bluuebunny/crossref_metadata_2025 huggingface dataset:
Promoter analysis of Helicobacter pylori genes with enhanced expression at low pH
Authors: McGowan Catherine C. , Necheva Antoaneta S. , Forsyth Mark H. , Cover Timothy L. , Blaser Martin J.
Published: May 2003

To identify Helicobacter pylori genes with expression that is enhanced under low pH conditions, we used subtractive hybridization methodology. We identified 28 acid‐induced genes, of which 18 have known or putative functions. Six pairs of genes were co‐transcribed. Primer extension analysis identified single or multiple transcriptional start points (tsp) for 14 of the 22 loci. Sequence analysis of the −10 regions upstream of the tsps revealed consensus motifs for multiple RNA polymerase sigma factors present in H. pylori (σ80, σ54 and σ28). No sequences resembling the −35 Escherichia coli consensus sequence (TTGACA) were present upstream of any of the genes. Both increased gene transcription and decreased mRNA decay contribute to the observed increase in H. pylori transcript abundance at acid pH. These studies document the complex response of H. pylori to environmental pH changes, and provide insight into mechanisms used for intragastric survival.
Role of the Escherichia coli Tat pathway in outer membrane integrity
Authors: Ize Bérengère , Stanley Nicola R. , Buchanan Grant , Palmer Tracy
Published: May 2003

The Escherichia coli Tat system serves to export folded proteins harbouring an N‐terminal twin‐arginine signal peptide across the cytoplasmic membrane. Previous work has demonstrated that strains mutated in genes encoding essential Tat pathway components are highly defective in the integrity of their cell envelope. Here, we report the isolation, by transposon mutagenesis, of tat mutant strains that have their outer membrane integrity restored. This outer membrane repair of the tat mutant arises as a result of upregulation of the amiB gene, which encodes a cell wall amidase. Overexpression of the genes encoding the two additional amidases, amiA and amiC, does not compensate for the outer membrane defect of the tatC strain. Analysis of the amiA and amiC coding sequences indicates that the proteins may be synthesized with plausible twin‐arginine signal sequences, and we demonstrate that they are translocated to the periplasm by the Tat pathway. A Tat+ strain that has mislocalized AmiA and AmiC proteins because of deletion of their signal peptides displays an identical defective cell envelope phenotype. The presence of genes encoding amidases with twin‐arginine signal sequences in the genomes of other Gram‐negative bacteria suggests that a similar cell envelope defect may be a common feature of tat mutant strains.
The Cryptococcus neoformans MAP kinase Mpk1 regulates cell integrity in response to antifungal drugs and loss of calcineurin function
Authors: Kraus Peter R. , Fox Deborah S. , Cox Gary M. , Heitman Joseph
Published: May 2003

Cell wall integrity is crucial for fungal growth, development and stress survival. In the model yeast Saccharomyces cerevisiae, the cell integrity Mpk1/Slt2 MAP kinase and calcineurin pathways monitor cell wall integrity and promote cell wall remodelling under stress conditions. We have identified the Cryptococcus neoformans homologue of the S. cerevisiae Mpk1/Slt2 MAP kinase and have characterized its role in the maintenance of cell integrity in response to elevated growth temperature and in the presence of cell wall synthesis inhibitors. C. neoformans Mpk1 is required for growth at 37°C in vitro, and this growth defect is suppressed by osmotic stabilization. C. neoformans mutants lacking Mpk1 are attenuated for virulence in the mouse model of cryptococcosis. Phosphorylation of Mpk1 is induced in response to perturbations of cell wall biosynthesis by the antifungal drugs nikkomycin Z (a chitin synthase inhibitor), caspofungin (a β‐1,3‐glucan synthase inhibitor), or FK506 (a calcineurin inhibitor), and mutants lacking Mpk1 display enhanced sensitivity to nikkomycin Z and caspofungin. Lastly, we show that calcineurin and Mpk1 play complementing roles in regulating cell integrity in C. neoformans. Our studies demonstrate that pharmacological inhibition of the cell integrity pathway would enhance the activity of antifungal drugs that target the cell wall.
RelE toxins from Bacteria and Archaea cleave mRNAs on translating ribosomes, which are rescued by tmRNA
Authors: Christensen Susanne K. , Gerdes Kenn
Published: May 2003

RelE of Escherichia coli is a global inhibitor of translation that is activated by nutritional stress. Activation of RelE depends on Lon‐mediated degradation of RelB, the antagonist that neutralizes RelE. In vitro, RelE cleaves synthetic mRNAs positioned at the ribosomal A‐site. We show here that in vivo overexpression of RelE confers cleavage of mRNA and tmRNA in their coding regions. RelE‐mediated cleavage depended on translation of the RNAs and occurred at both sense and stop codons. RelE cleavage of mRNA and tmRNA was also induced by amino acid starvation. An ssrA deletion strain was hypersensitive to RelE, whereas overproduction of tmRNA counteracted RelE toxicity. After neutralization of RelE by RelB, rapid recovery of translation required tmRNA, indicating that tmRNA alleviated RelE toxicity by rescuing ribosomes stalled on damaged mRNAs. RelE proteins from Gram‐positive Bacteria and Archaea cleaved tmRNA with a pattern similar to that of E. coli RelE, suggesting that the function and target of RelE may be conserved across the prokaryotic domains.
The DEAD‐box RNA helicase SrmB is involved in the assembly of 50S ribosomal subunits in Escherichia coli
Authors: Charollais Julie , Pflieger Delphine , Vinh Joëlle , Dreyfus Marc , Iost Isabelle
Published: May 2003

