bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytoplasmic dynein nomenclature
Authors: Pfister K. Kevin , Fisher Elizabeth M.C. , Gibbons Ian R. , Hays Thomas S. , Holzbaur Erika L.F. , McIntosh J. Richard , Porter Mary E. , Schroer Trina A. , Vaughan Kevin T. , Witman George B. , King Stephen M. , Vallee Richard B.
Published: November 2005

A variety of names has been used in the literature for the subunits of cytoplasmic dynein complexes. Thus, there is a strong need for a more definitive consensus statement on nomenclature. This is especially important for mammalian cytoplasmic dyneins, many subunits of which are encoded by multiple genes. We propose names for the mammalian cytoplasmic dynein subunit genes and proteins that reflect the phylogenetic relationships of the genes and the published studies clarifying the functions of the polypeptides. This nomenclature recognizes the two distinct cytoplasmic dynein complexes and has the flexibility to accommodate the discovery of new subunits and isoforms.
RNA-associated autoantigens activate B cells by combined B cell antigen receptor/Toll-like receptor 7 engagement
Authors: Lau Christina M. , Broughton Courtney , Tabor Abigail S. , Akira Shizuo , Flavell Richard A. , Mamula Mark J. , Christensen Sean R. , Shlomchik Mark J. , Viglianti Gregory A. , Rifkin Ian R. , Marshak-Rothstein Ann
Published: November 2005

Previous studies (Leadbetter, E.A., I.R. Rifkin, A.H. Hohlbaum, B. Beaudette, M.J. Shlomchik, and A. Marshak-Rothstein. 2002. Nature. 416:603–607; Viglianti, G.A., C.M. Lau, T.M. Hanley, B.A. Miko, M.J. Shlomchik, and A. Marshak-Rothstein. 2003. Immunity. 19:837–847) established the unique capacity of DNA and DNA-associated autoantigens to activate autoreactive B cells via sequential engagement of the B cell antigen receptor (BCR) and Toll-like receptor (TLR) 9. We demonstrate that this two-receptor paradigm can be extended to the BCR/TLR7 activation of autoreactive B cells by RNA and RNA-associated autoantigens. These data implicate TLR recognition of endogenous ligands in the response to both DNA- and RNA-associated autoantigens. Importantly, the response to RNA-associated autoantigens was markedly enhanced by IFN-α, a cytokine strongly linked to disease progression in patients with systemic lupus erythematosus (SLE). As further evidence that TLRs play a key role in autoantibody responses in SLE, we found that autoimmune-prone mice, lacking the TLR adaptor protein MyD88, had markedly reduced chromatin, Sm, and rheumatoid factor autoantibody titers.
Characterization of marginal zone B cell precursors
Authors: Srivastava Bhaskar , Quinn William J. , Hazard Kristin , Erikson Jan , Allman David
Published: November 2005

Selection of recently formed B cells into the follicular or marginal zone (MZ) compartments is proposed to occur by way of proliferative intermediates expressing high levels of CD21/35 and CD23. However, we show that CD21/35high CD23+ splenocytes are not enriched for proliferative cells, and do not contribute substantially to the generation of follicular B cells. Instead, ontogenic relationships, steady-state labeling kinetics, and adoptive transfer experiments suggest that CD21/35high CD23+ splenocytes serve primarily as precursors for MZ B cells, although their developmental potential seems to be broader and is influenced by environmental cues that are associated with lymphopenia. Furthermore, CD21/35high CD23+ splenocytes share several key functional characteristics with MZ B cells, including their capacity to trap T-independent antigen and a heightened proliferative response to LPS. These observations challenge previous models of peripheral B cell maturation, and suggest that MZ B cells develop by way of CD21/35high CD23+ intermediates.
The insulin A-chain epitope recognized by human T cells is posttranslationally modified
Authors: Mannering Stuart I. , Harrison Leonard C. , Williamson Nicholas A. , Morris Jessica S. , Thearle Daniel J. , Jensen Kent P. , Kay Thomas W.H. , Rossjohn Jamie , Falk Ben A. , Nepom Gerald T. , Purcell Anthony W.
Published: November 2005

The autoimmune process that destroys the insulin-producing pancreatic β cells in type 1 diabetes (T1D) is targeted at insulin and its precursor, proinsulin. T cells that recognize the proximal A-chain of human insulin were identified recently in the pancreatic lymph nodes of subjects who had T1D. To investigate the specificity of proinsulin-specific T cells in T1D, we isolated human CD4+ T cell clones to proinsulin from the blood of a donor who had T1D. The clones recognized a naturally processed, HLA DR4–restricted epitope within the first 13 amino acids of the A-chain (A1–13) of human insulin. T cell recognition was dependent on the formation of a vicinal disulfide bond between adjacent cysteine residues at A6 and A7, which did not alter binding of the peptide to HLA DR4. CD4+ T cell clones that recognized this epitope were isolated from an HLA DR4+ child with autoantibodies to insulin, and therefore, at risk for T1D, but not from two healthy HLA DR4+ donors. We define for the first time a novel posttranslational modification that is required for T cell recognition of the insulin A-chain in T1D.
Live attenuated yellow fever 17D infects human DCs and allows for presentation of endogenous and recombinant T cell epitopes
Authors: Barba-Spaeth Giovanna , Longman Randy S. , Albert Matthew L. , Rice Charles M.
Published: November 2005

