bluuebunny/crossref_metadata_2025 huggingface dataset:
Comprehensive Comparison of Auxin-Regulated and Brassinosteroid-Regulated Genes in Arabidopsis
Authors: Goda Hideki , Sawa Shinichiro , Asami Tadao , Fujioka Shozo , Shimada Yukihisa , Yoshida Shigeo
Published: April 2004

Although numerous physiological studies have addressed the interactions between brassinosteroids and auxins, little is known about the underlying molecular mechanisms. Using an Affymetrix GeneChip representing approximately 8,300 Arabidopsis genes, we studied comprehensive transcript profiles over 24 h in response to indole-3-acetic acid (IAA) and brassinolide (BL). We identified 409 genes as BL inducible, 276 genes as IAA inducible, and 637 genes in total. These two hormones regulated only 48 genes in common, suggesting that most of the actions of each hormone are mediated by gene expression that is unique to each. IAA-up-regulated genes were enriched in genes regulated in common. They were induced quickly by IAA and more slowly by BL, suggesting divergent physiological roles. Many were early auxin-inducible genes and their homologs, namely SAUR, GH3, and IAA. The comprehensive comparison also identified IAA- and BL-specific genes, which should help to elucidate the specific actions of each hormone. The identified genes were classified using hierarchical clustering based on the similarity of their responses to the two hormones. Gene classification also allowed us to analyze the frequency of cis-elements. The TGTCTC element, a core element of the previously reported auxin response element, was not enriched in genes specifically regulated by IAA but was enriched in the 5′-flanking region of genes up-regulated by both IAA and BL. Such gene classification should be useful for predicting the functions of unknown genes, to understand the roles of these two hormones, and the promoter analysis should provide insight into the interaction of transcriptional regulation by the two hormones.
Protein Cryoprotective Activity of a Cytosolic Small Heat Shock Protein That Accumulates Constitutively in Chestnut Stems and Is Up-Regulated by Low and High Temperatures
Authors: Lopez-Matas Maria-Angeles , Nuñez Paulina , Soto Alvaro , Allona Isabel , Casado Rosa , Collada Carmen , Guevara Maria-Angeles , Aragoncillo Cipriano , Gomez Luis
Published: April 2004

Heat shock, and other stresses that cause protein misfolding and aggregation, trigger the accumulation of heat shock proteins (HSPs) in virtually all organisms. Among the HSPs of higher plants, those belonging to the small HSP (sHSP) family remain the least characterized in functional terms. We analyzed the occurrence of sHSPs in vegetative organs of Castanea sativa (sweet chestnut), a temperate woody species that exhibits remarkable freezing tolerance. A constitutive sHSP subject to seasonal periodic changes of abundance was immunodetected in stems. This protein was identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry and internal peptide sequencing as CsHSP17.5, a cytosolic class I sHSP previously described in cotyledons. Expression of the corresponding gene in stems was confirmed through cDNA cloning and reverse transcription-PCR. Stem protein and mRNA profiles indicated that CsHSP17.5 is significantly up-regulated in spring and fall, reaching maximal levels in late summer and, especially, in winter. In addition, cold exposure was found to quickly activate shsp gene expression in both stems and roots of chestnut seedlings kept in growth chambers. Our main finding is that purified CsHSP17.5 is very effective in protecting the cold-labile enzyme lactate dehydrogenase from freeze-induced inactivation (on a molar basis, CsHSP17.5 is about 400 times more effective as cryoprotectant than hen egg-white lysozyme). Consistent with these observations, repeated freezing/thawing did not affect appreciably the chaperone activity of diluted CsHSP17.5 nor its ability to form dodecameric complexes in vitro. Taken together, these results substantiate the hypothesis that sHSPs can play relevant roles in the acquisition of freezing tolerance.
Cloning, Functional Expression, and Characterization of the Raffinose Oligosaccharide Chain Elongation Enzyme, Galactan:Galactan Galactosyltransferase, from Common Bugle Leaves
Authors: Tapernoux-Lüthi Esther M. , Böhm Andreas , Keller Felix
Published: April 2004

