gene Ontology categorization of the differentially expressed genes hinted at a significant impact of infection on the primary metabolism of the host, which was confirmed by subsequent metabolite profiling. The much higher levels of sugars and amino acids in infected plants are presumably accessed by the bacteria as carbon and nitrogen sources to support epiphytic and endophytic colonization. Hexoses, accumulating from a significantly increased invertase activity, possibly inhibited the expression of photosynthesis genes and photosynthetic activity in infected leaves. Altogether, these changes are indicative of sink development in symptomatic tissues. The metabolomics data furthermore point to the possible occurrence of secondary signaling during the interaction, which might contribute to symptom development. These data are placed in the context of regulation of bacterial virulence gene expression, suppression of defense, infection phenotype, and niche establishment.
How Does Cyclic Electron Flow Alleviate Photoinhibition in Arabidopsis?
Authors: Takahashi Shunichi , Milward Sara E. , Fan Da-Yong , Chow Wah Soon , Badger Murray R.
Published: January 2009

Cyclic electron flow (CEF) around photosystem I has a role in avoiding photoinhibition of photosystem II (PSII), which occurs under conditions in which the rate of photodamage to PSII exceeds the rate of its repair. However, the molecular mechanism underlying how CEF contributes to photoprotection is not yet well understood. We examined the effect of impairment of CEF and thermal energy dissipation (qE) on photoinhibition using CEF (pgr5) and qE (npq1 and npq4) mutants of Arabidopsis (Arabidopsis thaliana) exposed to strong light. Impairment of CEF by mutation of pgr5 suppressed qE and accelerated photoinhibition. We found that impairment of qE, by mutations of pgr5, npq1, and npq4, caused inhibition of the repair of photodamaged PSII at the step of the de novo synthesis of the D1 protein. In the presence of the chloroplast protein synthesis inhibitor chloramphenicol, impairment of CEF, but not impairment of qE, accelerated photoinhibition, and a similar effect was obtained when leaves were infiltrated with the protonophore nigericin. These results suggest that CEF-dependent generation of ΔpH across the thylakoid membrane helps to alleviate photoinhibition by at least two different photoprotection mechanisms: one is linked to qE generation and prevents the inhibition of the repair of photodamaged PSII at the step of protein synthesis, and the other is independent of qE and suppresses photodamage to PSII.
Phthoxazolin A inhibits prostate cancer growth by modulating tumor–stromal cell interactions
Authors: Kawada Manabu , Inoue Hiroyuki , Usami Ihomi , Ikeda Daishiro
Published: January 2009

Because stroma in tumor tissues can promote prostate cancer development, modulation of tumor–stromal cell interactions may represent an attractive new strategy for cancer treatment. Here, we report that phthoxazolin A and its analog inthomycin B inhibit the growth of human prostate cancer DU‐145 cells by modulating tumor–stromal cell interactions. Using an in vitro coculture system, in which prostate cancer cell growth is upregulated by prostate stromal cells (PrSC), we found that phthoxazolin A and inthomycin B strongly inhibited the growth of DU‐145 cells when in coculture with PrSC compared to DU‐145 cells cultured alone. Although PrSC consist of both fibroblasts and myofibroblasts, phthoxazolin A and inthomycin B inhibited the expression of smooth muscle α‐actin, a myofibroblast marker, without affecting vimentin and β‐actin expression. Because myofibroblasts secrete various factors that can promote tumor cell growth, we examined whether the inhibitory compounds affected the secretion of such factors from PrSC. Proteomic analysis and reverse transcription–polymerase chain reaction revealed that phthoxazolin A and inthomycin B inhibited the expression of several insulin‐like growth factor binding proteins and insulin‐like growth factor (IGF)‐I by PrSC. Transforming growth factor‐β1 increased myofibroblast numbers and IGF‐I levels in PrSC. Phthoxazolin A inhibited transforming growth factor‐β1 activity without altering phosphorylation of the downstream molecule smad2. Furthermore, conditioned medium from phthoxazolin A‐treated PrSC failed to increase the phosphorylation of IGF‐IR and Akt in DU‐145 cells. Taken together, our results suggested that phthoxazolin A acts as a small‐molecule modulator of tumor–stromal cell interactions that can indirectly suppress prostate cancer cell growth through inhibition of IGF‐I production by PrSC. (Cancer Sci 2009; 100: 150–157)
Ligand‐independent higher‐order multimerization of CXCR4, a G‐protein‐coupled chemokine receptor involved in targeted metastasis
Authors: Hamatake Makiko , Aoki Toru , Futahashi Yuko , Urano Emiko , Yamamoto Naoki , Komano Jun
Published: January 2009