The yellow fever (YF) 17D vaccine is one of the most successful live attenuated vaccines available. A single immunization induces both long-lasting neutralizing antibody and YF-specific T cell responses. Surprisingly, the mechanism for this robust immunity has not been addressed. In light of several recent reports suggesting flavivirus interaction with dendritic cells (DCs), we investigated the mechanism of YF17D interaction with DCs and the importance of this interaction in generating T cell immunity. Our results show that YF17D can infect immature and mature human DCs. Viral entry is Ca2+ dependent, but it is independent of DC-SIGN as well as multiple integrins expressed on the DC surface. Similar to infection of cell lines, YF infection of immature DCs is cytopathic. Although infection itself does not induce DC maturation in vitro, TNF-α–induced maturation protects DCs from YF-induced cytopathogenicity. Furthermore, we show that DCs infected with YF17D or YF17D carrying a recombinant epitope can process and present antigens for CD8+ T cell stimulation. These findings offer insight into the immunologic mechanisms associated with the highly capable YF17D vaccine that may guide effective vaccine design.
Charge Screening by Internal pH and Polyvalent Cations as a Mechanism for Activation, Inhibition, and Rundown of TRPM7/MIC Channels
Authors: Kozak J. Ashot , Matsushita Masayuki , Nairn Angus C. , Cahalan Michael D.
Published: November 2005

The Mg2+-inhibited cation (MIC) current, believed to represent activity of TRPM7 channels, is found in lymphocytes and mast cells, cardiac and smooth muscle, and several other eukaryotic cell types. MIC current is activated during whole-cell dialysis with divalent-free internal solutions. Millimolar concentrations of intracellular Mg2+ (or other divalent metal cations) inhibit the channels in a voltage-independent manner. The nature of divalent inhibition and the mechanism of channel activation in an intact cell remain unknown. We show that the polyamines (spermine, spermidine, and putrescine) inhibit the MIC current, also in a voltage-independent manner, with a potency that parallels the number of charges. Neomycin and poly-lysine also potently inhibited MIC current in the absence of Mg2+. These same positively charged ions inhibited IRK1 current in parallel with MIC current, suggesting that they probably act by screening the head group phosphates on PIP2 and other membrane phospholipids. In agreement with this hypothesis, internal protons also inhibited MIC current. By contrast, tetramethylammonium, tetraethylammonium, and hexamethonium produced voltage-dependent block but no inhibition. We show that inhibition by internal polyvalent cations can be relieved by alkalinizing the cytosol using externally applied ammonium or by increasing pH in inside-out patches. Furthermore, in perforated-patch and cell-attached recordings, when intracellular Mg2+ is not depleted, endogenous MIC or recombinant TRPM7 currents are activated by cytosolic alkalinization and inhibited by acidification; and they can be reactivated by PIP2 following rundown in inside-out patches. We propose that MIC (TRPM7) channels are regulated by a charge screening mechanism and may function as sensors of intracellular pH.
Elevated telomere-telomere recombination in WRN-deficient, telomere dysfunctional cells promotes escape from senescence and engagement of the ALT pathway
Authors: Laud Purnima R. , Multani Asha S. , Bailey Susan M. , Wu Ling , Ma Jin , Kingsley Charles , Lebel Michel , Pathak Sen , DePinho Ronald A. , Chang Sandy
Published: November 2005

Werner Syndrome (WS) is characterized by premature aging, genomic instability, and cancer. The combined impact of WRN helicase deficiency and limiting telomere reserves is central to disease pathogenesis. Here, we report that cells doubly deficient for telomerase and WRN helicase show chromosomal aberrations and elevated recombination rates between telomeres of sister chromatids. Somatic reconstitution of WRN function, but not a WRN helicase-deficient mutant, abolished telomere sister chromatid exchange (T-SCE), indicating that WRN normally represses T-SCEs. Elevated T-SCE was associated with greater immortalization potential and resultant tumors maintained telomeres via the alternative lengthening of telomere (ALT) pathway. We propose that the increased incidence of chromosomal instability and cancer in WS relates in part to aberrant recombinations between sister chromatids at telomeres, which facilitates the activation of ALT and engenders cancer-relevant chromosomal aberrations and tumor formation.
The HIR corepressor complex binds to nucleosomes generating a distinct protein/DNA complex resistant to remodeling by SWI/SNF
Authors: Prochasson Philippe , Florens Laurence , Swanson Selene K. , Washburn Michael P. , Workman Jerry L.
Published: November 2005

The histone regulatory (HIR) and histone promoter control (HPC) repressor proteins regulate three of the four histone gene loci during the Saccharomyces cerevisiae cell cycle. Here, we demonstrate that Hir1, Hir2, Hir3, and Hpc2 proteins form a stable HIR repressor complex. The HIR complex promotes histone deposition onto DNA in vitro and constitutes a novel nucleosome assembly complex. The HIR complex stably binds to DNA and nucleosomes. Furthermore, HIR complex binding to nucleosomes forms a distinct protein/DNA complex resistant to remodeling by SWI/SNF. Thus, the HIR complex is a novel nucleosome assembly complex which functions with SWI/SNF to regulate transcription.
Lost in translation: the influence of ribosomes on bacterial mRNA decay: Figure 1.
Authors: Deana Atilio , Belasco Joel G.
Published: November 2005