Ribosome assembly in Escherichia coli involves 54 ribosomal proteins and three RNAs. Whereas functional subunits can be reconstituted in vitro from the isolated components, this process requires long incubation times and high temperatures compared with the in vivo situation, suggesting that non‐ribosomal factors facilitate assembly in vivo. Here, we show that SrmB, a putative DEAD‐box RNA helicase, is involved in ribosome assembly. The deletion of the srmB gene causes a slow‐growth phenotype at low temperature. Polysome profile analyses of the corresponding cells reveal a deficit in free 50S ribosomal subunits and the accumulation of a new particle sedimenting around 40S. Analysis of the ribosomal RNA and protein contents of the 40S particle indicates that it represents a large subunit that is incompletely assembled. In particular, it lacks L13, one of the five ribosomal proteins that are essential for the early assembly step in vitro. Sucrose gradient fractionation also shows that, in wild‐type cells, SrmB associates with a pre50S particle. From our results, we propose that SrmB is involved in an early step of 50S assembly that is necessary for the binding of L13. This step may consist of a structural rearrangement that, at low temperature, cannot occur without the assistance of this putative RNA helicase.
Surface localized glyceraldehyde‐3‐phosphate dehydrogenase of Mycoplasma genitalium binds mucin
Authors: Alvarez René A. , Blaylock Mark W. , Baseman Joel B.
Published: May 2003

Mycoplasma genitalium is the smallest known self‐replicating cell. It was first isolated from urethral specimens in individuals with non‐gonococcal urethritis and, more recently, from respiratory and synovial sites. Our laboratory has been interested in defining the mechanisms by which M. genitalium adheres to and colonizes host cell surfaces. In order to determine potential targets of adherence, we examined the interaction of M. genitalium with a primary component of the mucosal epithelial lining, mucin (Mn). Three Mn‐binding proteins (MnBPs) of M. genitalium were isolated by affinity chromatography. One of these proteins was identified by N‐terminal sequencing as glyceraldehyde‐3‐phosphate dehy‐drogenase (GAPDH). Antiserum raised against recombinant GAPDH blocked binding of intact biosynthetically labelled mycoplasmas to mucin by approximately 70%. Whole cell radioimmunoprecipitation indicated that GAPDH was surface‐accessible and surface localization of GAPDH was further verified by membrane fractionation and immunoelectron microscopy. The role of GAPDH as an adhesin to Mn not only provides insights into the organism's mechanisms of adherence and colonization but also into its ability to maximize its limited genome.
Proteomic studies of diauxic lag in the differentiating prokaryote Streptomyces coelicolor reveal a regulatory network of stress‐induced proteins and central metabolic enzymes
Authors: Novotna Jana , Vohradsky Jiri , Berndt Peter , Gramajo Hugo , Langen Hanno , Li Xin‐Ming , Minas Wolfgang , Orsaria Lelia , Roeder Daniel , Thompson Charles J.
Published: May 2003

Bacteria typically undergo intermittent periods of starvation and adaptation, emulated as diauxic growth in the laboratory. In association with growth arrest elicited by metabolic stress, the differentiating eubacterium Streptomyces coelicolor not only adapts its primary metabolism, but can also activate developmental programmes leading to morphogenesis and antibiotic biosynthesis. Here, we report combined proteomic and metabolomic data of S. coelicolor used to analyse global changes in gene expression during diauxic growth in a defined liquid medium. Cultures initially grew on glutamate, providing the nitrogen source and feeding carbon (as 2‐oxoglutarate) into the TCA cycle, followed by a diauxic delay allowing reorientation of metabolism and a second round of growth supported by NH4+, formed during prediauxic phase, and maltose, a glycolytic substrate. Cultures finally entered stationary phase as a result of nitrogen starvation. These four physiological states had previously been defined statistically by their distinct patterns of protein synthesis and heat shock responses. Together, these data demonstrated that the rates of synthesis of heat shock proteins are determined not only by temperature increase but also by the patterns and rates of metabolic flux in certain pathways. Synthesis profiles for metabolic‐ and stress‐induced proteins can now be interpreted by the identification of 204 spots (SWICZ database presented at http:proteom.biomed.cas.cz). Cluster analysis showed that the activity of central metabolic enzymes involved in glycolysis, the TCA cycle, starvation or proteolysis each displayed identifiable patterns of synthesis that logically underlie the metabolic state of the culture. Diauxic lag was accompanied by a structured regulatory programme involving the sequential activation of heat‐, salt‐, cold‐ and bacteriostatic antibiotic (pristinamycin I, PI)‐induced stimulons. Although stress stimulons presumably provide protection during environmental‐ or starvation‐induced stress, their identities did not reveal any coherent adaptive or developmental functions. These studies revealed interactive regulation of metabolic and stress response systems including some proteins known to support developmental programmes in S. coelicolor.
Substrate preference of stress‐activated phospholipase D in Chlamydomonas and its contribution to PA formation
Authors: Arisz Steven A. , Valianpour Fredoen , Van Gennip Albert H. , Munnik Teun
Published: May 2003

