bluuebunny/crossref_metadata_2025 huggingface dataset:
Self‐compartmentalized bacterial proteases and pathogenesis
Authors: Butler Susan M. , Festa Richard A. , Pearce Michael J. , Darwin K. Heran
Published: March 2006

Protein degradation is required for homeostasis of all living organisms. Self‐compartmentalized ATP‐dependent proteases are required for virulence of several pathogenic bacteria. Among the proteases implicated are ClpP and Lon, as well as the more recently identified bacterial proteasome. It is generally assumed that when a pathogen invades a host, microbial proteins become irreversibly damaged and need to be degraded. However, recent data suggest that proteolysis is also essential for virulence gene regulation. In this review, we will discuss what is known about the relationship between ATP‐dependent proteolysis and pathogenesis. In addition, we will propose other potential roles these chambered proteases may have in bacterial virulence. Importantly, these proteases show promise as targets for antimicrobial therapy.
The guanosine tetraphosphate (ppGpp) alarmone, DksA and promoter affinity for RNA polymerase in regulation of σ54‐dependent transcription
Authors: Bernardo Lisandro M. D. , Johansson Linda U. M. , Solera Dafne , Skärfstad Eleonore , Shingler Victoria
Published: March 2006

The RNA polymerase‐binding protein DksA is a cofactor required for guanosine tetraphosphate (ppGpp)‐responsive control of transcription from σ70 promoters. Here we present evidence: (i) that both DksA and ppGpp are required for in vivoσ54 transcription even though they do not have any major direct effects on σ54 transcription in reconstituted in vitro transcription and σ‐factor competition assays, (ii) that previously defined mutations rendering the housekeeping σ70 less effective at competing with σ54 for limiting amounts of core RNA polymerase similarly suppress the requirement for DksA and ppGpp in vivo and (iii) that the extent to which ppGpp and DksA affect transcription from σ54 promoters in vivo reflects the innate affinity of the promoters for σ54‐RNA polymerase holoenzyme in vitro. Based on these findings, we propose a passive model for ppGpp/DksA regulation of σ54‐dependent transcription that depends on the potent negative effects of these regulatory molecules on transcription from powerful stringently regulated σ70 promoters.
Lipid trafficking to the outer membrane of Gram‐negative bacteria
Authors: Doerrler William T.
Published: March 2006

The envelope of Gram‐negative bacteria is composed of two distinct lipid membranes: an inner membrane and outer membrane. The outer membrane is an asymmetric bilayer with an inner leaflet of phospholipids and an outer leaflet of lipopolysaccharide. Most of the steps of lipid synthesis occur within the cytoplasmic compartment of the cell. Lipids must then be transported across the inner membrane and delivered to the outer membrane. These topological features combined with the ability to apply the tools of biochemistry and genetics make the Gram‐negative envelope a fascinating model for the study of lipid trafficking. In addition, as lipopolysaccharide is essential for growth of most strains and is a potent inducer of the mammalian innate immune response via activation of Toll‐like receptors, Gram‐negative lipid transport is also a promising target for the development of novel antibacterial and anti‐inflammatory compounds. This review focuses on recent developments in our understanding of lipid transport across the inner membrane and to the outer membrane of Gram‐negative bacteria.
Identification of Bacillus subtilis	σW‐dependent genes that provide intrinsic resistance to antimicrobial compounds produced by Bacilli
Authors: Butcher Bronwyn G. , Helmann John D.
Published: March 2006

Bacillus subtilis produces many antibiotics of varying structures and specificity. Here we identify a prominent role for σW, an extracytoplasmic function (ECF) sigma factor, in providing intrinsic resistance to antimicrobial compounds produced by other Bacilli. By using a panel of B. subtilis mutants disrupted for each of the 30 known σW‐dependent operons we identified resistance genes for at least three different antimicrobial compounds. The ydbST and fosB genes contribute to resistance to antimicrobial compound(s) produced by B. amyloliquefaciens FZB42, the yqeZyqfAB operon provides resistance to the SPβ prophage‐encoded bacteriocin sublancin, and the yknWXYZ operon and yfhL provide resistance to the antimicrobial peptide SdpC. YfhL encodes a paralogue of SdpI, a membrane protein that provides immunity to SdpC. In competition experiments, we identify σW as a key factor in allowing B. subtilis to resist antibiotic killing and encroachment by competing strains. Together with the previous observation that σW provides inducible resistance against the Streptomyces antibiotic fosfomycin, these studies support the notion that σW controls an antibiosis regulon important in the microbial ecology of soil bacteria.
Oligonucleotide cleavage and rejoining by topoisomerase III from the hyperthermophilic archaeon Sulfolobus solfataricus: temperature dependence and strand annealing‐promoted DNA religation
Authors: Chen Liang , Huang Li
Published: March 2006

Topoisomerase III from the hyperthermophilic archaeon Sulfolobus solfataricus (Sso topo III) is optimally active in DNA relaxation at 75°C. We report here that Sso topo III‐catalysed DNA cleavage and religation differed significantly in temperature dependence: the enzyme was most active in cleaving ssDNA containing a cleavage site at 25–50°C, but was efficient in rejoining the cleaved DNA strand only at higher temperatures (e.g. ≥ 45°C). The failure of Sso topo III to rejoin the cleaved DNA strand efficiently appeared to be responsible for the inability of the enzyme to relax negatively supercoiled DNA at low temperature (e.g. 25°C). Intriguingly, Sso topo III facilitated DNA annealing although it showed higher affinity for ssDNA than for dsDNA. Religation of the DNA strand cleaved by Sso topo III was drastically enhanced when the DNA was allowed to anneal to a complementary non‐cleaved oligonucleotide, presumably as a result of destabilization of the interaction between the enzyme and the cleaved strand through the formation of duplex DNA. A region in the non‐cleaved strand corresponding to a sequence containing six bases on the 5′ side and two bases on the 3′ side of the cleavage site in the cleaved strand was crucial to the annealing‐promoted religation. However, the annealing‐promoted religation was relatively insensitive to mismatches in this region and the region conserved for oligonucleotide cleavage, except for that at the 5′ end of the broken strand. These results suggest that Sso topo III is well suited for a role in DNA rewinding, whether it leads to homoduplex or heteroduplex formation.
HLA and enteric antineuronal antibodies in patients with achalasia
Authors: Latiano A. , De giorgio R. , Volta U. , Palmieri O. , Zagaria C. , Stanghellini V. , Barbara G. , Mangia A. , Andriulli A. , Corinaldesi R. , Annese V.
Published: March 2006