CXCR4, a G‐protein‐coupled receptor of CXCL12/stromal cell‐derived factor‐1α, mediates a wide range of physiological and pathological processes, including the targeted metastasis of cancer cells. CXCR4 has been shown to homo‐oligomerize in several experimental systems. However, it remains unclear with which domains CXCR4 interacts homotypically, and whether it dimerizes or forms a higher‐order complex. To address these issues, we used bioluminescent resonance energy transfer and bimolecular fluorescence complementation analyses to measure the homotypic interactions of CXCR4 in living cells. Both assays indicated that CXCR4 interacts homotypically, which is consistent with previous studies. By studying CXCR4 mutants lacking various domains, we found that multiple transmembrane domains probably serve as potential molecular interaction surfaces for oligomerization. The relative contribution of the amino‐ or carboxy‐termini to oligomerization was small. To differentiate between a dimer and a multimer consisting of more than two molecules, bioluminescent resonance energy transfer–bimolecular fluorescence complementation analysis was conducted. It revealed that CXCR4 engages in higher‐order oligomerization in a ligand‐independent fashion. This is the first report providing direct experimental evidence for the higher‐order multimerization of CXCR4 in vivo. We hypothesize that CXCR4 distributes to the cell surface as a multimer, in order to effectively sense, with increased avidity, the chemotaxis‐inducing ligand in the microenvironment. Studying the structure and function of the oligomeric state of CXCR4 may lead us to develop novel CXCR4 inhibitors that disassemble the molecular cluster of CXCR4. (Cancer Sci 2009; 100: 95–102)
Primary cutaneous T‐cell lymphoma of unspecified type with cytotoxic phenotype: Clinicopathological analysis of 27 patients
Authors: Hagiwara Masahiro , Takata Katsuyoshi , Shimoyama Yoshie , Yamamoto Kazuhito , Takahashi Emiko , Asano Naoko , Iwase Yuko , Okazaki Yoshiko , Tamada Yasuhiko , Yoshino Tadashi , Tomita Yasushi , Nakamura Shigeo
Published: January 2009

The objective of our study was to investigate the clinicopathological features of the currently ill‐defined subtype of primary cutaneous T‐cell lymphoma of unspecified type (CTCLU) with a cytotoxic phenotype and no Epstein–Barr virus (EBV) association. A series of 27 patients with CTCLU (median age 49 years; range 25–87 years; 18 men) was reviewed. Performance status scores above 1 (7%), clinical stages above 2 (15%), B symptoms (26%), extracutaneous involvement (30%), and a fatal course within 1 year of diagnosis (19%) were observed infrequently. The International Prognostic Index was high or high to intermediate in 11%, and the Prognostic Index for Peripheral T‐cell Lymphoma unspecified was above group 2 in 22%. Notably, the rates of spontaneous regression and T‐cell receptor gene rearrangements by polymerase chain reaction analysis were seen in 26 and 17% of our cases, respectively. Histologically, 22 patients had subcutaneous involvement of whom eight showed a lethal clinical course, and five patients without subcutaneous involvement were all survivors. Immunophenotypical and morphological features allowed us to subclassify our cases according to the following four categories: (1) epidermotropic CD8+ T‐cell lymphoma (n = 5); (2) cutaneous γ/δ T‐cell lymphoma (n = 8); (3) cutaneous α/β pleomorphic T‐cell lymphoma (n = 8); and (4) cutaneous medium/large pleomorphic T‐cell lymphoma, not otherwise specified (n = 6). All four of these groups of lymphomas exhibited a relatively favorable clinical course compared to previous reports. However, epidermotropic CD8+ T‐cell lymphoma appeared to be unique with a higher ratio (80%) of spontaneous regression, a lower ratio (40%) of subcutaneous involvement, and a more favorable clinical course than the other three subcategories. (Cancer Sci 2009; 100: 33–41)
Population pharmacokinetics of docetaxel in patients with hepatic dysfunction treated in an oncology practice
Authors: Minami Hironobu , Kawada Kenji , Sasaki Yasutsuna , Tahara Makoto , Igarashi Tadahiko , Itoh Kuniaki , Fujii Hirofumi , Saeki Toshiaki , Ozawa Kazuhiro , Sato Hitoshi
Published: January 2009