In response to various environmental stress conditions, plants rapidly form the intracellular lipid second messenger phosphatidic acid (PA). It can be generated by two independent signalling pathways via phospholipase D (PLD) and via phospholipase C (PLC) in combination with diacylglycerol kinase (DGK). In the green alga Chlamydomonas, the phospholipid substrates for these pathways are characterized by specific fatty acid compositions. This allowed us to establish: (i) PLD's in vivo substrate preference; and (ii) PLD's contribution to PA formation during stress signalling. Accordingly, G‐protein activation (1 µm mastoparan), hyperosmotic stress (150 mm NaCl) and membrane depolarization (50 mm KCl) were used to stimulate PLD, as monitored by the accumulation in 5 min of its unique transphosphatidylation product phosphatidylbutanol (PBut). In each case, PBut's fatty acid composition specifically matched that of phosphatidylethanolamine (PE), identifying this lipid as PLD's favoured substrate. This conclusion was substantiated by analysing the molecular species by electrospray ionization‐mass spectrometry (ESI‐MS/MS), which revealed that PE and NaCl‐induced PBut share a unique (18 : 1)2‐structure. The fatty acid composition of PA was much more complex, reflecting the different contributions from the PLC/DGK and PLD pathways. During KCl‐induced stress, the PA rise was largely accounted for by PLD activity. In contrast, PLD's contribution to hyperosmotic stress‐induced PA was less, being approximately 63% of the total increase. This was because the PLC/DGK pathway was activated as well, resulting in phosphoinositide‐specific fatty acids and molecular species in PA.
Sequence‐based alignment of sorghum chromosome 3 and rice chromosome 1 reveals extensive conservation of gene order and one major chromosomal rearrangement
Authors: Klein Patricia E. , Klein Robert R. , Vrebalov Julia , Mullet John E.
Published: May 2003

The completed rice genome sequence will accelerate progress on the identification and functional classification of biologically important genes and serve as an invaluable resource for the comparative analysis of grass genomes. In this study, methods were developed for sequence‐based alignment of sorghum and rice chromosomes and for refining the sorghum genetic/physical map based on the rice genome sequence. A framework of 135 BAC contigs spanning approximately 33 Mbp was anchored to sorghum chromosome 3. A limited number of sequences were collected from 118 of the BACs and subjected to blastx analysis to identify putative genes and blastn analysis to identify sequence matches to the rice genome. Extensive conservation of gene content and order between sorghum chromosome 3 and the homeologous rice chromosome 1 was observed. One large‐scale rearrangement was detected involving the inversion of an approximately 59 cM block of the short arm of sorghum chromosome 3. Several small‐scale changes in gene collinearity were detected, indicating that single genes and/or small clusters of genes have moved since the divergence of sorghum and rice. Additionally, the alignment of the sorghum physical map to the rice genome sequence allowed sequence‐assisted assembly of an approximately 1.6 Mbp sorghum BAC contig. This streamlined approach to high‐resolution genome alignment and map building will yield important information about the relationships between rice and sorghum genes and genomic segments and ultimately enhance our understanding of cereal genome structure and evolution.
The anticodon and the D‐domain sequences are essential determinants for plant cytosolic tRNAVal import into mitochondria
Authors: Delage Ludovic , Duchêne Anne‐Marie , Zaepfel Marlyse , Maréchal‐Drouard Laurence
Published: May 2003

In higher plants, one‐third to one‐half of the mitochondrial tRNAs are encoded in the nucleus and are imported into mitochondria. This process appears to be highly specific for some tRNAs, but the factors that interact with tRNAs before and/or during import, as well as the signals present on the tRNAs, still need to be identified. The rare experiments performed so far suggest that, besides the probable implication of aminoacyl‐tRNA synthetases, at least one additional import factor and/or structural features shared by imported tRNAs must be involved in plant mitochondrial tRNA import. To look for determinants that direct tRNA import into higher plant mitochondria, we have transformed BY2 tobacco cells with Arabidopsis thaliana cytosolic tRNAVal(AAC) carrying various mutations. The nucleotide replacements introduced in this naturally imported tRNA correspond to the anticodon and/or D‐domain of the non‐imported cytosolic tRNAMet–e. Unlike the wild‐type tRNAVal(AAC), a mutant tRNAVal carrying a methionine CAU anticodon that switches the aminoacylation of this tRNA from valine to methionine is not present in the mitochondrial fraction. Furthermore, mutant tRNAsVal carrying the D‐domain of the tRNAMet–e, although still efficiently recognized by the valyl‐tRNA synthetase, are not imported any more into mitochondria. These data demonstrate that in plants, besides identity elements required for the recognition by the cognate aminoacyl‐tRNA synthetase, tRNA molecules contain other determinants that are essential for mitochondrial import selectivity. Indeed, this suggests that the tRNA import mechanism occurring in plant mitochondria may be different from what has been described so far in yeast or in protozoa.
A C‐terminal sequence of soybean β‐conglycinin α′ subunit acts as a vacuolar sorting determinant in seed cells
Authors: Nishizawa Keito , Maruyama Nobuyuki , Satoh Ryohei , Fuchikami Yoshihiro , Higasa Takahiko , Utsumi Shigeru
Published: May 2003