Abstract  The aetiopathogenesis of primary achalasia is largely unknown, although an immunogenetic predisposition is suspected. To establish whether a correlation exists among possible aetiological factors, including class II human leucocyte antigen (HLA) alleles and autoantibodies to enteric neurones, and clinical features of patients with achalasia, a total of 60 patients and 200 healthy subjects were typed by high‐resolution HLA‐DQ and HLA‐DR alleles. Circulating antineuronal antibodies were investigated by using indirect immunofluorescence on enteric neurones of rat ileum and colon and immunoblotting assay in a subset of achalasic patients and in all controls. The DQB1*0502 and DQB1*0601 alleles were significantly increased in patients with achalasia compared with controls (P < 0.03, P < 0.001, respectively). Moreover a negative correlation with the DQB1*0201 allele was found (P = 0.016). As a whole, 14 of 60 (23.3%) achalasia patients were carriers of HLA risk alleles, and 10 of 41 (24.4%) presented antineuronal antibodies. No significant correlation among HLA risk alleles, antineuronal antibodies and clinical features was found. In achalasia, no correlation exists among HLA alleles, antineuronal antibodies and clinical features. However, given the association between achalasia and HLA‐DQ1, further research is needed to clarify the role of HLA antigens and antineuronal antibodies in this disease.
Modification of intracellular levels of glutathione‐dependent formaldehyde dehydrogenase alters glutathione homeostasis and root development
Authors: ESPUNYA M. CARME , DÍAZ MAYKELIS , MORENO‐ROMERO JORDI , MARTÍNEZ M. CARMEN
Published: March 2006

Glutathione (GSH)‐dependent formaldehyde dehydrogenase (FALDH) is a highly conserved medium‐chain dehydrogenase reductase and the main enzyme that metabolizes intracellular formaldehyde in eukaryotes. It has been recently shown that it exhibits a strong S‐nitrosoglutathione (GSNO) reductase activity and could be a candidate to regulate NO‐signalling functions. However, there is a lack of knowledge about the tissue distribution of this enzyme in plants. Here, we have studied the localization and developmental expression of the enzyme using immunolocalization and histochemical activity assay methods. We conclude that FALDH is differentially expressed in the organs of Arabidopsis thaliana mature plants, with higher levels in roots and leaves from the first stages of development. Spatial distribution of FALDH in these two organs includes the main cell types [epidermis (Ep) and cortex (Cx) in roots, and mesophyll in leaves] and the vascular system. Arabidopsis thaliana mutants with modified levels of FALDH (both by over‐ and under‐expression of the FALDH‐encoding gene) show a significant reduction of root length, and this phenotype correlates with an overall decrease of intracellular GSH levels and alteration of spatial distribution of GSH in the root meristem. Transgenic roots are partially insensitive to exogenous GSH, suggesting an inability to detect reduction–oxidation (redox) changes of the GSH pool and/or maintain GSH homeostasis.
The Reduction of Serum B‐Lymphocyte Activating Factor levels following Quinacrine Add‐On Therapy in Systemic Lupus Erythematosus
Authors: Toubi E. , Kessel A. , Rosner I. , Rozenbaum M. , Paran D. , Shoenfeld Y.
Published: March 2006

B‐Lymphocyte‐activating factor (BAFF/BLyS) is a survival factor for B cells, belonging to the tumor necrosis ligand super family. Serum BAFF levels have been found to be elevated in patients with systemic lupus erythematosus (SLE). Neutralization of BAFF activity was suggested as an additional therapeutic approach in SLE. To determine the effect of add‐on Quinacrine (Qn) treatment on serum BAFF levels and the effect of this treatment on SLE disease activity index (SLEDAI), antidsDNA and anticardiolipin (aCL) antibody levels, we treated 29 stable SLE patients, who were maintained on prednisolone and hydroxychloroquine and in some on azathioprine (AZT), with additional Qn (100 mg/d) with an aim to further reduce disease activity. SLEDAI, antidsDNA, aCL antibodies and serum BAFF levels were assessed before and 3 months after the addition of Qn. Three months following Qn initiation, a reduction in SLEDAI was noticed in 19/29 patients (mean 8.8 ± 2.3 to 3.3 ± 1.5,P = 0.009), followed by reduction or discontinuation of prednisolone in all patients and the discontinuation of AZT in five patients. Serum BAFF levels were significantly reduced in 8/12 patients (mean 6.3 ± 0.5 to 3.0 ± 0.56 ng/mlP = 0.0001). This reduction was found in correlation with a decrease in aCL titres. However, the decrease in SLEDAI scores and antidsDNA antibody titres was unrelated to the decrease in serum BAFF or aCL levels. We conclude that the addition of Qn to previous therapeutic regimens in active SLE is beneficial and seems to reduce SLEDAI scores, serum BAFF and aCL levels and therefore should be considered in many of our SLE patients before aggressive treatments are given.
Transient Appearance of Hepatic Natural Killer Cells with High Cytotoxicity and Unique Phenotype in Very Young Mice
Authors: Bai X. , Wang S. , Tomiyama‐Miyaji C. , Shen J. , Taniguchi T. , Izumi N. , Li C. , Bakir H. Y. , Nagura T. , Takahashi S. , Kawamura T. , Iiai T. , Okamoto H. , Hatakeyama K. , Abo T.
Published: March 2006