The lifetimes of bacterial mRNAs are strongly affected by their association with ribosomes. Events occurring at any stage during translation, including ribosome binding, polypeptide elongation, or translation termination, can influence the susceptibility of mRNA to ribonuclease attack. Ribosomes usually act as protective barriers that impede mRNA cleavage, but in some instances they can instead trigger the decay of the mRNA to which they are bound or send a signal that leads to widespread mRNA destabilization within a cell. The influence of translation on mRNA decay provides a quality-control mechanism for minimizing the use of poorly or improperly translated mRNAs as templates for the production of abnormal proteins that might be toxic to bacteria.
Loss of splicing factor ASF/SF2 induces G2 cell cycle arrest and apoptosis, but inhibits internucleosomal DNA fragmentation
Authors: Li Xialu , Wang Jin , Manley James L.
Published: November 2005

ASF/SF2 is an SR protein splicing factor that participates in constitutive and alternative pre-mRNA splicing and is essential for cell viability. Using a genetically modified chicken B-cell line, DT40-ASF, we now show that ASF/SF2 inactivation results in a G2-phase cell cycle arrest and subsequent programmed cell death. However, although several hallmarks of apoptosis are apparent, internucleosomal DNA fragmentation was not detected. Furthermore, inactivation of ASF/SF2 also blocks DNA fragmentation normally induced by a variety of apoptotic stimuli. Notably, mRNA encoding the inhibitor of caspase-activated DNase-L (ICAD-L), which acts as an inhibitor as well as a chaperone of caspase-activated DNase (CAD), decreased in abundance, whereas the level of mRNA encoding ICAD-S, which has only inhibitory activity, increased upon ASF/SF2 depletion. Strikingly, expression of appropriate levels of exogenous human ICAD-L restored apoptotic DNA laddering in ASF/SF2-depleted cells. These results not only indicate that loss of an SR protein splicing factor can induce cell cycle arrest and apoptosis, but also illustrate the important role of ICAD and its regulation by alternative splicing in the process of apoptotic DNA fragmentation.
TAK1, but not TAB1 or TAB2, plays an essential role in multiple signaling pathways in vivo
Authors: Shim Jae-Hyuck , Xiao Changchun , Paschal Amber E. , Bailey Shannon T. , Rao Ping , Hayden Matthew S. , Lee Ki-Young , Bussey Crystal , Steckel Michael , Tanaka Nobuyuki , Yamada Gen , Akira Shizuo , Matsumoto Kunihiro , Ghosh Sankar
Published: November 2005

TGF-β-activated kinase 1 (TAK1), a member of the MAPKKK family, is thought to be a key modulator of the inducible transcription factors NF-κB and AP-1 and, therefore, plays a crucial role in regulating the genes that mediate inflammation. Although in vitro biochemical studies have revealed the existence of a TAK1 complex, which includes TAK1 and the adapter proteins TAB1 and TAB2, it remains unclear which members of this complex are essential for signaling. To analyze the function of TAK1 in vivo, we have deleted theTak1gene in mice, with the resulting phenotype being early embryonic lethality. Using embryonic fibroblasts lacking TAK1, TAB1, or TAB2, we have found that TNFR1, IL-1R, TLR3, and TLR4-mediated NF-κB and AP-1 activation are severely impaired inTak1m/mcells, but they are normal inTab1-/-andTab2-/-cells. In addition,Tak1m/mcells are highly sensitive to TNF-induced apoptosis. TAK1 mediates IKK activation in TNF-α and IL-1 signaling pathways, where it functions downstream of RIP1–TRAF2 and MyD88–IRAK1–TRAF6, respectively. However, TAK1 is not required for NF-κB activation through the alternative pathway following LT-β signaling. In the TGF-β signaling pathway, TAK1 deletion leads to impaired NF-κB and c-Jun N-terminal kinase (JNK) activation without impacting Smad2 activation or TGF-β-induced gene expression. Therefore, our studies suggests that TAK1 acts as an upstream activating kinase for IKKβ and JNK, but not IKKα, revealing an unexpectedly specific role of TAK1 in inflammatory signaling pathways.
The tumor suppressorsInk4candp53collaborate independently withPatchedto suppress medulloblastoma formation
Authors: Uziel Tamar , Zindy Frederique , Xie Suqing , Lee Youngsoo , Forget Antoine , Magdaleno Susan , Rehg Jerold E. , Calabrese Christopher , Solecki David , Eberhart Charles G. , Sherr Sarah E. , Plimmer Sarah , Clifford Steven C. , Hatten Mary E. , McKinnon Peter J. , Gilbertson Richard J. , Curran Tom , Sherr Charles J. , Roussel Martine F.
Published: November 2005

Recurrent genetic alterations in human medulloblastoma (MB) include mutations in the sonic hedgehog (SHH) signaling pathway andTP53inactivation (∼25% and 10% of cases, respectively). However, mouse models of MB, regardless of their initiating lesions, generally depend uponp53inactivation for rapid onset and high penetrance. The gene encoding the cyclin-dependent kinase inhibitor p18Ink4cis transiently expressed in mouse cerebellar granule neuronal precursor cells (GNPs) as they exit the cell division cycle and differentiate. Coinactivation ofInk4candp53provided cultured GNPs with an additive proliferative advantage, either in the presence or absence of Shh, and induced MB with low penetrance but with greatly increased incidence following postnatal irradiation. In contrast, mice lacking one or two functionalInk4calleles and one copy ofPatched(Ptc1) encoding the Shh receptor rapidly developed MBs that retained wild-typep53. In tumor cells purified from double heterozygotes, the wild-typePtc1allele, but notInk4c, was inactivated. Therefore, when combined withPtc1mutation,Ink4cis haploinsufficient for tumor suppression. Methylation ofINK4C(CDKN2C) was observed in four of 23 human MBs, and p18INK4Cprotein expression was extinguished in 14 of 73 cases. Hence, p18INK4Closs may contribute to MB formation in children.
The PAS/LOV protein VIVID supports a rapidly dampened daytime oscillator that facilitates entrainment of the Neurospora circadian clock
Authors: Elvin Mark , Loros Jennifer J. , Dunlap Jay C. , Heintzen Christian
Published: November 2005