To investigate the relationship between the degree of liver dysfunction and the pharmacokinetics of docetaxel, a population pharmacokinetic model was developed in an oncology practice without excluding patients with moderate to severe liver dysfunction. Two hundred patients were treated with docetaxel as a single agent or in combination chemotherapy. The plasma concentration–time course data were analyzed using a three‐compartment open model with zero‐order administration and first‐order elimination on the NONMEM program. Sixty‐one had elevated transaminase levels, and alkaline phosphatase was elevated in 40. Body surface area, albumin, α1‐acid glycoprotein, and liver function were found to be significant covariates for the systemic clearance of docetaxel. Compared to patients with normal or minimal impairment of liver function, patients with grade 2 and 3 elevations of transaminases at baseline in conjunction with elevation of alkaline phosphatase had 22 and 38% lower clearances, respectively. Goodness‐of‐fit plots indicated that the model was fitted well with the observed data, and the bootstrap method guaranteed robustness of the model. We developed a population pharmacokinetic model for docetaxel, which can be used in the setting of an oncology practice. Based on the model, dose reduction by approximately 20 and 40% should be considered for patients with grade 2 and 3 elevations of transaminases at baseline in conjunction with elevation of alkaline phosphatase, respectively. (Cancer Sci 2009; 100: 144–149)
Identification of new type III effectors and analysis of the plant response by competitive index
Authors: MACHO ALBERTO P. , RUIZ‐ALBERT JAVIER , TORNERO PABLO , BEUZÓN CARMEN R.
Published: January 2009

In recent years, many efforts have been directed towards the identification of new type III‐secreted effectors, and the completion of the secretomes of several Pseudomonas syringae pathovars. Several functional and bioinformatic screenings have been used to search for candidates, which have been tested for translocation into the plant cell, an essential criterion for the identification of new type III effector proteins. The most common translocation assay is based on the use of ΔAvrRpt2 as a reporter. When fused to a type III effector protein, ΔAvrRpt2 is translocated and elicits a hypersensitive response in leaves of Arabidopsis thaliana expressing the RPS2 resistance protein. This approach has been used widely and has allowed the identification of a considerable number of new effectors in a fast and convenient manner. However, as the hypersensitive response is a semi‐quantitative assay, and the conditions do not resemble those occurring in nature, effectors with low expression or translocation efficiency could fail to translocate sufficient ΔAvrRpt2 to trigger a clear hypersensitive response. In keeping with these limitations, this test has failed to detect some true effectors that have been confirmed as such by other means. In order to increase the sensitivity of this method, we have developed a modification of the ΔAvrRpt2‐based translocation assay using a competitive index in mixed infection to monitor the limitation of growth associated with the induction of the hypersensitive response. We have tested several effector candidates from P. syringae pv. phaseolicola and other P. syringae pathovars, and have compared the results obtained by our competitive index translocation assay with those obtained by standard hypersensitive response assays. We have identified six type III secretion system‐translocated proteins using this approach, five of which failed to be identified by hypersensitive response assays. In addition, we have analysed the defence response triggered by one of these effectors using competitive index assays.
Identification of virulence genes in Fusarium oxysporum f. sp. lycopersici by large‐scale transposon tagging
Authors: LÓPEZ‐BERGES MANUEL SÁNCHEZ , DI PIETRO ANTONIO , DABOUSSI MARIE‐JOSÉE , WAHAB HALA ABDEL , VASNIER CHRISTELLE , RONCERO M. ISABEL G. , DUFRESNE MARIE , HERA CONCEPCIÓN
Published: January 2009