Galactan:galactan galactosyltransferase (GGT) is a unique enzyme of the raffinose family oligosaccharide (RFO) biosynthetic pathway. It catalyzes the chain elongation of RFOs without using galactinol (α-galactosyl-myoinositol) by simply transferring a terminal α-galactosyl residue from one RFO molecule to another one. Here, we report the cloning and functional expression of a cDNA encoding GGT from leaves of the common bugle (Ajuga reptans), a winter-hardy long-chain RFO-storing Lamiaceae. The cDNA comprises an open reading frame of 1215 bp. Expression in tobacco (Nicotiana plumbaginifolia) protoplasts resulted in a functional recombinant protein, which showed GGT activity like the previously described purified, native GGT enzyme. At the amino acid level, GGT shows high homologies (>60%) to acid plant α-galactosidases of the family 27 of glycosylhydrolases. It is clearly distinct from the family 36 of glycosylhydrolases, which harbor galactinol-dependent raffinose and stachyose synthases as well as alkaline α-galactosidases. Physiological studies on the role of GGT confirmed that GGT plays a key role in RFO chain elongation and carbon storage. When excised leaves were exposed to chilling temperatures, levels of GGT transcripts, enzyme activities, and long-chain RFO concentrations increased concomitantly. On a whole-plant level, chilling temperatures induced GGT expression mainly in the roots and fully developed leaves, both known RFO storage organs of the common bugle, indicating an adaptation of the metabolism from active growth to transient storage in the cold.
The Effect of α-Amanitin on the Arabidopsis Seed Proteome Highlights the Distinct Roles of Stored and Neosynthesized mRNAs during Germination
Authors: Rajjou Loïc , Gallardo Karine , Debeaujon Isabelle , Vandekerckhove Joël , Job Claudette , Job Dominique
Published: April 2004

To investigate the role of stored and neosynthesized mRNAs in seed germination, we examined the effect of α-amanitin, a transcriptional inhibitor targeting RNA polymerase II, on the germination of nondormant Arabidopsis seeds. We used transparent testa mutants, of which seed coat is highly permeable, to better ascertain that the drug can reach the embryo during seed imbibition. Even with the most permeable mutant (tt2-1), germination (radicle protrusion) occurred in the absence of transcription, while subsequent seedling growth was blocked. In contrast, germination was abolished in the presence of the translational inhibitor cycloheximide. Taken together, the results highlight the role of stored proteins and mRNAs for germination in Arabidopsis and show that in this species the potential for germination is largely programmed during the seed maturation process. The α-amanitin-resistant germination exhibited characteristic features. First, this germination was strongly slowed down, indicating that de novo transcription normally allows the synthesis of factor(s) activating the germination rate. Second, the sensitivity of germination to gibberellic acid was reduced 15-fold, confirming the role of this phytohormone in germination. Third, de novo synthesis of enzymes involved in reserve mobilization and resumption of metabolic activity was repressed, thus accounting for the failure in seedling establishment. Fourth, germinating seeds can recapitulate at least part of the seed maturation program, being capable of using mRNAs stored during development. Thus, commitment to germination and plant growth requires transcription of genes allowing the imbibed seed to discriminate between mRNAs to be utilized in germination and those to be destroyed.
Partial Purification, Kinetic Analysis, and Amino Acid Sequence Information of a Flavonol 3-O-Methyltransferase from Serratula tinctoria
Authors: Huang Tyng-Shyan , Anzellotti Dominique , Dedaldechamp Fabienne , Ibrahim Ragai K.
Published: April 2004

Serratula tinctoria (Asteraceae) accumulates mainly 3,3′-dimethylquercetin and small amounts of 3-methylquercetin as an intermediate. The fact that 3-methylquercetin rarely accumulates in plants in significant amounts, and given its important role as an antiviral and antiinflammatory agent that accumulates in response to stress conditions, prompted us to purify and characterize the enzyme involved in its methylation. The flavonol 3-O-methyltransferase (3-OMT) was partially purified by ammonium sulfate precipitation and successive chromatography on Superose-12, Mono-Q, and adenosine-agarose affinity columns, resulting in a 194-fold increase of its specific activity. The enzyme protein exhibited an expressed specificity for the methylation of position 3 of the flavonol, quercetin, although it also utilized kaempferol, myricetin, and some monomethyl flavonols as substrates. It exhibited a pH optimum of 7.6, a pI of 6.0, and an apparent molecular mass of 31 kD. Its K  m values for quercetin as the substrate and S-adenosyl-l-Met (AdoMet) as the cosubstrate were 12 and 45 μ  m, respectively. The 3-OMT had no requirement for Mg2+, but was severely inhibited by p-chloromercuribenzoate, suggesting the requirement for SH groups for catalytic activity. Quercetin methylation was competitively inhibited by S-adenosyl-l-homo-Cys with respect to the cosubstrate AdoMet, and followed a sequential bi-bi reaction mechanism, where AdoMet was the first to bind and S-adenosyl-l-homo-Cys was released last. In-gel trypsin digestion of the purified protein yielded several peptides, two of which exhibited strong amino acid sequence homology, upon protein identification, to a number of previously identified Group II plant OMTs. The availability of peptide sequences will allow the design of specific nucleotide probes for future cloning of the gene encoding this novel enzyme for its use in metabolic engineering.
Tissue-Specific Localization of an Abscisic Acid Biosynthetic Enzyme, AAO3, in Arabidopsis
Authors: Koiwai Hanae , Nakaminami Kentaro , Seo Mitsunori , Mitsuhashi Wataru , Toyomasu Tomonobu , Koshiba Tomokazu
Published: April 2004