A light-entrainable circadian clock controls development and physiology in Neurospora crassa. Existing simple models for resetting based on light pulses (so-called nonparametric entrainment) predict that constant light should quickly send the clock to an arrhythmic state; however, such a clock would be of little use to an organism in changing photoperiods in the wild, and we confirm that true, albeit dampened, rhythmicity can be observed in extended light. This rhythmicity requires the PAS/LOV protein VIVID (VVD) that acts, in the light, to facilitate expression of an oscillator that is related to, but distinguishable from, the classic FREQUENCY/WHITE-COLLAR complex (FRQ/WCC)-based oscillator that runs in darkness. VVD prevents light resetting of the clock at dawn but, by influencing frq RNA turnover, promotes resetting at dusk, thereby allowing the clock to run through the dawn transition and take its phase cues from dusk. Consistent with this, loss of VVD yields a clock whose performance follows the simple predictions of earlier models, and overexpression of VVD restores rhythmicity in the light and sensitivity of phase to the duration of the photoperiod.
Solution structure of γS‐crystallin by molecular fragment replacement NMR
Authors: Wu Zhengrong , Delaglio Frank , Wyatt Keith , Wistow Graeme , Bax Ad
Published: November 2005

The solution structure of murine γS‐crystallin (γS) has been determined by multidimensional triple resonance NMR spectroscopy, using restraints derived from two sets of dipolar couplings, recorded in different alignment media, and supplemented by a small number of NOE distance restraints. γS consists of two topologically similar domains, arranged with an approximate twofold symmetry, and each domain shows close structural homology to closely related (∼50% sequence identity) domains found in other members of the γ‐crystallin family. Each domain consists of two four‐strand “Greek key” β‐sheets. Although the domains are tightly anchored to one another by the hydrophobic surfaces of the two inner Greek key motifs, the N‐arm, the interdomain linker and several turn regions show unexpected flexibility and disorder in solution. This may contribute entropic stabilization to the protein in solution, but may also indicate nucleation sites for unfolding or other structural transitions. The method used for solving the γS structure relies on the recently introduced molecular fragment replacement method, which capitalizes on the large database of protein structures previously solved by X‐ray crystallography and NMR.
Modulation of oxygen binding to insect hemoglobins: The structure of hemoglobin from the botfly Gasterophilus intestinalis
Authors: Pesce Alessandra , Nardini Marco , Dewilde Sylvia , Hoogewijs David , Ascenzi Paolo , Moens Luc , Bolognesi Martino
Published: November 2005

Hemoglobins (Hbs) reversibly bind gaseous diatomic ligands (e.g., O2) as the sixth heme axial ligand of the penta‐coordinate deoxygenated form. Selected members of the Hb superfamily, however, display a functionally relevant hexa‐coordinate heme Fe atom in their deoxygenated state. Endogenous heme hexa‐coordination is generally provided in these Hbs by the E7 residue (often His), which thus modulates accessibility to the heme distal pocket and reactivity of the heme toward exogenous ligands. Such a pivotal role of the E7 residue is prominently shown by analysis of the functional and structural properties of insect Hbs. Here, we report the 2.6 Å crystal structure of oxygenated Gasterophilus intestinalis Hb1, a Hb known to display a penta‐coordinate heme in the deoxygenated form. The structure is analyzed in comparison with those of Drosophila melanogaster Hb, exhibiting a hexa‐coordinate heme in its deoxygenated derivative, and of Chironomus thummi thummi HbIII, which displays a penta‐coordinate heme in the deoxygenated form. Despite evident structural differences in the heme distal pockets, the distinct molecular mechanisms regulating O2 binding to the three insect Hbs result in similar O2 affinities (P50 values ranging between 0.12 torr and 0.46 torr).
Crystal structures of T cell receptor β chains related to rheumatoid arthritis
Authors: Li Hongmin , Van Vranken Sandra , Zhao Yiwei , Li Zhong , Guo Yi , Eisele Leslie , Li Yixin
Published: November 2005

The crystal structures of the Vβ17+ β chains of two human T cell receptors (TCRs), originally derived from the synovial fluid (SF4) and tissue (C5–1) of a patient with rheumatoid arthritis (RA), have been determined in native (SF4) and mutant (C5–1F104→Y/C187→S) forms, respectively. These TCR β chains form homo‐dimers in solution and in crystals. Structural comparison reveals that the main‐chain conformations in the CDR regions of the C5–1 and SF4 Vβ17 closely resemble those of a Vβ17 JM22 in a bound form; however, the CDR3 region shows different conformations among these three Vβ17 structures. At the side‐chain level, conformational differences were observed at the CDR2 regions between our two ligand‐free forms and the bound JM22 form. Other significant differences were observed at the Vβ regions 8–12, 40–44, and 82–88 between C5–1/SF4 and JM22 Vβ17, implying that there is considerable variability in the structures of very similar β chains. Structural alignments also reveal a considerable variation in the Vβ–Cβ associations, and this may affect ligand recognition. The crystal structures also provide insights into the structure basis of T cell recognition of Mycoplasma arthritidis mitogen (MAM), a superantigen that may be implicated in the development of human RA. Structural comparisons of the Vβ domains of known TCR structures indicate that there are significant similarities among Vβ regions that are MAM‐reactive, whereas there appear to be significant structural differences among those Vβ regions that lack MAM‐reactivity. It further reveals that CDR2 and framework region (FR) 3 are likely to account for the binding of TCR to MAM.
The crystal structure of a novel SAM‐dependent methyltransferase PH1915 from Pyrococcus horikoshii
Authors: Sun Warren , Xu Xiaohui , Pavlova Marina , Edwards Aled M. , Joachimiak Andrzej , Savchenko Alexei , Christendat Dinesh
Published: November 2005