Forward genetic screens are efficient tools for the dissection of complex biological processes, such as fungal pathogenicity. A transposon tagging system was developed in the vascular wilt fungus Fusarium oxysporum f. sp. lycopersici by inserting the novel modified impala element imp160::gfp upstream of the Aspergillus nidulans niaD gene, followed by transactivation with a constitutively expressed transposase. A collection of 2072 Nia+ revertants was obtained from reporter strain T12 and screened for alterations in virulence, using a rapid assay for invasive growth on apple slices. Seven strains exhibited reduced virulence on both apple slices and intact tomato plants. Five of these were true revertants showing the re‐insertion of imp160::gfp within or upstream of predicted coding regions, whereas the other two showed either excision without re‐insertion or no excision. Linkage between imp160::gfp insertion and virulence phenotype was determined in four transposon‐tagged loci using targeted deletion in the wild‐type strain. Knockout mutants in one of the genes, FOXG_00016, displayed significantly reduced virulence, and complementation of the original revertant with the wild‐type FOXG_00016 allele fully restored virulence. FOXG_00016 has homology to the velvet gene family of A. nidulans. The high rate of untagged virulence mutations in the T12 reporter strain appears to be associated with increased genetic instability, possibly as a result of the transactivation of endogenous transposable elements by the constitutively expressed transposase.
CD4+CD25+FoxP3+ regulatory T cells are increased whilst CD3+CD4−CD8−αβTCR+ Double Negative T cells are decreased in the peripheral blood of patients with multiple myeloma which correlates with disease burden
Authors: Feyler Sylvia , Von Lilienfeld‐Toal Marie , Jarmin Sarah , Marles Lee , Rawstron Andy , Ashcroft A. J. , Owen Roger G. , Selby Peter J. , Cook Gordon
Published: January 2009

Increased levels of naturally occurring regulatory T cells (TReg cells) have been found in a variety of solid tumours and haematological malignancies. In multiple myeloma (MM), evidence suggests that TReg cells are increased though controversy exists with regards to their function and no relationship to disease stage and treatment has been demonstrated. Here, we demonstrate significantly elevated levels of functional CD4+CD25+FoxP3+ TReg cells in a large cohort of patients with MM as well as monoclonal gammopathy of uncertain significance (MGUS) in comparison to age‐matched, healthy controls. The frequency of Double Negative TReg cells was also evaluated, demonstrating that these cells were reduced in patients with MM. Furthermore, a characteristic profile of immunomodulatory cytokines in the peripheral blood and bone marrow of patients with MM and MGUS was demonstrated, compared with healthy controls. This data adds further evidence to the understanding of the role of TReg cell subsets in tumour immunology and the fundamentals of the host/tumour immune conflict.
Soybean β‐conglycinin as the main allergen in a patient with food‐dependent exercise‐induced anaphylaxis by tofu: food processing alters pepsin resistance
Authors: Adachi A. , Horikawa T. , Shimizu H. , Sarayama Y. , Ogawa T. , Sjolander S. , Tanaka A. , Moriyama T.
Published: January 2009

Background
Food‐dependent exercise‐induced anaphylaxis (FDEIA) due to soybeans is a rare disorder. The allergen responsible for FDEIA due to soybeans has not yet been determined.

Objective
We characterized the clinical features of a patient with FDEIA due to tofu, who was well tolerant to drinking soy milk. We then sought to identify the responsible soybean allergen(s) in that patient. We further studied whether different stabilities of the allergen(s) to pepsin digestion between two soybean products are related to their clinical allergenicity.

Methods
Skin prick tests and provocation tests using soybean products were performed to detect the responsible food and other factors that induced the allergic symptoms. Specific IgE to various soybean allergens were examined by ImmunoCAP®, ELISA and protein microarray assays. Immunoblotting for soybeans and soybean products using the patient's serum was also performed. Soybean products were serially digested by pepsin to disclose the stability of the allergens.