There were few natural killer (NK) cells in the liver in very young mice at the age of 1–2 weeks. This was because the cell yield from the liver of young mice was low. The percentage of NK cells in the liver of young mice, however, was almost comparable with that in the liver of adult mice. Lymphocytes were isolated from the liver and spleen of C57BL/6 (B6) mice, and NK cytotoxicity and phenotype were herein examined in this study. NK cytotoxicity was extremely high in the liver of very young mice. This phenomenon was seen in the liver of various normal mouse strains. In contrast, the appearance of high cytotoxicity was not seen in NK cells of the spleen, irrespective of mouse strains. The quality of NK cells in the liver of young mice was different from that in adult mice. NK cells in the liver of young mice were mainly CD69+Mac‐1– Fas ligand+, whereas those in the liver of adult mice were CD69–Mac‐1+ Fas ligand–. These results revealed that the quality of hepatic NK cells changes in the process of ageing. Namely, liver NK cells in very young mice temporarily show the highest NK cytotoxicity and a unique activated phenotype. Physiological meaning of the present phenomenon was discussed.
High‐Altitude Climate Therapy Reduces Local Airway Inflammation and Modulates Lymphocyte Activation
Authors: Karagiannidis C. , Hense G. , Rueckert B. , Mantel P. Y. , Ichters B. , Blaser K. , Menz G. , Schmidt‐Weber C. B.
Published: March 2006

High‐altitude climate therapy is a well‐established therapeutic option, which improves clinical symptoms in asthma. However, little is known about the underlying immunological mechanisms. The study investigates the influence of high‐altitude climate therapy on airway inflammation and cellular components of specific and unspecific immune response. Exhaled NO significantly decreased within 3 weeks of therapy in patients with allergic and intrinsic, moderate and severe asthma. Interleukin‐10 (IL‐10)‐secreting peripheral blood mononuclear cells (PBMC) increased within 3 weeks of therapy in six of 11 patients, whereas transforming growth factor‐β1‐secreting PBMC remained stable. Furthermore, monocyte activation, assessed by CD80 expression significantly decreased during therapy. The frequency of CRTH2‐expressing T cells decreased, while regulatory T cells (Treg) remained stable. FOXP3 and GATA‐3 mRNA expression in CD4+ T cells did not change, while interferon‐γ and IL‐13 mRNA expression decreased in eight of 10 patients. The current data demonstrate that high‐altitude climate therapy reduces local airway inflammation. Furthermore, monocytes switch towards a tolerogenic phenotype under high‐altitude climate therapy. The Treg/Th2 ratio increases; however, because of the absence of antigens/allergens, no de novo differentiation of Th2 nor Treg cells is observed. The high‐altitude climate therapy therefore may form the immunological basis for the endogenous control of allergen‐driven diseases.
Human Cell Lines Used in a Micro Neutralization Test for Measuring Influenza‐Neutralizing Antibodies
Authors: Mittelholzer C. M. , Brokstad K. A. , Pauksens K. , Jonsson R. , Brytting M. , Linde A.
Published: March 2006

An in situ neutralization test (NT) including ELISA for the measurement of influenza antigen was developed and evaluated. Two human cell lines, fibroblasts (HS27) cells and salivary gland epithelial duct (HSG) cells, were compared with Madin‐Darby Canine Kidney (MDCK) cells. The viral production in the human cell lines was lower than that for MDCK cells, which influenced the results of the assay in the HSG and HS27 cells. However, when lowering the infectious dose, the NT using HS27 cells gave a sensitive and stable assay with low background in the ELISA. The NT titres were very low when using HSG cells compared to MDCK cells. The HS27 NT was used to analyze the humoral response after an influenza A infection in patients from a placebo‐controlled zanamivir study. We found no differences in NT titres between patients treated with zanamivir or placebo. The MDCK and HS27 NT gave higher titres and more pronounced titre differences than the gold standard haemagglutinin inhibition (HAI) assay. Compared to the HAI assay, the sensitive NT using HS27 cells also revealed heterologous NT‐titre rises after influenza infection in the patients.
Significance of Alterations in PBMC Immunophenotype of Children with Chronic Viral Hepatitis C – the Role of Dendritic Cells
Authors: Mozer‐Lisewska I. , Dworacki G. , Kaczmarek E. , Slużewski W. , Kaczmarek M. , Woźniak A. , Żeromski J.
Published: March 2006

There are differences in the clinical course of chronic viral hepatitis C between adults and children, but it is generally accepted that the disease has cell‐mediated immune background. The aim of this study was to evaluate PBMC subsets in children with chronic hepatitis C before treatment in order to find some predictive factors, useful for patients management. Several PBMC subsets, in particular lymphoid and dendritic cell (DC) ones, were tested by flow cytometry in HCV+ paediatric patients (n = 46) and in control children matched in terms of age and sex (n = 20). Data were subjected to extensive statistics. It was found that cells with cytotoxic potential such as CD8+CD28– T cells, NK and NKT cells as well as lineage–HLA‐DR+ DC were increased in per cent values, while CD4+ T cells and CD4:CD8 ratio were decreased in hepatitis C group. In HCV+ patients, CD4+ T cells were inversely correlated with alanine aminotransferase (ALT) levels and with viraemia. DC subset of myeloid origin (CD11c+) assessed both in per cent values and as mean fluorescence intensity (MFI) of HLA‐DR expression was shown to be downregulated in hepatitis patients, in spite of increased numbers. To conclude, PBMC subsets, and in particular DC, are affected by HCV chronic infection in children, reflected by the correlation with clinical parameters, such as ALT and viraemia.
Interplay between erosion and tectonics in the Western Alps
Authors: Malusà Marco G. , Vezzoli Giovanni
Published: March 2006