The S‐adenosyl‐L‐methionine (SAM)‐dependent methyltransferases represent a diverse and biologically important class of enzymes. These enzymes utilize the ubiquitous methyl donor SAM as a cofactor to methylate proteins, small molecules, lipids, and nucleic acids. Here we present the crystal structure of PH1915 from Pyrococcus horikoshii OT3, a predicted SAM‐dependent methyltransferase. This protein belongs to the Cluster of Orthologous Group 1092, and the presented crystal structure is the first representative structure of this protein family. Based on sequence and 3D structure analysis, we have made valuable functional insights that will facilitate further studies for characterizing this group of proteins. Specifically, we propose that PH1915 and its orthologs are rRNA‐ or tRNA‐specific methyltransferases.
Interactions of microorganisms isolated from gilthead sea bream, Sparus aurata L., on Vibrio harveyi, a pathogen of farmed Senegalese sole, Solea senegalensis (Kaup)
Authors: Chabrillón M , Rico R M , Arijo S , Díaz‐Rosales P , Balebona M C , Moriñigo M A
Published: November 2005

Four bacterial isolates from farmed gilthead sea bream, Sparus aurata, included in a previous study as members of the Vibrionaceae and Pseudomonodaceae and the genus Micrococcus, have been evaluated for their adhesive ability to skin and intestinal mucus of farmed Senegalese sole, Solea senegalensis, and their antagonistic effect on Vibrio harveyi, a pathogen of sole. These isolates showed higher adhesion to sole mucus than the pathogenic strains of V. harveyi assayed. Only two of the isolates showed antagonistic activity to V. harveyi. Interactions of the four isolates with V. harveyi in respect of adhesion to skin and intestinal mucus under exclusion, competition and displacement conditions were studied. Three isolates were able to reduce the attachment to skin and intestinal sole mucus of a pathogenic strain of V. harveyi under displacement and exclusion conditions, but not under competition conditions. The in vivo probiotic potential of isolate Pdp11 was assessed by oral administration followed by challenge with the pathogenic V. harveyi strain Lg14/00. A group of 50 Senegalese sole received a commercial diet supplemented with 108 cfu g−1 of lyophilized Lg14/00 for 15 days. A second group of fish received a non‐supplemented commercial diet. After challenge the mortality of the fish receiving the diet supplemented with the potential probiotic isolate was significantly lower than that in the fish receiving the non‐supplemented commercial diet. This study has shown that the ability to interfere with attachment of pathogens, as well as the adhesion to host surfaces, are suitable criteria for selection of candidate probiotics for use in the culture of Senegalese sole.
Development of a rapid assay for the diagnosis of Myxobolus cerebralis in fish and oligochaetes using loop‐mediated isothermal amplification
Authors: El‐Matbouli M , Soliman H
Published: November 2005

A loop‐mediated isothermal amplification assay was developed for the rapid detection of Myxobolus cerebralis in both fish and oligochaete hosts. The assay was optimized to amplify parasitic DNA by incubation with Bst DNA polymerase and a set of six specially constructed primers at 65 °C for 60 min. The amplification products were detected visually using SYBR Green I dye which gave identical results to gel electrophoresis analysis. Parasite DNA was detected from infected oligochaetes, and from the anal fin, caudal fin, dorsal fin and operculum of clinically infected fish. This ‘Myxo‐LAMP’ assay has a detection limit similar to that of a polymerase chain reaction assay (10−6), but is more rapid and only requires a water bath for amplification and is therefore practical for simple and rapid diagnosis of infected tissue.
Exhaled nitric oxide and exercise‐induced bronchoconstriction in young male conscripts: association only in atopics
Authors: Rouhos A. , Ekroos H. , Karjalainen J. , Sarna S. , Sovijärvi A. R. A.
Published: November 2005

This study was conducted to evaluate how bronchial responsiveness to direct and indirect stimuli relate to nitric oxide producing airway inflammation, and whether the relationship differs between atopic and nonatopic patients with various degrees of bronchial hyperresponsiveness and airway inflammation in a group of otherwise homogenous young men.

We studied 181 consecutive non‐smoking steroid‐naive young male conscripts referred to military hospital because of respiratory symptoms suggesting asthma. Skin prick tests, spirometry, measurement of exhaled nitric oxide (FENO), and standardized airway challenges with histamine and exercise were performed.

128 patients were atopic. FENO was significantly higher in the atopic group, median 21.2 ppb, compared to 10.2 ppb in the nonatopic group. Still, 36% of all nonatopic patients had elevated FENO. Bronchial responsiveness to histamine (HIB) was similar in the two groups, but exercise‐induced bronchoconstriction (EIB) was stronger in atopics (P < 0.01). FENO associated significantly with atopy (P < 0.001), severity of EIB (P < 0.001) and HIB (P = 0.006) in multiple linear regression model. In separate regression models for atopic and nonatopic patients FENO associated with severity of EIB and HIB in atopic patients only. The results were similar when patients with confirmed diagnosis of asthma were analyzed separately.