In maturing seed cells, many newly synthesized proteins are transported to the protein storage vacuoles (PSVs) via vesicles unique to seed cells. Vacuolar sorting determinants (VSDs) in most of these proteins have been determined using leaf, root or suspension‐cultured cells apart from seed cells. In this study, we examined the VSD of the α′ subunit of β‐conglycinin (7S globulin), one of the major seed storage proteins of soybean, using Arabidopsis and soybean seeds. The wild‐type α′ was transported to the matrix of the PSVs in seed cells of transgenic Arabidopsis, and it formed crystalloid‐like structures. Some of the wild‐type α′ was also transported to the translucent compartments (TLCs) in the PSV presumed to be the globoid compartments. However, a derivative lacking the C‐terminal 10 amino acids was not transported to the PSV matrix, and was secreted out of the cells, although a portion was also transported to the TLCs. The C‐terminal region of α′ was sufficient to transport a green fluorescent protein (GFP) to the PSV matrix. These indicate that α′ contains two VSDs: one is present in the C‐terminal 10 amino acids and is for the PSV matrix; and the other is for the TLC (the globoid compartment). We further verified that the C‐terminal 10 amino acids were sufficient to transport GFP to the PSV matrix in soybean seed cells by using a transient expression system.
A polyglycine stretch is necessary for proper targeting of the protein translocation channel precursor to the outer envelope membrane of chloroplasts
Authors: Inoue Kentaro , Keegstra Kenneth
Published: May 2003

Toc75 is a protein translocation channel in the outer envelope membrane of chloroplasts and its presence is essential for the biogenesis of the organelles. Toc75 is the only protein identified so far in the outer membrane of chloroplasts or mitochondria that is synthesized as a larger precursor, preToc75, with a bipartite transit peptide. Its N‐terminus targets the protein to the stroma and is removed by the stromal processing peptidase, whereas its C‐terminus mediates envelope targeting and is removed by a yet unknown peptidase. Several conserved domains have been identified in the C‐terminal portion of the preToc75 transit peptide from six plant species. We evaluated their importance in the biogenesis of Toc75 by means of deletion or site‐directed mutagenesis, followed by import experiments using isolated chlroplasts. Among the conserved domains, a polyglycine stretch was found to be necessary for envelope targeting. Substitution of this domain with other stretches of a single amino acid such as alanine caused mistargeting of the protein into the stroma, indicating an important role for this domain. Furthermore, a glutamate at +2 and two alanine residues at −3 and −1 to the second cleavage site were found to be important for processing. A potential mechanism for the biogenesis of Toc75 is discussed.
Activation tagging, a novel tool to dissect the functions of a gene family
Authors: Nakazawa Miki , Ichikawa Takanari , Ishikawa Akie , Kobayashi Hiroko , Tsuhara Yumi , Kawashima Mika , Suzuki Kumiko , Muto Shu , Matsui Minami
Published: May 2003

In a screen for morphological mutants from the T1 generation of approximately 50 000 activation‐tagging lines, we isolated four dominant mutants that showed hyponastic leaves, downward‐pointing flowers and decreased apical dominance. We designated them isoginchaku (iso). The iso‐1D and iso‐2D are allelic mutants caused by activation of the AS2 gene. The T‐DNAs were inserted in the 3′ downstream region of AS2. Iso‐3D and iso‐4D are the other allelic mutants caused by activation of the ASL1/LBD36 gene. These two genes belong to the AS2 family that is composed of 42 genes in Arabidopsis. The only recessive mutation isolated from this gene family was of AS2, which resulted in a leaf morphology mutant. Applying reverse genetics using a database of activation‐tagged T‐DNA flanking sequences, we found a dominant mutant that we designated peacock1‐D (pck1‐D) in which the ASL5/LBD12 gene was activated by a T‐DNA. The pck1‐D mutants have lost apical dominance, have epinastic leaves and are sterile. These results strongly suggest that activation tagging is a powerful mutant‐mining tool especially for genes that make up a gene family.
Host‐related variation among isolates of the Sirococcus shoot blight pathogen from conifers
Authors: Smith D. R. , Bronson J. J. , Stanosz G. R.
Published: May 2003

Variation among isolates of the Sirococcus shoot blight pathogen (attributed to Sirococcus conigenus, Sirococcus sp., or one of many synonyms) was studied. Inter‐simple‐sequence‐repeat‐anchored polymerase chain reaction fingerprints, nuclear rDNA internal transcribed spacers and 5.8S sequences, and 18S rDNA sequences were analysed. In addition, culture morphology as well as sizes and shapes of conidia produced on seed cone scales in culture were compared. Two main groups were distinguished based on both genetic markers and cultural characteristics. The P group consists of isolates mostly from pine and spruce from Europe and both eastern and western North America; the T group consists of isolates mostly from hemlock from western North America. Results provide a benchmark for comparison of other isolates of these fungi and indicate the need for additional sampling and comparative research of these pathogens.
First records of soilborne Phytophthora species in Swedish oak forests
Authors: Jönsson U. , Lundberg L. , Sonesson K. , Jung T.
Published: May 2003

Thirty‐two oak stands in southern Sweden, 27 with predominantly declining trees and five with a higher proportion of healthy trees were investigated regarding the presence of soilborne Phytophthora species. Phytophthora quercina, an oak‐specific fine root pathogen, was isolated from rhizosphere soil samples in 10 of the 27 declining stands. Additionally, P. cactorum and P. cambivora were recovered from one stand each. No Phytophthora species were isolated from the healthy oak stands. The soil conditions at the sites from which Phytophthora spp. were recovered ranged from mesic sediments to moraines, with clayey to silty textures and with soil pH (BaCl2) between 3.5 and 5.0. The results show that P. quercina is geographically widespread in oak stands in southern Sweden and indicate that this pathogen may be one of the factors involved in oak decline in Northern Europe as has already been shown for western, Central and parts of southern Europe.
Resinous stem canker development during the growing season of Chamaecyparis obtusa (Hinoki cypress) inoculated with pathogenic fungus Cistella japonica
Authors: Yamada T. , Hasegawa E. , Miyashita S.‐I.
Published: May 2003