Bedrock fission‐track analysis, high‐resolution petrography and heavy mineral analyses of sediments are used to investigate the relationships between erosion and tectonics in the Western Alps. Along the Aosta Valley cross‐section, exhumation rates based on fission‐track data are higher in the fault‐bounded western block than in the eastern block (0.4–1.5 vs. 0.1–0.3 mm yr−1). Erosion rates based on the analysis of bed‐load in the Dora Baltea drainage display the same pattern and have similar magnitudes in the relative sub‐basins (0.4–0.7 vs. 0.04–0.08 mm yr−1). Results highlight that climate, relief and lithology are not the controlling factors of erosion in the Western Alps. The main driving force behind erosion is instead tectonics that causes the differential upward motion of crustal blocks.
Delamerian‐aged metamorphism in the southern Curnamona Province, Australia: implications for the evolution of the Mesoproterozoic Olarian Orogeny
Authors: Rutherford Lachlan , Hand Martin , Mawby Joanna
Published: March 2006

Monazite electron microprobe U–Th–Pb and garnet Sm–Nd isotopic data from metapelitic assemblages in the Willyama Supergroup in the southern Curnamona Province, south‐central Australia, indicate that the terrain underwent regional greenschist to amphibolite‐grade metamorphism during the c. 500 Ma Delamerian Orogeny. The Delamerian‐aged mineral assemblages include prograde garnet–staurolite and kyanite‐bearing associations that overprint andalusite‐ and sillimanite‐bearing assemblages that are interpreted to have developed during the c. 1600 Ma Olarian Orogeny. Importantly, the development of secondary kyanite‐bearing assemblages in the southern Curnamona Province has been used previously to suggest that the Olarian Orogeny followed an anticlockwise P–T evolution. If such assemblages are the product of c. 500 Ma metamorphism, then the anticlockwise P–T path is an apparent path, due to the overprint of a distinct metamorphic cycle c. 1100 Ma later. Making such distinctions is therefore extremely important when using the textural and metamorphic evolution of polycyclic terrains to model the thermal behaviour of the crust during orogeny. This study highlights the utility of in situ geochronology, linking age data to petrologically important phases and assemblages.
50 Myr recovery from the largest negative δ13C excursion in the Ediacaran ocean
Authors: Le Guerroué Erwan , Allen Philip A. , Cozzi Andrea , Etienne James L. , Fanning Mark
Published: March 2006

Sedimentary rocks deposited during the Ediacaran period (∼630–542 Ma) contain carbonates whose carbon isotopic ratios show a marked negative excursion consisting of a precipitous drop from +5‰ to −12‰, followed by a sub‐linear recovery to positive δ13C values. Isotopic ages (U/Pb) and thermal subsidence modelling are combined to constrain the excursion in time and indicate an onset at ∼600 Ma, and duration of recovery of approximately 50 Myr. The excursion is widely recognized in Oman and has potential correlatives in Ediacaran strata elsewhere, and may thus represent a characteristic feature of the Ediacaran period. The amplitude of this carbon isotope excursion far exceeds those of other Neoproterozoic anomalies. The isotopic trend of negative excursion and long‐term recovery spanned at least one short‐lived glacial episode (at 580 Ma), but appears unrelated to glaciation, which indicates that negative anomalies in the Neoproterozoic marine carbon isotope record are not directly or uniquely linked to ice ages.
Simultaneous flux and current measurement from single plant protoplasts reveals a strong link between K+ fluxes and current, but no link between Ca2+ fluxes and current
Authors: Gilliham Matthew , Sullivan Wendy , Tester Mark , Tyerman Stephen D.
Published: March 2006

We present a thorough calibration and verification of a combined non‐invasive self‐referencing microelectrode‐based ion‐flux measurement and whole‐cell patch clamp system as a novel and powerful tool for the study of ion transport. The system is shown to be capable of revealing the movement of multiple ions across the plasma membrane of a single protoplast at multiple voltages and in complex physiologically relevant solutions. Wheat root protoplasts are patch clamped in the whole‐cell configuration and current–voltage relations obtained whilst monitoring net K+ and Ca2+ flux adjacent to the membrane with ion‐selective electrodes. At each voltage, net ion flux (nmol m−2 sec−1) is converted to an equivalent current density (mA m−2) taking into account geometry and electrode efficiency, and compared with the net current density measured with the patch clamp system. Using this technique, it is demonstrated that the K+‐permeable outwardly rectifying conductance (KORC) is responsible for net outward K+ movement across the plasma membrane [1:1 flux‐to‐current ratio (1.21 ± 0.14 SEM, n = 15)]. Variation in the K+ flux‐to‐current ratio among single protoplasts suggests a heterogeneous distribution of KORC channels on the membrane surface. As a demonstration of the power of the technique we show that despite a significant Ca2+ permeability being associated with KORC (analysis of tail current reversal potentials), there is no correlation between Ca2+ flux and KORC activity. A very significant observation is that large Ca2+ fluxes are electrically silent and probably tightly coupled to compensatory charge movements. This analysis demonstrates that it is mandatory to measure flux and currents simultaneously to investigate properly Ca2+ transport mechanisms and selectivity of ion channels in general.
Sequencing and characterization of telomere and subtelomere regions on rice chromosomes 1S, 2S, 2L, 6L, 7S, 7L and 8S
Authors: Mizuno Hiroshi , Wu Jianzhong , Kanamori Hiroyuki , Fujisawa Masaki , Namiki Nobukazu , Saji Shoko , Katagiri Satoshi , Katayose Yuichi , Sasaki Takuji , Matsumoto Takashi
Published: March 2006