Our results indicate that FENO significantly associates with EIB and HIB in atopic, but not in nonatopic steroid‐naïve patients with asthmatic symptoms. The finding suggests that in such atopic patients degree of airway hyperresponsiveness may reflect severity of airway inflammation. However, in nonatopic patients with similar symptoms other mechanisms of airway hyperresponsiveness may be more important.
Quality of asthma care: results from a community pharmacy based survey
Authors: Laforest L. , Van Ganse E. , Devouassoux G. , Chretin S. , Bauguil G. , Pacheco Y. , Chamba G.
Published: November 2005

Background:  Optimal control is a major objective of disease management of asthma. The aim of the present study was to provide descriptive data on disease management in asthma patients, including medical resource utilization.

Methods:  Asthma patients (age 18–50 years) were consecutively recruited in 348 dispensing pharmacies. They completed a questionnaire which collected data on personal characteristics, asthma management, including medical resource utilization, including asthma management. Asthma control was measured with the Asthma Control Test. Data from computerized pharmacy records of medications, dispensed before inclusion, were also collected.

Results:  In 1791 eligible patients, 1559 accepted to participate in the study (mean age = 36.5, 56.1% of females). During the previous 4 weeks, the asthma control was satisfactory for only 28% of the patients, despite extensive provision of anti‐inflammatory asthma control treatments (89%). Combinations of long acting beta agonists (LABA) and inhaled corticosteroids (ICS) were commonly used (59%), while fewer patients received LABA and ICS as two separate medications (15%). In addition, short‐acting beta agonists, were frequently dispensed (71%). A substantial number of patients consulted their GPs on a monthly basis. Patients commonly reported daily shortness of breath (30%), daily use of rescue medication (29%) and weekly nocturnal symptoms (32%). Surprisingly, most patients considered their asthma as completely or well controlled (76%).

Conclusions:  Our results clearly identify a need to improve the management of asthma. Education programmes would be beneficial to improve asthma control.
Are overweight asthmatics more difficult to control?
Authors: Saint‐Pierre P. , Bourdin A. , Chanez P. , Daures J.‐P. , Godard P.
Published: November 2005

The relationship between asthma and obesity appears to be quite complex. The aim of this study was to assess the effect of excess weight on asthma control evolution in a cohort of asthmatics. A prospective database was set up, which enrolled adult asthmatics with persistent (mild, moderate or severe) asthma. The control of asthma was defined as a binary variable, acceptable or unacceptable. In order to evaluate the effect of body mass index (BMI; <25 or ≥25), data were analysed using a continuous time homogeneous Markov model in which the forces ruling the transition between the two health states were estimated. The following confounding covariates were also evaluated in the model: severity of asthma, current treatment with oral corticosteroids (OCS) and history of OCS over the year preceding inclusion. About 406 asthmatics were included who made a total of 1639 consultations; the median length of follow up was 182 days. Using a univariate model, overweight patients had a lower risk of transiting from the unacceptable to the acceptable health state (RR = 0.45; P < 0.01). The effect of weight remained significant (RR = 0.53; P < 0.01) in the multivariate model including the other covariates. Moreover, transition probabilities stabilized more rapidly for patients with BMI < 25 (200 vs 300 days). In this study, we thus demonstrated that there is an association between excess weight and transition from unacceptable to acceptable control. Because control of asthma clearly drives asthma management, this finding has consequences for defining original new strategies for managing asthma in overweight patients.
Prevention and treatment of hymenoptera venom allergy: guidelines for clinical practice
Authors: Bonifazi F. , Jutel M. , Biló B. M. , Birnbaum J. , Muller U. ,   the EAACI Interest Group on Insect Venom Hypersensitivity
Published: November 2005

Based on the knowledge of the living conditions and habitat of social Aculeatae a series of recommendations have been formulated which can potentially greatly minimize the risk of field re‐sting. After a systemic sting reaction, patients should be referred to an allergy specialist for evaluation of their allergy, and if necessary venom immunotherapy (VIT). An emergency medical kit should be supplied, its use clearly demonstrated and repeatably practised until perfected. This should be done under the supervision of a doctor or a trained nurse. Epinephrine by intramuscolar injection is regarded as the treatment of choice for acute anaphylaxis. H1‐antihistamines alone or in combination with corticosteroids may be effective in mild to moderate reactions confined to the skin and may support the value of treatment with epinephrine in full‐blown anaphylaxis. Up to 75% of the patients with a history of systemic anaphylactic sting reaction develop systemic symptoms once again when re‐stung. Venom immunotherapy is a highly effective treatment for individuals with a history of systemic reaction and who have specific IgE to venom allergens. The efficacy of VIT in yellow jacket venom allergic patients has been demonstrated also by assessing health‐related quality of life. If both skin tests and serum venom specific IgE turn negative, VIT may be stopped after 3 years. After VIT lasting 3–5 years, most patients with mild to moderate anaphylactic symptoms remain protected following discontinuation of VIT even with positive skin tests. Longer term or lifelong treatment should be considered in high‐risk patients. Because of the small but relevant risk of re‐sting reactions, in these patients, emergency kits, including epinephrine auto‐injectors, should be discussed with every patient when stopping VIT.
Effects of acculturation and social exchange on the expectations of filial piety among Hispanic/Latino parents of adult children
Authors: Kao Hsueh‐Fen S. , Travis Shirley S.
Published: November 2005