Seasonal development of resinous stem canker was determined in Chamaecyparis obtusa trees wound‐inoculated with Cistella japonica in January 1996. Samples for anatomical analysis were harvested on 20 May, 9 July and 6 August in the second year of inoculation, and the sections were observed under light microscope. Resin exudation was most abundant in the second year, decreasing in the third. All new resin exudation in the second year was recognized at the May observation. New traumatic resin canal formation was observed in the August samples, and an expansion of necrotic lesions was observed mainly in summer samples in the inoculated trees and in naturally affected trees. Cistella japonica was re‐isolated from all bark lesions or inoculated wounds examined. These results suggest that the activity of Ci. japonica in the tree affects the seasonal development of resinous stem canker during the growing season of the trees.
The Results of Using Faecal Culture as Confirmation Test of Paratuberculosis‐seropositive Dairy Cattle
Authors: Muskens J. , Mars M. H. , Elbers A. R. W. , Van Maanen K. , Bakker D. , Kaaden O.‐R.
Published: May 2003

A total of 15 822 cattle aged 3 years and older, belonging to 378 randomly selected herds, were tested for paratuberculosis using an absorbed enzyme‐linked immunosorbent assay (ELISA); 3.3% tested positive. This percentage was lowest for the group of cattle aged 3–4 years (2.3%) and highest for cattle with the age of 5–6 years (4.5%). The mean Sample to Positive (S/P) ratio of seropositive cattle vaccinated against paratuberculosis was higher (0.75 ± 0.33) than that of seropositive, non‐vaccinated cattle (0.58 ± 0.26). Faecal samples of 422 ELISA‐positive cattle were cultured for the presence ofMycobacterium aviumsubsp.paratuberculosis, 12% of these were contaminated. The percentage of non‐contaminated samples with positive culture results was 17.3%, with a substantial difference between vaccinated (1.7%) and non‐vaccinated cattle (20.2%). Of the positive cultures, the number of colonies varied from 1–10 (22% of cultures), 11–100 (22%), to more than 100 (55%). The percentage of ELISA‐positive, non‐vaccinated cattle tested culture‐positive was positively correlated with the magnitude of the S/P ratio. This percentage varied from 12% (S/P ratio 0.3–0.5) to 58% (S/P ratio > 1.1), a result that might have implications for interpretation of the test. In this study, the percentage of ELISA‐positive cattle with positive faecal culture results was limited and these individuals were mostly moderate to heavy shedders.
Apoptosis in the Cerebellum of Dogs with Distemper
Authors: Moro L. , Martins A. S. , Alves C. M. , Santos F. G. A. , Del Puerto H. L. , Vasconcelos A. C.
Published: May 2003

Canine distemper virus (CDV) may induce multifocal demyelination in the central nervous system of infected dogs. The pathogenesis of this process is not clear. The present work identifies the presence of apoptotic cells in white and grey matter of dogs’cerebellum, naturally infected with CDV. Fifteen dogs with clinical signs of canine distemper that tested positive for CDV nucleoprotein were used. Brain specimens were processed and embedded in paraffin. Sections 5 μm thick were stained with hematoxylin–eosin and Shorr. Other sections were submitted to TUNEL reaction and to immunohistochemistry for CDV nucleoprotein detection. Acute and chronic demyelinated plaques were observed in the white matter, while apoptosis occurred particularly in the granular layer of grey matter. Apoptosis seems to play an important role in the pathogenesis of canine distemper demyelination.
Prevalence of Antibodies to Borrelia burgdorferi sensu lato in Southern Chamois (Rupicapra pyrenaica) in Spain
Authors: Ortuño A. , Castellà J. , Marco I. , Ruiz M. , Lavín S. , Kaaden O.‐R.
Published: May 2003

Ungulates are involved in the epidemiology of Borreliosis as maintenance hosts for vector tick species. We evaluated the prevalence of antibodies to Borrelia burgdorferi sensu lato in two different populations of southern chamois in Spain. Forty‐six sera (40.4%) were positive at titres ≥1 : 64 using an indirect immunofluorescence assay (IFAT).
Differentiation of Staphylococcus sciuri Strains Isolated from Free‐Living Rodents and Insectivores
Authors: Hauschild T. , Schwarz S.
Published: May 2003

Twenty‐nine Staphylococcus sciuri strains isolated from free‐living insectivores and rodents were comparatively analysed for their biochemical capacities and their SmaI macrorestriction patterns. The 29 S. sciuri isolates represented 21 different biotypes and 22 different SmaI macrorestriction types. This observation confirmed that S. sciuri isolates obtained from insectivores and rodents living in natural environments constituted a heterogeneous population. Cluster analysis revealed that the macrorestriction patterns and the biochemical profiles matched in some cases. However, S. sciuri isolates that exhibited the same or closely related biochemical profiles were also found to be associated with different macrorestriction patterns. Analysis of the 29 S. sciuri isolates for their plasmid content and their sensitivity to antimicrobial agents showed that most of the isolates were plasmid‐free and susceptible to all antimicrobial agents tested.
Virulence of Recent and Former Classical Swine Fever Virus Isolates Evaluated by their Clinical and Pathological Signs
Authors: Floegel‐Niesmann G. , Bunzenthal C. , Fischer S. , Moennig V. , Kaaden O.‐R.
Published: May 2003