Results
Provocation with ingestion of tofu and exercise induced the allergic symptoms, while ingestion of soy milk and exercise did not. Immunoblot analysis, ELISA and protein microarray assay revealed that β‐conglycinin mainly reacts with IgE antibodies in the patient's serum. By immunoblot analysis, β‐conglycinin in soy milk completely disappeared after pepsin digestion within 20 min, whereas β‐conglycinin in tofu was almost intact after more than 120 min of pepsin digestion.

Conclusion
We identified β‐conglycinin as the causative allergen in a patient with FDEIA induced by tofu. The difference in resistance to pepsin digestion between tofu and soy milk suggests that the presence of undigested allergens in the digestive tract is a prerequisite for the development of FDEIA.
Implementation of pre‐seasonal sublingual immunotherapy with a five‐grass pollen tablet during optimal dosage assessment
Authors: Horak F. , Jaeger S. , Worm M. , Melac M. , Didier A.
Published: January 2009

Background
The optimal dose (300IR) of a five‐grass pollen sublingual immunotherapy tablet in terms of efficacy was previously demonstrated from the first pollen season.

Objective
Here, we aim to confirm whether this dose remained optimal during the peak of the pollen season by assessing the efficacy and quality of life data.

Methods
A total of 628 subjects with grass pollen rhinoconjunctivitis were randomized in a double‐blind, placebo‐controlled, multi‐centre, pan‐European trial. Subjects received once‐daily tablets (Stallergenes, Antony, France) of 100IR, 300IR, 500IR or placebo, starting 4 months before and throughout the 2005 grass pollen season. The pollen season was defined as the first day of 3 consecutive days with a grass pollen count above 30 grains/m3 of air, recorded using Hirst‐type volumetric pollen traps, to the last day before 3 consecutive days with a pollen count below 30 grains/m3.

Results
The grass pollen season lasted an average of 30 days, with a peak of 12 days. The mean treatment duration before the grass pollen season was similar in the four treatment groups (121.4±31.1 to 128.6±15.4 days in the safety population). Both the 300IR and 500IR groups had highly significant improvements in Rhinoconjunctivitis Total Symptom Score (RTSS) vs. placebo at the peak pollen season (P=0.0005 and 0.0014, respectively), which agreed with improvements in RTSS in the primary evaluations. The average RTSS scores were slightly elevated during the peak pollen season in all treatment groups. The overall Rhinoconjunctivitis Quality of Life Questionnaire score confirmed the optimal dosage 300IR at peak (P<0.0001) and at the end (P0.0031) of the pollen season. All doses were well tolerated.

Conclusion
At the peak pollen season, the efficacy and quality of life data for both 300IR and 500IR groups was significantly improved vs. the placebo group. These results confirm the conclusions of the primary evaluations and validate the use of 300IR tablets for clinical practice.
Services in the community for adults with psychosis and intellectual disabilities: a Delphi consultation of professionals' views
Authors: Hemmings C. P. , Underwood L. A. , Bouras N.
Published: January 2009

Background  There remains a severe lack of evidence on the effectiveness of community services for adults with psychosis and intellectual disabilities (ID). There has been little consensus even of what services should provide for this service user group.

Method  A consultation of multidisciplinary professionals was carried out by using a three‐round Delphi exercise. Participants were recruited nationally. They rated their views on the importance of 139 items for the care of adults with psychosis and ID. These included 85 routine service components, 23 service user characteristics for those needing a more intensive service and 31 more intensive service components.

Results  Forty‐nine out of 52 participants completed all three rounds of the Delphi consultation. Consensus of opinion (≥80% agreement as essential) was obtained on 18 of the routine service components, nine of the service user characteristics and five of the more intensive service components. The routine service components considered essential can be broadly considered under a need for a focused approach on the service user and their illness (e.g. monitoring of mental state) and the added need to work within the wider context of the service user with psychosis and ID (e.g. access to social, leisure or occupational activities). Five of the more intensive service components were considered to be essential (e.g. can react to a crisis that day). However, the routine service components considered essential already contained many components such as out‐of‐hours support and crisis plans also relevant to more intensive services.