Telomeres, which are important for chromosome maintenance, are composed of long, repetitive DNA sequences associated with a variety of telomere‐binding proteins. We characterized the organization and structure of rice telomeres and adjacent subtelomere regions on the basis of cytogenetic and sequence analyses. The length of the rice telomeres ranged from 5.1 to 10.8 kb, as revealed by both fibre‐fluorescent in situ hybridization and terminal restriction‐fragment assay. Physical maps of the chromosomal ends were constructed from a fosmid library. This facilitated sequencing of the telomere regions of chromosomes 1S, 2S, 2L, 6L, 7S, 7L and 8S. The resulting sequences contained conserved TTTAGGG telomere repeats, which indicates that the physical maps partly covered the telomere regions of the respective chromosome arms. These repeats were organized in the order of 5′‐TTTAGGG‐3′ from the chromosome‐specific region, except in chromosome 7S, in which seven inverted copies also existed in tandem array. Analysis of the telomere‐flanking regions revealed the occurrence of deletions, insertions, or chromosome‐specific substitutions of single nucleotides within the repeat sequences at the junction between the telomere and subtelomere. The sequences of the 500‐kb regions of the seven chromosome ends were analysed in detail. A total of 598 genes were predicted in the telomeric regions. In addition, repetitive sequences derived from various kinds of retrotransposon were identified. No significant evidence for segmental duplication could be detected within or among the subtelomere regions. These results indicate that the rice chromosome ends are heterogeneous in both sequence and characterization.
Genome‐wide transcriptional analysis of the Arabidopsis thaliana interaction with the plant pathogen Pseudomonas syringae pv. tomato DC3000 and the human pathogen Escherichia coli O157:H7
Authors: Thilmony Roger , Underwood William , He Sheng Yang
Published: March 2006

Pseudomonas syringae pv. tomato DC3000 (Pst) is a virulent pathogen that causes disease on tomato and Arabidopsis. The type III secretion system (TTSS) plays a key role in pathogenesis by translocating virulence effectors from the bacteria into the plant host cell, while the phytotoxin coronatine (COR) contributes to virulence and disease symptom development. Recent studies suggest that both the TTSS and COR are involved in the suppression of host basal defenses. However, little is known about the interplay between the host gene expression changes associated with basal defenses and the virulence activities of the TTSS and COR during infection. In this study, we used the Affymetrix full genome chip to determine the Arabidopsis transcriptome associated with basal defense to Pst DC3000 hrp mutants and the human pathogenic bacterium Escherichia coli O157:H7. We then used Pst DC3000 virulence mutants to characterize Arabidopsis transcriptional responses to the action of hrp‐regulated virulence factors (e.g. TTSS and COR) during bacterial infection. Additionally, we used bacterial fliC mutants to assess the role of the pathogen‐associated molecular pattern flagellin in induction of basal defense‐associated transcriptional responses. In total, our global gene expression analysis identified 2800 Arabidopsis genes that are reproducibly regulated in response to bacterial pathogen inoculation. Regulation of these genes provides a molecular signature for Arabidopsis basal defense to plant and human pathogenic bacteria, and illustrates both common and distinct global virulence effects of the TTSS, COR, and possibly other hrp‐regulated virulence factors during Pst DC3000 infection.
New developmental insights from high‐throughput biological analysis in Caenorhabditis elegans
Authors: Labbé J‐C , Roy R
Published: March 2006

The use of Caenorhabditis elegans as a model system for understanding animal development and human disease has long been recognized as an efficient tool of discovery. Recent developments, particularly in our understanding of RNA‐mediated interference and its ability to modify gene activity, have facilitated the use of C. elegans in determining gene function via high‐throughput analysis. These new strategies have provided a framework that allows investigators to analyse gene function globally at the genomic level and will likely become a prototypic model for biological analysis in the post‐genome era.
Gene frequencies of human platelet antigens in the Macedonian population
Authors: Pavkovic M. , Petlichkovski A. , Strezova A. , Arsov T. , Trajkov D. , Spiroski M.
Published: March 2006

Human platelet antigen (HPA) systems consist of more than 12 bi‐allelic antigen polymorphisms. Due to these polymorphisms, platelet‐membrane glycoproteins can be recognized as alloantigens or autoantigens and can cause conditions such as fetomaternal alloimmune thrombocytopenia, post‐transfusion refractoriness to platelets, and post‐transfusion throbocytopenic purpura. The purpose of this study was to investigate the distribution of HPA‐1, ‐2, ‐3, and ‐5 in Macedonian population by using the polymerase chain reaction and restriction fragment length polymorphism. The allele frequencies were 0.865 for HPA‐1a, 0.135 for HPA‐1b, 0.852 for HPA‐2a, 0.148 for HPA‐2b, 0.578 for HPA‐3a, 0.422 for HPA‐3b, 0.909 for HPA‐5a, and 0.091 for HPA‐5b. Results of our study were not significantly different from those reported in the other European studies. Our population displayed the highest frequency for HPA‐2b allele (0.148) reported among European population.
Optimization of the MhcOvar‐DRB1 gene typing
Authors: Arrieta‐Aguirre I. , García‐Etxebarria K. , Jugo B. M.
Published: March 2006