The clinical diagnosis of classical swine fever (CSF) still caused problems to the veterinarians during the last decade. The primary CSF outbreak was often detected too late and, meanwhile, the virus had spread. Consequently, the recent classical swine fever virus isolates (CSFV) were suspected to be of low virulence. The purpose of the study was to quantify the virulence of four recent CSFV by evaluating the clinical and pathological signs caused by different CSFV. Pigs of the same breed and age group were inoculated intranasally with CSFV from recent epidemics in European Union (EU) member states. The CSFV used are registered in the data base of the EU Reference Laboratory for CSF and belong to different genotypes: 2.1, 2.2 and 2.3 respectively. Clinical signs of CSF were evaluated by using a score system suggested previously (Mittelholzer et al., 2000: Vet. Microbiol. 74, 293). For the evaluation of pathological lesions, a new pathological score was introduced. The four CSFV tested here were classified as moderately virulent in general, although one CSFV may cause different clinical courses, ranging from highly virulent to avirulent. This indicates the importance of additional factors in the host animal for virulence. Differences in the clinical and pathological signs between these four recent CSFV were rather minor, emphasizing that the genetic typing of CSFV is absolutely essential. Differences towards former CSFV (e.g. reference virus strain Alfort  187) were more pronounced, especially regarding the onset and duration of the disease, the occurrence of skin haemorrhages and pathological lesions of kidney, subcutis and serosae. It is concluded that clinical diagnosis of CSF is rather difficult in pigs up to 14 days post‐CSFV infection using these four CSFV, emphasizing the need for careful differential diagnosis and the laboratory investigation for CSF at an early stage.
Coinfection by Cryptosporidium parvum and Porcine Circovirus Type 2 in Weaned Pigs
Authors: Núñez A. , McNeilly F. , Perea A. , Sánchez‐Cordón P. J. , Huerta B. , Allan G. , Carrasco L.
Published: May 2003

Routine histopathological diagnosis of one representative 3‐month‐old pig from a group suffering from diarrhoea revealed a massive degree of parasitation by Cryptosporidium parvum, with a concomitant infection by porcine circovirus type 2 (PCV2), that was confirmed by immunohistochemical procedures. The areas of intestine where parasites were more numerous presented abundant PCV2 infected cells in mucosa and submucosa. The concurrence of C. parvum, a rare primary intestinal pathogen in post‐weaning and growing pigs, and PCV2 infections suggest an increased susceptibility as a result of an immunosuppression state.
Eradication of Aujeszky's Disease in Germany
Authors: Müller T. , Bätza H.‐J. , Schlüter H. , Conraths F. J. , Mettenleiter T. C.
Published: May 2003

Aujeszky's disease (AD) manifested itself in both German states in 1960. Owing to the historical development, in the subsequent two decades, the development of the disease and of its control in the Western and Eastern parts of Germany went different ways. This article describes differences and particularities in the development of AD in Germany leading to the establishment of a national AD eradication programme after re‐unification of the two German states at the beginning of the last decade. The basic principles of the German AD eradication programme are described, and the results of 10 years of efforts to control the disease are presented and discussed. Without any doubt, as in other European countries, implementation of the national eradication programme resulted in a considerable progress in the eradication of AD. Since the eradication programme has been established in 1989, particularly in West Germany, the number of AD outbreaks has decreased steadily from about 2000 cases in 1987 to 0 cases recorded in 2001. Recently, Germany has been declared as officially AD‐free by the European Commission.
Validity and Psychometric Properties of the General Health Questionnaire-12 in Young Australian Adolescents
Authors: Tait Robert J , French Davina J , Hulse Gary K
Published: May 2003

Background: The General Health Questionnaire (GHQ) is a measure of current mental wellbeing that has been extensively validated with adults. The instrument has also been used with adolescents.

Objective: (i) To assess the psychometric properties of the GHQ-12 among school students in grades 7–10; (ii) to validate it against other psychological tests; and (iii) to suggest a threshold score.

Method: The survey was conducted in single sex and mixed schools from the state and private system in Perth, Western Australia. The survey contained the GHQ-12 and measures of anxiety, depression, self-esteem, stress, generalized self-efficacy, social desirability and negative affectivity.

Results: There were 336 students (female 55%) with an age range of 11–15 years (median 13). The GHQ showed good internal consistency (alpha 0.88). Girls had higher mean GHQ-12 scores than boys (F (1,326) 15.0, p < 0.001) and scores for both genders increased with school grade (F (3,326) 4.2, p < 0.01). Multiple linear regression showed that depression, anxiety, self-esteem and stress were significant independent predictors of GHQ scores. The model accounted for 68% of the variance (adjusted R 2). Screening indices were calculated by comparison with a combined depression and/or anxiety category. Threshold scores of 13/14 for males and 18/19 for females appeared optimal. General Health Questionnaire scores were compared with two criterion groups: adolescents in hospital with alcohol or drug (AOD) related problems and those with problems not related to AOD use. Only the former group had significantly higher total scores.

Conclusions: The GHQ-12 showed good structural characteristics and was appropriately correlated with other measures of related traits. Overall, the GHQ-12 appears to be a valid index of psychological wellbeing in this population and was considerably shorter than some of the other instruments.
Randomized Controlled Trials in Psychiatry. Part 1: Methodology and Critical Evaluation
Authors: Porter Richard , Frampton Chris , Joyce Peter R. , Mulder Roger T.
Published: May 2003

Objective: To introduce clinicians to the methodology and critical appraisal of randomized controlled trials (RCTs) in psychiatry.

Method: The methodology of RCTs in psychiatry is discussed. Using a systematic approach to critical appraisal, a published RCT of treatments for severe depression is examined and evaluated.