Conclusion  These findings can be used to develop further the evidence base for services in the community for this user group and to assist in the preparation of much needed service evaluation studies.
Programmed drug‐dependent ribosome stalling
Authors: Ramu Haripriya , Mankin Alexander , Vazquez‐Laslop Nora
Published: January 2009

The ribosome has the intrinsic capacity to monitor the sequence and structure of the nascent peptide. This fundamental property of the ribosome is often exploited in regulation of gene expression, in particular, for activation of expression of genes conferring resistance to ribosome‐targeting antibiotics. Induction of expression of these genes is controlled by the programmed stalling of the ribosome at a regulatory open reading frame located upstream of the resistance cistron. Formation of the stalled translation complex depends on the presence of an antibiotic in the ribosome exit tunnel and the sequence of the nascent peptide. In this review, we summarize our current understanding of the molecular mechanisms of drug‐ and nascent peptide‐dependent ribosome stalling.
What is plant behaviour?*
Authors: TREWAVAS ANTHONY
Published: January 2009

The nature of plant behaviour is discussed, and it is concluded that it is best described as what plants do. The possibility that plant behaviour is simply signal‐induced phenotypic plasticity is outlined, and some limitations of this assumption are considered. Natural environments present many challenges to growing plants, and the consequent signalling that plants perceive is becoming extremely complex. Plant behaviour is active, purposeful and intentional, and examples are discussed. Much plant behaviour, concerned with stress and herbivory, is also based on an assessment of the future likelihood of further damaging episodes and is therefore predictive. Plant behaviour involves the acquisition and processing of information. Informational terminology provides a suitable way of incorporating the concepts of learning, memory and intelligence into plant behaviour, capabilities that plants are rarely credited with. Finally, trade‐offs, cost–benefit assessments and decision making are common plant behavioural attributes. It is suggested that intelligent assessments that involve the whole plant are essential to optimize these adaptive capabilities.
Cell‐Mediated Immune Responses and Protective Efficacy against Infection with Mycobacterium tuberculosis Induced by Hsp65 and hIL‐2 Fusion Protein in Mice
Authors: Shi C. , Yuan S. , Zhang H. , Zhang T. , Wang L. , Xu Z.
Published: January 2009

Heat shock protein 65 (Hsp65) is an important immunodominant antigen against tuberculosis (TB), and interleukin‐2 (IL‐2) plays an important role in the regulation of antimycobacteria immune responses. In order to further increase the immunogenicity of Hsp65 against infection caused by Mycobacterium tuberculosis (MTB), we expressed MTB Hsp65 and human IL‐2 fusion protein, Hsp65‐hIL‐2, in Escherichia coli. The expression of Hsp65‐hIL‐2 was confirmed by Western blotting using anti‐Hsp65 MoAb and anti‐hIL‐2 MoAb, respectively. Hsp65‐IL‐2 and Hsp65 were then purified by Ni‐NTA affinity chromatography. Mice were immunized with purified Hsp65‐hIL‐2 or Hsp65 emulsified in the adjuvant combination dimethyl dioctadecylammonium bromide and monophosphoryl lipid A. Eight weeks after immunization, there was significant proliferation of spleen lymphocytes in response to both Hsp65 and Hsp65‐hIL‐2 proteins. Interestingly, Hsp65‐hIL‐2 fusion protein elicited significantly higher levels of IFN‐γ and IL‐2 in the lymphocytes culture supernatant than that of the BCG (Denmark strain) immunized group and Hsp65 group (P < 0.05). After challenging the immunized mice with MTB, the bacteria loads in the spleens and lungs of mice immunized with the fusion protein were significantly lower than Hsp65 alone group, reaching an equivalent level as BCG immunization group. Our results suggest that the Hsp65 and hIL‐2 fusion protein may serve as an alternative vaccine candidate against MTB infection.
Renal transplant donation from a deceased cardiac graft recipient: A case for marginal donors
Authors: Guzzo Isabella , Capozza Nicola , Parisi Francesco , Collura Giuseppe , Diomedi Camassei Francesca , Dello Strologo Luca
Published: January 2009