The variability of the sheep major histocompatibility complex (MHC) class II DRB1 locus has been analyzed in this work. Exon 2 of Ovar‐DRB1 was amplified by polymerase chain reaction (PCR) with the primers designed by Amills et al. for goats. In a total of 187 sheep of Latxa breed, we identified by PCR‐single‐strand conformation polymorphism (SSCP) 19 alleles, eight of them previously unpublished. Moreover, the observed heterozygosity reached 91–95%. A new allelic type named DRB*14 was defined, which brings to light once more the discussion as to the ancestor of the domestic sheep. Although all the defined alleles can be discriminated by PCR‐SSCP, sequence‐based typing is proposed as the technique of choice for sheep DRB1 gene typing.
HLA‐DRB1, ‐DQB1 alleles in head and neck carcinoma patients
Authors: Koskinen W. J. , Partanen J. , Vaheri A. , Aaltonen L.‐M.
Published: March 2006

Certain HLA class II alleles have been reported to play a role in development or prevention of cervical carcinoma, an epithelial malignancy linked to human papillomavirus (HPV). In head and neck carcinomas, of which a subset is also HPV associated, the impact of HLA genes remains unknown. HLA‐DRB1, ‐DQB1 alleles were determined in a comprehensive series of 162 head and neck carcinoma patients, for which 83 consecutive cadaveric organ donors of Finnish origin served as controls. DRB1*03 was associated with node‐negative disease and DRB1*08 and 13 with small tumors; DRB1*04 was protective against disease relapse. Most alleles of borderline significance in this study act similarly in cervical carcinomas.
Th1/Th2 cytokine gene polymorphisms in patients with idiopathic pulmonary fibrosis
Authors: Vasakova M. , Striz I. , Slavcev A. , Jandova S. , Kolesar L. , Sulc J.
Published: March 2006

We investigated 30 patients with idiopathic pulmonary fibrosis (IPF) and 103 healthy volunteers for the cytokines polymorphisms of the IL‐1α, IL‐1β, IL‐1R, IL‐1RA, IL‐2, IL‐4, IL‐6, IL‐10, IL‐12, tumor necrosis factor‐α, interferon‐γ, transforming growth factor‐β, IL‐1β, IL‐2, IL‐4, and IL‐4RA genes. The strongest correlation of a genotype with the disease was found for gene polymorphisms at the promotor region of IL‐4, where the CT genotypes at the positions (−590) and (−33) were more frequent in the IPF group (P < 0.0001,Pcorr < 0.0022;vs P < 0.0001,Pcorr < 0.0022). Our results support the idea of the pathogenic role of cytokine gene polymorphisms in the etiology and pathogenesis of IPF, with emphasize on the IL‐4 promotor gene polymorphisms.
Investigation of killer cell immunoglobulin‐like receptor KIR2DL4 diversity by sequence‐based typing in Chinese population
Authors: Zhu F.‐M. , Jiang K. , Lv Q.‐F. , He J. , Yan L.‐X.
Published: March 2006

Human killer cell immunoglobulin‐like receptors (KIRs) play an important role in controlling natural killer (NK) cell function. Here, polymerase chain reaction sequence‐based typing (PCR‐SBT) procedures identifying alleles of the KIR2DL4 gene have been established. The method was designed around the specific amplification of exon 3 to exon 5 and exon 7 to exon 9 of the KIR2DL4 gene and produce discrimination of KIR2DL4 alleles. Genomic DNAs from 83 healthy unrelated Chinese Han individuals were typed for KIR2DL4 alleles by this method. Each sample was assigned to the putative KIR2DL4 allele combination according to the nucleotide polymorphism profiles of all KIR2DL4 alleles. Twenty‐one different genotypes and seven KIR2DL4 alleles were observed in the population, with KIR2DL4*00102 having the highest frequency, 0.5. Five individuals bear a recombinant allele KIR3DP*004 that associated with three putative KIR2DL4 alleles. Our data demonstrated that the established PCR‐SBT method for KIR2DL4 allele typing was reliable, and Chinese Han population is distinct in KIR2DL4 allele frequencies in comparison to some other populations.
Identification of a novel HLA‐B*44 variant (B*4441) in three unrelated Caucasian individuals
Authors: Hoelsch K. , Lenggeler I. , Knabe H. , Hartel K. , Klein H‐G. , Woelpl A.
Published: March 2006

Here, we report the identification of a new human leukocyte antigen (HLA)‐B*44 allele found almost simultaneous in three DNA samples which were part of routine bone marrow donor typing by order of the German registry ‘Aktion Knochenmarkspende Bayern’. The samples appeared noticeable in different polymerase chain reactions using sequence‐specific primers (PCR‐SSP) or sequence‐specific oligonucleotides (PCR‐SSO). Sequence‐based typing revealed a novel allele officially designated as B*4441*. This sequence differs from HLA‐B*44020101/4427 by two nucleotide positions at the beginning of exon 3: by position 353 (T to C) and by position 355 (A to C). These differences in sequence result in deviant amino acids at codon 94 (Ile94Thr) and codon 95 (Ile95Leu).
Eliciting repeat blood donations: tell early career donors why their blood type is special and more will give again
Authors: Chamla J. H. , Leland L. S. , Walsh K.
Published: March 2006

Background and Objectives  This study was designed to investigate whether sending a personalized and informative letter to early career donors would increase the number returning to donate again, as the literature suggests that it is at around the third donation that people become career donors.

Materials and Methods  Experimental participants were sent a recruitment letter that included information about their own blood type and the percentage of the general population with the same blood type who donated. Control participants received a recruitment letter with some general information. The aim of the study was to determine whether more participants in the experimental group, compared with the control group, would donate blood within a 4‐week period after receiving their personalized recruitment letter.