Results and Conclusions: The RCT appraised illustrates certain problematic areas in the methodology of trials in psychiatry. A detailed knowledge of methodology and critique of RCTs is essential in determining whether reported results will influence clinicians’ practice.
Moral Responsibility and Borderline Personality Disorder
Authors: Bray Amanda
Published: May 2003

Objective: To examine the concepts of free will and moral responsibility in a psychiatric context, and to consider whether those with borderline personality disorder (BPD) may be considered responsible for their actions.

Method: A wide, but necessarily incomplete, range of literature was reviewed in the fields of psychiatry and philosophy. I offer a set of criteria for responsible action and examine some features of BPD in the light of these criteria.

Results: Impulsivity, acting out and the less severe forms of dissociation do not vitiate responsibility. Severe dissociative and psychotic symptoms may well render people with BPD less morally responsible for their actions.

Conclusions: Comorbid conditions in BPD may also affect the ability to act responsibly.
Substance Use Disorder Comorbidity Among Inpatient Youths With Psychiatric Disorder
Authors: Swadi Harith , Bobier Candace
Published: May 2003

Objective: Substance abuse/dependence has been reported to show significant association with psychopathology, and is likely to influence the course and outcome of psychiatric illness. The aim of this study was to determine the rate of substance use disorders (other than alcohol) comorbidity among inpatient adolescents with severe Axis 1 psychiatric disorder.

Method: A retrospective analysis of systematically collected data was carried out. The subjects were 16–18-year-old youths, admitted to an inpatient unit for severe psychiatric disorder. The data collection process utilized the DSM-IV criteria for diagnostic categorization of psychiatric disorder and substance use disorder. Demographic data, and data on suicide attempts were also collected.

Results: Over a period of one year (March 2001–March 2002), 62 patients were admitted to the Christchurch Youth Inpatient Unit; 40 (64.5%) had a comorbid Substance Abuse Disorder (SAD) according to DSM-IV criteria and none had a Substance Dependence Disorder. The vast majority involved cannabis and stimulants. Sixty per cent of those with mood disorder, 63% of those with anxiety disorder and 80% of those with schizophrenia spectrum disorder had a comorbid SAD. Internalizing problems, especially mood disorders, predominated among those with SAD reflecting the Unit's admission criteria. There were no differences in attempted suicide rates between those with SAD and those without SAD, but those with SAD were more likely to have unstable accommodation/living arrangements than those without SAD.

Conclusions: Our findings confirm previous reports suggestive of high rates of SAD comorbidity among youth with severe psychiatric illness. There are clinical and process implications for these findings particularly identification of substance use disorders and their treatment as well as resource availability and staff training.
TRAVELLERS: A School-Based Early Intervention Programme Helping Young People Manage and Process Change, Loss and Transition. Pilot Phase Findings
Authors: Dickinson Pauline , Coggan Carolyn , Bennett Sara
Published: May 2003

Objective: This paper outlines the conceptual background and findings from the pilot phase of TRAVELLERS – an early intervention programme designed to enhance protective factors for young people experiencing change, loss and transition events and early signs of emotional distress. The pilot study aimed to determine whether TRAVELLERS was a feasible, acceptable and promising intervention for young people within secondary schools in Aotearoa/New Zealand.

Method: The conceptual origins of the TRAVELLERS programme are described in terms of: adolescent mental health concerns; emerging mental health promotion theory and practice; and prevention and early intervention models. The key elements of the TRAVELLERS programme are described. The programme was piloted in two secondary schools, one rural and one urban with 34 participants (females n = 24, males n = 10). Evaluation methods included: review of programme materials; identification of potential selection tools appropriate to Year 9 students; analysis of selection questionnaire; and conduct of feedback from participants, facilitators and parents/caregivers.

Results: The TRAVELLERS programme provides a means of identifying and selecting young people who may benefit from participating in an early intervention programme. The programme has achieved a statistically significant reduction in participants’ distress (p < 0.01). Young people were overwhelmingly enthusiastic about most aspects of TRAVELLERS. School personnel reported that TRAVELLERS was an appropriate and acceptable programme to the school.

Conclusions: Targeted interventions provided within a supportive school environment can contribute to enhancing protective factors such as personal and interpersonal coping strategies, increased help-seeking behaviour, and young people feeling more positive about themselves and their lives. The pilot programme has been amended and prepared for a two year trial phase in 10 secondary schools during 2002–2003.
Bouffée Délirante and ICD-10 Acute and Transient Psychoses: A Comparative Study
Authors: Pillmann Frank , Haring Annette , Balzuweit Sabine , Blöink Raffaela , Marneros Andreas
Published: May 2003

Objective: ICD-10 acute and transient psychotic disorder (ATPD; F23) has one of its historical roots in the French concept of bouffée délirante. This study explores the empirical relationship of the two concepts.

Method: During a 5-year period, all inpatients with ATPD were identified; the diagnosis of bouffée délirante was also determined. We systematically evaluated demographic and clinical features and carried out follow-up investigations at an average of 2.2 years after the index episode using standardized instruments.

Results: Forty-two (4.1%) of 1036 patients treated for psychotic disorders or major affective episode fulfilled the ICD-10 criteria of ATPD. Of these, only 28.6% also fulfilled the criteria of bouffée délirante. Patients with bouffée délirante were significantly younger than the remaining ATPD patients. Outcome parameters were generally more favourable for patients diagnosed with bouffée délirante than for ATPD patients without a concurrent diagnosis of bouffée délirante, but the difference was statistically significant only for occupational status.