Abstract:  We describe a case of dual kidney transplant in a highly sensitized 18‐yr‐old male patient followed for chronic renal failure secondary to focal and segmental glomerulosclerosis. The donor was a six‐yr‐old boy who had received a cardiac transplant at the age of three months and had been treated with CNIs for over five yr. Renal biopsy performed before the transplant revealed focal sclerosis in 30–40% of the donor’s glomeruli. Considering the long expected time on the waiting list for this highly sensitized boy and the absence of prohibited antigens in the donor, we decided to perform the transplant. To increase renal mass, both kidneys were transplanted. Clinical course was good but the patient developed proteinuria, which improved with ramipril. We performed two renal biopsies in the first nine months post‐transplant which showed a progressive increase of the glomerulosclerosis. Despite the histology, renal function remains good 24 months after transplant. This is the first report of a transplant using a kidney from a donor who had received long‐term CNI treatment. Short‐term outcome is satisfactory. We suggest that marginal donors could also be considered for donation in pediatric age for selected groups of patients.
LE RECOURS À LA THÉRAPEUTIQUE ÉLECTRIQUE DANS LE TRAITEMENT D'URGENCE DES TROUBLES AFFECTANT LE RYTHME ET LA FUNCTION CARDIAQUE
Authors: Manni C. , Reale A.
Published: January 2009

Les auteurs relatent leur expéerience lorsqu'il s'est agi de traiter les troubles du rythme qui peuvent exiger un traitement d'urgence: fibrillation auriculaire très fréquente, tachycardie auriculaire paroxystique, tachycardie et fibrillation ventriculaires, blocs atrio‐ventriculaires et arrět du coeur. A leur avis, si les conditions ambiantes pepermettent, chacune des complications ci‐dessus conseille nettement la thérapeutique électrique, sans que l'on s'attarde à des tentatives de traitement pharmacologique.

Les auteurs étayent leur opinion sur l'utilité de cette pratique en rapportant certains cas qu'ils ont pu observer lors d'examens hémodynamiques et pendant des interventions de cardiochirurgie. Selon les circonstances, on a utilisé des défibrillateurs externes ou internes, avec ou sans synchronisation, avec ou sans stimulation, à l'aide d'électrodes endocardiques ou myocardiques.

La stimulation par impulsions couplées, moyennant une amplification postextra‐systolique, a donné des résultats marquants, měme si la méthode doit ětre considérée encore dans la phase de l'expérimentation clinique.
DIE DRINGLICHE BEHANDLUNG DER AKUTEN STORUNGENDES HERZRHYTMUS NACH CHIRURGISCHEN EINGRIFFEN IN DER BRUSTHOHLE
Authors: Lebedewa R. N.
Published: January 2009

Die Behandlung des Vorhofflimmerns‐ und ‐Flatterm, das sich nach thorakalen Operationen entwickelt, wurde bei 180 Kranken und die Behandlung der Ventrikelfibrillation ‐ bei 17 durchgefiihrt. Fur die Behandlung des Vorhofflimmerns‐ und ‐Flatterns wurden Chinidin, Novokain‐amid, Cocarboxilase und Kontraschock benutzt. Bei der Anwendung von Chinidin und Novokainamid wurde ein positives Ergebnk bei 87,7 yo erreicht, bei der Anwendung des Kontraschock bei 95 yo der Kranken. Bei der Wiederherstellung des Sinusrhythmus sind Faille, von akuten Thromboembolie nicht aufgetreten. 2 Kranke hatten durch Chinidinbehandlung einen Herzstillstand, den es gelang zu bewaltigen. Bei den Kranken, die mit Kontraschock behandelt wurden, stellte man in 1/3 der Faille die Wiederherstellung von Vorhofflimmern fest. Die Behandlung der Ventrikelfibrillation war weniger wirksam, weil sie sich gewohnlich auf der Basis schwerer postoperativer Komplikationen entwickelte. Von 17 Fallen gelang es den Sinusrhythmus nur bei 9 Kranken wiederherzustellen, 3 Kranke uberlebten.
LA CONDUITE DES OXYDASES GRANULOCYTAIRES DANS LA NEUROLEPTANALGESIE
Authors: Emanuelli H. , Ventafridda V. , Talamazzi F. , Testa S. , Prada A.
Published: January 2009

RESUME

Les Auteurs ont étudié la conduite des oxydases granulocytaires (cytochromeoxydase) avec une méthode de coloration des éléments du sang (nadirreaktion modifiée) sur des échantillons de sang périphérique obtenus à intervalle déterminé avant, pendant et après la neuroleptanalgésie.