Abstract  This study explores the effects of acculturation and social exchange variables on the expectations of filial piety (values and beliefs about parents’ care) among Hispanic/Latino parents of adult children. A convenience sample of 318 Hispanics/Latinos with adult children was assembled in Texas and North Carolina, USA, for face‐to‐face interviews in order to collect information about their families and to assess their expectations of filial piety by their adult children. The number of years in the USA, age, gender, the eldest child's age, and living with a person ≥  65 years of age had significant relationships to scores from a new Spanish version of the Expectations of Filial Piety Scale (EFPS). The significant variables explained 46.3% of the variance in the EFPS scores. A conceptual model that includes both acculturation and social exchange variables is a useful approach for understanding the expectations of filial piety by Hispanic/Latino parents of adult children.
Nurses’ attitudes and practices towards abused women
Authors: Häggblom A. M. E. , Hallberg L. R.‐M. , Möller Anders R.
Published: November 2005

Abstract  The present article is a descriptive survey assessing nurses’ knowledge, training, and practices regarding the care of abused women. A self‐administered questionnaire was sent to all nurses working within the government health organization on the Åland Islands, Finland. The response rate was 57%. Most nurses had no formal training in domestic violence and were less likely than the in‐service‐trained nurses to intervene with abused women. Nurses’ knowledge, beliefs, and practices were found to be unsystematic and had drawbacks. Nurses were not familiar with the formal structure provided by the health administration. The results suggest that in order to meet the urgent need for training among nurses, an in‐service training program needs to be set up. This program would help the nurses in identifying survivors of abuse and make them prepared to intervene more effectively to promote the health of these women.
Implementing clinical guidelines for nutrition in a neurosurgical intensive care unit
Authors: Annette Hansson , Wenström Yvonne
Published: November 2005

Abstract  Patients in neurosurgical intensive care have individual needs concerning nutrition because of their conditions. An important therapeutic goal is to prevent the development of malnutrition as it contributes to an increase in mortality and enhances the risk of complications in these patients. The health‐care team has a mutual responsibility for this, although it is a complex task and a correct assessment of patients’ nutritional needs is vital. Multidisciplinary clinical guidelines focused on nutrition might help the health‐care staff in decision‐making and allowing individualized treatment for patients. The aim of this study was to evaluate the implementation and use of such a guideline. The results show that the guideline is used in varying degrees by the health‐care team. Areas that focused on more practical aspects of nutritional support seemed to have a unified approach, whereas areas on nutritional assessment and routine nutrition orders demonstrated both a lack of knowledge and unclear role responsibilities. The results reveal how different professional groups in health care perceive the implementation of a clinical guideline. Some areas need further clarification, there needs to be continuing development of nutritional guidelines, and education of staff is needed in order to enhance the nutritional care of patients.
Development of indicators to assess the quality of nursing care in Thailand
Authors: Kunaviktikul Wipada , Anders Robert L. , Chontawan Ratanawadee , Nuntasupawat Ramual , Srisuphan Wichit , Pumarporn Orapun , Hanuchareonkul Somchit , Hirunnuj Sommai
Published: November 2005

Abstract  In Thailand, quality of nursing care has been defined. The next step is to determine how to measure quality. Given the limited resources to provide health care and the demands to justify the use of registered nurses, there is an urgent need to demonstrate how professional nursing care makes a difference to outcomes of care. The purpose of this study was to further refine nursing indicators of quality by developing operational definitions, validating them and then determining their applicability in a variety of clinical settings. The process included three phases. The first phase used focus groups to identify and define indicators of quality nursing care and the second phase included testing these indicators in four settings to determine if data could be collected. Manuals were developed that defined the quality indicators and the scoring system to be used in the assessment. In the third phase, the findings were presented to a group of experts and minor changes were made to the indicators. Then, the indicators were categorized into three groups: structure, process, and outcome. The validated outcomes and manual for their assessment and monitoring were sent to all hospitals in Thailand. The study relied upon the findings from the four different hospitals. If the indicators were implemented in a wider variety of settings, other differences might have emerged. There is also a possibility that ‘best practice’ might not be reflective of the scientific basis of nursing practice. The findings generally support the initial work carried out in the USA. There is a need to continue to study how these indicators can improve patient outcomes.
Dynamic association of the Ca2+ channel α1A subunit and SNAP‐25 in round or neurite‐emitting chromaffin cells
Authors: Andrés‐Mateos Eva , Renart Jaime , Cruces Jesús , Solís‐Garrido Luisa M. , Serantes Rocío , de Lucas‐Cerrillo Ana M. , Aldea Marcos , García Antonio G. , Montiel Carmen
Published: November 2005