Conclusions: There are indications that a diagnosis of bouffée délirante carries a somewhat better prognosis than ATPD in general. However, the low frequency of operationally diagnosed bouffée délirante suggests that the criteria might be too narrow.
Awareness of Illness in Patients With Bipolar I Disorder With or Without Comorbid Anxiety Disorders
Authors: Pini Stefano , Dell'osso Liliana , Amador Xavier F. , Mastrocinque Concettina , Saettoni Marco , Cassano Giovanni B.
Published: May 2003

Background: The present study examined whether specific types of comorbid anxiety disorders, namely panic disorder (PD), social phobia (SP) and obsessive–compulsive disorder (OCD) are differentially associated with course variables and insight into bipolar illness.

Method: The sample consisted of 151 consecutively hospitalized patients with bipolar I disorder. They were assessed in the week prior to discharge using the Structured Clinical Interview for DSM-III-R (SCID-P), the Brief Psychiatric Rating Scale (BPRS), the Global Assessment of Functioning Scale (GAF) and the Hopkins Symptom Checklist (HSCL-90). Level of insight was assessed with the Scale to assess Unawareness of Mental Disorders (SUMD).

Results: Of the 151 bipolar subjects, 92 had no PD, SP and OCD comorbidity, 35 had PD and 24 had SP and/or OCD. The three groups differed significantly on the current awareness of illness and treatment response scores and the retrospective awareness of illness and treatment response scores. Post-hoc analyses revealed that, compared with bipolar patients without PD/SD/OCD and those with comorbid PD, patients with comorbid SP and/or OCD had better insight on current awareness of illness, current awareness of treatment response, retrospective awareness of illness and retrospective awareness of treatment response. The regression analysis showed that the presence of no panic type anxiety comorbidity was a predictor of good insight.

Conclusions: These data indicate the value of identifying comorbid anxiety disorders in patients with bipolar illness. The results could be interpreted as evidence of discrete disorders within the bipolar spectrum, one that is characterized by, among other things, SP and/or OCD with good insight, another characterized by PD with poor insight.
Evaluation of parathyroid hyperplasia by ultrasonographic examination in patients with end‐stage renal failure before and at initiation of dialysis
Authors: HAMADA CHIEKO , FUKUI MITSUMINE , SAKAMOTO TOMOKO , KOIZUMI MICHIKO , ISHIGURO CHIZURU , OSADA SHIWORI , SHOU ICHIYU , HAYASHI KAYO , TOMINO YASUHIKO
Published: May 2003

SUMMARY:  Secondary hyperparathyroidism (2HPT), which is related to renal osteodystrophy (ROD), may occur in patients in the comparatively early stage of chronic renal failure (CRF). Secondary hyperparathyroidism patients with parathyroid hyperplasia showed resistance to vitamin D3 treatment during long‐term dialysis. At present, evaluation by ultrasonography is considered to be useful for confirming parathyroid hyperplasia. There are no clinical data associated with imaging evaluation of 2HPT in CRF patients. In the present study, the relationship among clinical and biochemical data, and parathyroid hyperplasia by ultrasonography, was evaluated in 12 patients (six males and six females) with end‐stage renal failure (ESRF) before and at initiation of dialysis. Five patients showed an enlargement of parathyroid glands in ultrasonography. Levels of serum‐intact parathyroid hormone (PTH) in patients with parathyroid hyperplasia (positive group) were significantly higher than in those without hyperplasia (negative group; 97.6 ± 36.65 vs 17.4 ± 4.45 pmol/L; P < 0.05). The levels of intact PTH were above 35.0 pmol/L in all five patients with hyperplasia. All patients in the positive group had never taken vitamin D3 supplements. Calcium‐containing phosphate binders were not prescribed before the present study, except in one patient. Parathyroid hyperplasia caused by 2HPT was recognized in patients before and at initiation of dialysis in this study. It appears that untreated 2HPT in CRF patients may progress to advanced 2HPT in ESRF before and/or after the early stage of dialysis. The levels of serum intact PTH are useful in predicting parathyroid hyperplasia.
Clinical observations of mycophenolate mofetil therapy in refractory primary nephrotic syndrome
Authors: ZHAO MINGHUI , CHEN XIANGMEI , CHEN YIPU , LIU ZHANGSUO , LIU YUCHUN , LU FANGPING , ZHANG YANPING , WANG HAIYAN
Published: May 2003

SUMMARY:  Mycophenolate mofetil (MMF) is an effective immunosuppressive agent in renal transplantation, and preliminary studies suggest that it may also be effective in the treatment of lupus nephritis. This study investigated the efficacy and safety of MMF therapy in patients with refractory primary nephrotic syndrome in a prospective multicentre clinical observation. Nineteen refractory nephrotic patients with minimal change disease or mesangial proliferative glomerulonephritis were enrolled in this study. Combined MMF and prednisone therapy was used for 6 months with an initial MMF dose of 1.0–2.0 g/day and a prednisone dose of 20–60 mg/day; both drugs were tapered gradually. It was found that all patients achieved clinical remission and 11 of 19 responded within 4 weeks, and 12 of 19 patients entered complete clinical remission. The prednisone dose in those patients who were previously steroid dependent could be successfully tapered. During follow up, three patients experienced transient increasing of proteinuria associated with infections and recovered without an adjustment of therapy. One patient was withdrawn from the study because of a fall in haemoglobin levels; other adverse effects did not necessitate withdrawal. Follow‐up renal biopsies in two patients found no alteration in renal pathology. Mycophenolate mofetil is an effective and well‐tolerated immunosuppressive agent for patients with refractory nephrotic syndrome.