Les résultats obtenus ont démontré une conduite anormale de ces éléments, signe d'une utilisation altérée de l'O2 au niveau moléculaire.
CONTRIBUTIONS AU CONTROLE DE L'ANESTHESIE PAR LA MESURE DE LA RESISTIVITE DU TEGUMENT ET DU pH DE LA SECRETION SUDORALE
Authors: Popa C. Eugene
Published: January 2009

La mesure de l'impédance du tégument pendant l'anesthésie, comme test quantitatif, associé, à l'appréciation du pH de la sécrétion sudorale comme test qualitatif, sont des moyens simples et utiles pour le contrǒle de l'analgésie et le maintien d'une protection néurovégétative équilibrée et convenable. L'intensité et la durée de la vasoconstriction dans le territoire cutané est proportionelle à l'augmentation de l'acidité de la transpiration et de la conductibilité électrique du tégument. La sécretion sudorale est indépendante de la vascularisation.

Les tests‐associés é la surveillance du pouls et de la tension artérielle, gardent leur valeur dans l'analgésie neuroleptique qui peut ětre maintenue à un niveau sélectif, avec la prédominance analgétique sur la neurolepsie.
Ketamine® (Cl‐581) in Clinical Anaesthesia
Authors: Gjessing Jon
Published: January 2009

The observations made in the 40 patients show that Ketamine® (C1‐581) analgesia is satisfactory for many kinds of surgery in old people and even suggest that results improve with increasing age. C1‐581 was least satisfactory in major abdominal surgery, in which the need for complete relaxation necessitated supplementary anaesthesia in more than half the cases.

Neither circulation nor respiration was depressed, which is of great importance in the age group treated, but a high incidence of subjective experiences occurred in the awakening stage.

SCHLUSSFOLGERUNGEN UND ZUSAMMENFASSUNG

Die bei 40 Patienten gemachten Beobachtungen zeigten, dass die mit Ketamin (C1‐581) erzielte Analgesie für viele Arten von Eingriffen bei alten Leuten zufriedenstellend ist. Die Resultate erscheinen sogar mit zunehmenden Alter besser. Am wenigsten zufriedenstellend erwies sich C1‐581 in der grossen Ab‐dominalchirurgie wegen der für komplette Erschaffung notwendigen zusätzlichen Anaesthesie bei mehr als der Hälfte der Fälle.

Weder Kreislauf noch Atmung waren gedämpft, was in der behandelten Altersgruppe von Bedeutung war, hingegen hatten die Patienten im Aufwach‐stadium sehr häufig subjektive Sensationen.
CONTROLLED RESPIRATION IN THE MANAGEMENT OF PATIENTS WITH TRAUMATIC BRAIN INJURIES
Authors: Gordon Emeric
Published: January 2009

The clinical records of 252 patients with traumatic brain lesions admitted unconscious to hospital were studied. Of the 252 patients, 51 were treated with artificial hyperventilation. The final clinical outcome in these two groups are compared. The recovery rate was significantly higher and the mortality rate significantiy lower in the group of patients treated with hyperventilation. The possible reasons for these differences are discussed, and it is concluded diat patients with severe traumatic brain injuries benefit from an adequate period of treatment widi artificial hyperventilation.

Die Krankengeschichten von 252 Patienten, die bewusstlos mit traumatischen Gehirnschäden zur Aufnahme gekommen waren, wurden einem eingehenden Studium unterzogen. Von diesen 252 Patienten waren 51 mit künstlicher Hyperventilation behandelt worden. Das klinische Endresultat dieser beiden Gruppen wird verglichen. Bei den mit Hyperventilation behandelten Kranken war die Erholungsrate signifikant höher und die Mortalität signifikant niedriger. Die Erklärungsmöglichkeiten für diesen Unterschied werden diskutiert, wobei sich die Schlussfolgerung aufdrängt, dass für Patienten mit schweren traumatischen Gehirnläsionen eine adäquate Behandlungsperiode mit künsdicher Hyperventilation vorteilhaft ist.