Results  Donors in the experimental group were 43% more likely to return to donate than those in the control group [χ2 (1) = 5·79, P < 0·016].

Conclusions  Sending out personalized, informative letters appears to be a potentially powerful donor‐retention tool.
Treatment of adult patients with sepsis‐induced coagulopathy and purpura fulminans using a plasma‐derived protein C concentrate (Ceprotin®)
Authors: Schellongowski Peter , Bauer Edith , Holzinger Ulrike , Staudinger Thomas , Frass Michael , Laczika Klaus , Locker Gottfried J. , Quehenberger Peter , Rabitsch Werner , Schenk Peter , Knöbl Paul
Published: March 2006

Background and Objectives  The aim of this study was to document the effects of supplementation with a plasma‐derived protein C concentrate in adult patients with infectious purpura fulminans.

Materials and Methods  We report the effect of the administration of a human protein C concentrate (Ceprotin®, Baxter, Vienna, Austria) in eight adult patients with purpura fulminans. Five patients received the concentrate as level‐adjusted continuous infusion (10 U/kg/h, target protein C activity 100%) and three patients received the concentrate as bolus infusions (100 U/kg every 6 h) in addition to standard sepsis therapy. Heparin, fresh‐frozen plasma, antithrombin‐ and fibrinogen concentrates, low‐dose rtPA, and platelet transfusions were given when appropriate.

Results  Six patients had overt disseminated intravascular coagulation: platelets, 19 g/l; fibrinogen, 60 mg/dl; antithrombin, 47%; prothrombin time, 32%; activated partial thromboplastin time (APTT), 88 s; d‐dimer, 66 µg/ml; protein C activity, 29% (medians). Five patients had septic shock, six renal failure and four respiratory failure. Patients received between 5000 and 77 000 U of protein C concentrate over 2·5 days (median); the protein C activity increased to 184% (median) and coagulopathy resolved within 3 days in seven of the eight patients. Six patients survived, one died early from fulminant sepsis, and one died after 14 days from candida sepsis.

Conclusions  Our data suggest that treatment with a plasma‐derived protein C zymogen concentrate might be a useful support in adult patients with purpura fulminans.
A Study of Dry Cow Therapy and Effects on SCC in 10 Irish Dairy Herds
Authors: Barrett D. J. , Clegg T. , Healy A. M. , Doherty M. L.
Published: March 2006

Somatic cell count (SCC) data for 480 cows in 10 Irish dairy herds from January 2001 until June 2002 were analysed. Herds were selected on the basis of a recent or ongoing history of clinical or subclinical mastitis. An individual cow SCC of 200 000 cells per ml was used as the threshold for elevation of SCC. The duration of elevated SCC prior to drying‐off and the magnitude of the elevation in SCC were found to have an impact on the response to dry cow therapy (DCT). A trend also emerged indicating that increasing parity had a negative influence on the response to DCT.
Efficacy of Oral Tadalafil, a New Long‐acting Phosphodiesterase‐5 Inhibitor, for the Short‐term Treatment of Pulmonary Arterial Hypertension in a Dog
Authors: Serres F. , Nicolle A. P. , Tissier R. , Gouni V. , Pouchelon J.‐L. , Chetboul V.
Published: March 2006

Systolic pulmonary arterial hypertension (PAH) was diagnosed in a 15‐year‐old intact male Yorkshire terrier presented for progressive dyspnoea and coughing. Several examinations were performed (thoracic radiographs, faecal analysis, heartworm antigen test, tracheal fluoroscopy, abdominal ultrasound, complete blood cell count, urine and serum biochemistry) but the PAH remained of unknown origin. Despite medical treatment (diuretics and angiotensin‐converting enzyme inhibitor), cardiovascular and respiratory signs dramatically worsened over a 1‐month period, with several daily syncope, cyanosis and tachypnoea at rest requiring permanent oxygen therapy. Oral tadalafil (Cialis®), a new long‐acting phosphodiesterase‐5 inhibitor, belonging to the same family as sildenafil (Viagra®), was added to the background therapy. The condition of the dog improved quickly (<24 h), and short‐term follow up (7 days) showed a decrease in systolic pulmonary arterial pressure up to 26 mmHg concomitant with the disappearance of all respiratory and cardiac signs of PAH (cyanosis, syncope and tachypnoea). This case is of interest because it concerns the first reported short‐term use of tadalafil in canine PAH. However, long‐term studies with a large number of diseased animals are now required before prescription by general practitioners could be recommended.
Comparison of Lipase Activity in Peritoneal Fluid of Dogs with Different Pathologies – A Complementary Diagnostic Tool in Acute Pancreatitis?
Authors: de Arespacochaga A. Guija , Hittmair K. M. , Schwendenwein I.
Published: March 2006

A clinical diagnosis of acute pancreatitis is often difficult to obtain. Histopathology remains the gold standard, whereas clinical signs, diagnostic imaging and laboratory testing, even in combination, may be insufficient. In a prospective study, lipase activity in ascitic fluid of various aetiologies was determined in 44 dogs in order to investigate its performance in cases of acute pancreatitis. Data of simultaneously determined blood lipase activities were available in 27 dogs. Lipase activity was measured by a colorimetric assay. A complete peritoneal fluid analysis was performed. Dogs were divided into four groups, according to their final diagnosis: acute pancreatitis (A), abdominal trauma (B), abdominal neoplasia (C) and others (hepatic or cardiac diseases) (D). Dogs with acute pancreatitis had a significantly higher peritoneal lipase activity than those in other groups (P ≤ 0.024), while no significant difference was found between the other groups (P ≥ 0.734). Blood lipase activity as well as protein content and total cell count of the ascitic fluid did not show any significant difference between groups. Data show that determination of lipase activity in dogs that develop ascites may be useful in complementing the diagnosis of acute pancreatitis.
Seroprevalence of Anaplasma phagocytophilum among Healthy Dogs and Horses in Israel
Authors: Levi O. , Waner T. , Baneth G. , Keysary A. , Bruchim Y. , Silverman J. , Harrus S.
Published: March 2006