Although the specific interaction between synaptic protein SNAP‐25 and the α1A subunit of the Cav2.1 channels, which conduct P/Q‐type Ca2+ currents, has been confirmed in in vitro‐translated proteins and brain membrane studies, the question of how native proteins can establish this association in situ in developing neurons remains to be elucidated. Here we report data regarding this interaction in bovine chromaffin cells natively expressing both proteins. The two carboxyl‐terminal splice variants of the α1A subunit identified in these cells share a synaptic protein interaction (‘synprint’) site within the II/III loop segment and are immunodetected by a specific antibody against bovine α1A protein. Moreover, both α1A isoforms form part of the P/Q‐channels–SNARE complexes in situ because they are coimmunoprecipitated from solubilized chromaffin cell membranes by a monoclonal SNAP‐25 antibody. The distribution of α1A and SNAP‐25 was studied in round or transdifferentiated chromaffin cells using confocal microscopy and specific antibodies: the two proteins are colocalized at the cell body membrane in both natural cell types. However, during the first stages of the cell transdifferentiation process, SNAP‐25 migrates alone out to the developing growth cone and what will become the nerve endings and varicosities of the mature neurites; α1A follows and colocalizes to SNAP‐25 in the now mature processes. These observations lead us to propose that the association between SNAP‐25 and α1A during neuritogenesis might promote not only the efficient coupling of the exocytotic machinery but also the correct insertion of P/Q‐type channels at specialized active zones in presynaptic neuronal terminals.
Activity of individual reticulospinal neurons during different forms of locomotion in the lamprey
Authors: Zelenin Pavel V.
Published: November 2005

Lamprey (a lower vertebrate) can employ different modes of locomotion, i.e. swimming in open water and crawling in tight places. Swimming is due to the periodic waves of lateral undulations with reciprocal activity of right and left muscles. In contrast, crawling (forward and backward) is based on single waves with coactivation of muscles on two sides. Basic mechanisms of swimming and, most likely, crawling reside in the spinal cord, and are activated by supraspinal commands. The main source of these commands is the reticulospinal (RS) system. The goal of the present experiments was to characterize the activity of individual RS neurons during swimming and during crawling in a U‐shaped tunnel. The activity was recorded by means of chronically implanted electrodes in freely behaving animals. All recorded RS neurons were active during swimming but silent in quiescent animals. Many of them (61%) showed phasic modulation of their firing rate approximately in phase with the activity of ipsilateral rostral muscles. The majority of the neurons (80%) were also active during crawling. Many of them either increased or decreased their activity during crawling as compared to the background activity. These changes were better correlated with the direction of progression (forward or backward) than with the direction of turning in the tunnel (right or left). No correlation of the activity of RS neurons during locomotion and their sensory inputs was found. The results of this study suggest that different modes of locomotion in lampreys can be caused by considerably overlapping groups of RS neurons.
Melatonin inhibits hippocampal long‐term potentiation
Authors: Wang Louisa M. , Suthana Nanthia A. , Chaudhury Dipesh , Weaver David R. , Colwell Christopher S.
Published: November 2005

The goal of this study is to investigate the effect of the hormone melatonin on long‐term potentiation and excitability measured by stimulating the Schaffer collaterals and recording the field excitatory postsynaptic potential from the CA1 dendritic layer in hippocampal brain slices from mice. Application of melatonin produced a concentration‐dependent inhibition of the induction of long‐term potentiation, with a concentration of 100 nm producing an ≈ 50% inhibition of long‐term potentiation magnitude. Long‐duration melatonin treatments of 6 h were also effective at reducing the magnitude of long‐term potentiation. Melatonin (100 nm) did not alter baseline evoked responses or paired‐pulse facilitation recorded at this synapse. The inhibitory actions of melatonin were prevented by application of the melatonin (MT) receptor antagonist luzindole as well as the MT2 receptor subtype antagonist 4‐phenyl‐2‐propionamidotetraline. These inhibitory actions of melatonin were lost in mice deficient in MT2 receptors but not those deficient in MT1 receptors. In addition, application of the protein kinase A inhibitor H‐89 both mimicked the effects of melatonin and precluded further inhibition by melatonin. Finally, the application an activator of adenylyl cyclase, forskolin, overcame the inhibitory effects of melatonin on LTP without affecting the induction of long‐term potentiation on its own. These results suggest that hippocampal synaptic plasticity may be constrained by melatonin through a mechanism involving MT2‐receptor‐mediated regulation of the adenylyl cyclase–protein kinase A pathway.
Movement‐related changes in oscillatory activity in the human subthalamic nucleus: ipsilateral vs. contralateral movements
Authors: Alegre M. , Alonso‐Frech F. , Rodríguez‐Oroz M. C. , Guridi J. , Zamarbide I. , Valencia M. , Manrique M. , Obeso J. A. , Artieda J.
Published: November 2005

A voluntary movement is accompanied by a series of changes in neuronal oscillatory activity in the subthalamic nucleus (STN). These changes can be recorded through electrodes implanted for deep brain stimulation to treat Parkinson's disease in the time interval between the surgery and the internalization of the connections to the batteries. Both baseline activity and movement‐related changes are different in the ‘on’ and ‘off’ medication motor states. In the ‘off’ state a low frequency activity in the alpha–beta range (8–25 Hz) that dominates the spectrum is interrupted during the movement, while in the ‘on’ state baseline frequencies are higher and a peri‐movement gamma increase (70–80 Hz) is usually observed. Similar changes have been described with electrocorticographic recordings over the primary motor cortex but the gamma increase was only present during contralateral movements. We compared ipsi‐ and contralateral movement‐related changes in STN activity, using a time–frequency analysis of the recordings obtained simultaneously in both STN and the scalp (electroencephalography) during right and left hand movements. The movement‐related changes observed in the STN in the ‘on’ and the ‘off’ states were similar to those described previously in terms of predominant frequency bands, but we found bilateral changes in the STN during movements of either hand. A contralateral earlier start of the beta STN changes was mostly observed when the moving hand corresponded to the less‐affected side, irrespective of hand dominance. These results suggest that movement‐related activity in the STN has, by and large, a bilateral representation and probably reflects cortical input.