The presence of reacting antibodies to Anaplasma phagocytophilum has previously been demonstrated in Israel, both in humans and the golden jackal (Canis aureus syriacus). This study was undertaken to determine the seroprevalence of A. phagocytophilum antibodies in two additional potential hosts, domestic dogs and horses in order to investigate the possibility of exposure to the organism in Israel. Of 195 dogs tested, 9% were seroreactive with A. phagocytophilum antigen and 30% were seroreactive to Ehrlichia canis. Twenty‐nine percent of the dogs seropositive for E. canis were also reactive to A. phagocytophilum. Two dogs had immunofluoresence antibody (IFA) antibody titres for A. phagocytophilum greater than E. canis. The equine serological survey (n = 300) revealed no seroreactive horses. The results presented in this study suggest that dogs in Israel could have been accidentally exposed to A. phagocytophilum, for example by ticks carried on migrating birds, however, the possibility of cross‐reaction with E. canis should also be considered. In spite of the high prevalence of ticks on horses in Israel during the summer months, no evidence for exposure to A. phagocytophilum was apparent.
Comparison of Blood Non‐Specific Immune Parameters in Bovine Virus Diarrhoea Virus (BVDV) Persistently Infected and in Immune Heifers
Authors: Piccinini R. , Luzzago C. , Frigerio M. , Frigerio V. , Liandris E. , Zecconi A.
Published: March 2006

Several data from different authors show that Bovine virus diarrhoea virus (BVDV) could be a key component in multiple‐etiology diseases, indeed a lower leukocytes number and their impaired functions decrease the resistance to infections. However, most of the information on the impairment of immune function during BVDV infections arise from circumstantial evidence and from experimental infection studies, and few from field data. To assess the effects of BVDV on blood cells parameters, cellular and humoral functions under field conditions, we designed a controlled study in commercial dairy herds, comparing persistent infected (PI) and healthy heifers. A total of 45 heifers were considered, the PI animals were nine, the control animals were 34, while two controls were considered as acute infected animals. The comparison of the mean values in PI calves showed a significant decrease for leukocytes and granulocytes, while platelets showed a significant increase, when compared with control animals. The total number of lymphocytes decreased not significantly in PI animals, while the proportion significantly increased. The number and proportion of monocytes was significantly reduced in PI animals, when compared with controls. The data collected on markers of cellular immunity during our study cannot be compared with the literature because there are no reference values. The presence of a persistent infection affected the cellular enzymes: NAGase, lysozyme and respiratory burst showed a large statistically significant decrease in PI animals when compared with controls. The presence of a persistent infection with BVD virus influenced blood cells number and impaired some blood cell functions. Such impairment confirms that PI animals represent a threat to the herd not only because they could spread BVDV, but also because they are more susceptible to other infectious diseases.
A Slide Latex Agglutination Test for the Rapid Detection of Antibodies in Serum Against Porcine Parvovirus
Authors: Lü J. , Zhao J. , Fang L. , He Q. , Cao S. , Chen H.
Published: March 2006

A slide latex agglutination test (LAT) was developed and evaluated to detect serum antibodies against porcine parvovirus. Porcine parvovirus antigen was obtained by 10% PEG‐6000 and 0.5 mol/l sodium chloride precipitation, and inactivated by 0.1% methanal. Two per cent suspensions of latex particles (0.5–0.8 μm) were coated by adding an equal volume of porcine parvovirus antigen at 0.34 μg/ml. Repeatability of latex agglutination test was evaluated with a panel of 100 sera using the same and different antigen lots. A good agreement between LAT and haemagglutination inhibit assay was observed. Because of convenience and speed of performance, this method would be used widely in clinic examination.
Detection of the astA (EAST1) Gene in Attaching and Effacing Escherichia coli from Ruminants
Authors: Yuste M. , De La Fuente R. , Ruiz‐santa‐quiteria J. A. , Cid D. , Orden J. A.
Published: March 2006

A total of 206 attaching and effacing Escherichia coli strains from ruminants were analysed for the presence of the astA (EAST1) and bfpA genes. None of these strains was bfpA‐positive. The percentage of enteropathogenic E. coli (EPEC) strains astA‐positive found in healthy cattle (15.6%) suggests that this animal species may be a significant reservoir of atypical EPEC potentially pathogenic for humans.
American foulbrood of the honey bee: Occurrence and distribution of different genotypes of Paenibacillus larvae in the administrative district of Arnsberg (North Rhine‐Westphalia)
Authors: Peters M. , Kilwinski J. , Beringhoff A. , Reckling D. , Genersch E.
Published: March 2006

Between March 2003 and October 2004, Paenibacillus larvae, the aetiological agent of American foulbrood disease of the honey bee, was isolated from broodcombs and honey samples of 54 apiaries in the administrative district of Arnsberg (North Rhine‐Westphalia, Germany). Genotyping of 176 P. larvae isolates with repetitive element polymerase chain reaction fingerprinting (rep‐PCR) using BOX A1R and MBO REP1 primers revealed five different genotypes (AB, Ab, ab, aß, AБ). In samples of three apiaries, more than one genotype was detected. A combination of two genotypes was isolated from honey samples of the same hive two times (ab/aß and Ab/ab). The five genotypes were not randomly distributed in the district, but revealed a certain geographical clustering. Possible factors with impact on the genotype diversity and the distribution pattern are discussed.