Arabidopsis aldehyde oxidase 3 (AAO3) is an enzyme involved in abscisic acid (ABA) biosynthesis in response to drought stress. Since the enzyme catalyzes the last step of the pathway, ABA production sites may be determined by the presence of AAO3. Here, AAO3 localization was investigated using AAO3 promoter:AAO3-GFP transgenic plants and by an immunohistochemical technique. AAO3-GFP protein exhibited an activity to produce ABA from abscisic aldehyde, and the transgene restored the wilty phenotype of the aao3 mutant. GFP-fluorescence was detected in the root tips, vascular bundles of roots, hypocotyls and inflorescence stems, and along the leaf veins. Intense immunofluorescence signals were localized in phloem companion cells and xylem parenchyma cells. Faint but significant GFP- and immuno-fluorescence signals were observed in the leaf guard cells. In situ hybridization with antisense AAO3 mRNA showed AAO3 mRNA expression in the guard cells of dehydrated leaves. These results indicate that the ABA synthesized in vascular systems is transported to various target tissues and cells, and also that the guard cells themselves are able to synthesize ABA.
Potato Virus X-Induced Gene Silencing in Leaves and Tubers of Potato
Authors: Faivre-Rampant Odile , Gilroy Eleanor M. , Hrubikova Katarina , Hein Ingo , Millam Steve , Loake Gary J. , Birch Paul , Taylor Mark , Lacomme Christophe
Published: April 2004

Virus induced gene silencing (VIGS) is increasingly used to generate transient loss-of-function assays and has potential as a powerful reverse-genetics tool in functional genomic programs as a more rapid alternative to stable transformation. A previously described potato virus X (PVX) VIGS vector has been shown to trigger silencing in the permissive host Nicotiana benthamiana. This paper demonstrates that a PVX-based VIGS vector is also effective in triggering a VIGS response in both diploid and cultivated tetraploid Solanum species. We show that systemic silencing of a phytoene desaturase gene is observed and maintained throughout the foliar tissues of potato plants and was also observed in tubers. Here we report that VIGS can be triggered and sustained on in vitro micropropagated tetraploid potato for several cycles and on in vitro generated microtubers. This approach will facilitate large-scale functional analysis of potato expressed sequence tags and provide a noninvasive reverse-genetic approach to study mechanisms involved in tuber and microtuber development.
TOUSLED Kinase Activity Oscillates during the Cell Cycle and Interacts with Chromatin Regulators
Authors: Ehsan Hashimul , Reichheld Jean-Philippe , Durfee Tim , Roe Judith L.
Published: April 2004

The TOUSLED (TSL)-like nuclear protein kinase family is highly conserved in plants and animals. tsl loss of function mutations cause pleiotropic defects in both leaf and flower development, and growth and initiation of floral organ primordia is abnormal, suggesting that basic cellular processes are affected. TSL is more highly expressed in exponentially growing Arabidopsis culture cells than in stationary, nondividing cells. While its expression remains constant throughout the cell cycle in dividing cells, TSL kinase activity is higher in enriched late G2/M-phase and G1-phase populations of Arabidopsis suspension culture cells compared to those in S-phase. tsl mutants also display an aberrant pattern and increased expression levels of the mitotic cyclin gene CycB1;1, suggesting that TSL represses CycB1;1 expression at certain times during development or that cells are delayed in mitosis. TSL interacts with and phosphorylates one of two Arabidopsis homologs of the nucleosome assembly/silencing protein Asf1 and histone H3, as in humans, and a novel plant SANT/myb-domain protein, TKI1, suggesting that TSL plays a role in chromatin metabolism.
Functional Analysis of the Final Steps of the 1-Deoxy-d-xylulose 5-phosphate (DXP) Pathway to Isoprenoids in Plants Using Virus-Induced Gene Silencing
Authors: Page Jonathan E. , Hause Gerd , Raschke Maja , Gao Wenyun , Schmidt Jürgen , Zenk Meinhart H. , Kutchan Toni M.
Published: April 2004

Isoprenoid biosynthesis in plant plastids occurs via the 1-deoxy-d-xylulose 5-phosphate (DXP) pathway. We used tobacco rattle virus (TRV) to posttranscriptionally silence the expression of the last two enzymes of this pathway, the IspG-encoded (E)-4-hydroxy-3-methylbut-2-enyl diphosphate synthase (HDS) and the IspH-encoded isopentenyl/dimethylallyl diphosphate synthase (IDDS), as well as isopentenyl/dimethylallyl diphosphate isomerase (IDI), the enzyme that interconverts IPP and DMAPP. TRV-IspG and TRV-IspH infected Nicotiana benthamiana plants had albino leaves that contained less than 4% of the chlorophyll and carotenoid pigments of control leaves. We applied [13C]DXP and [14C]DXP to silenced leaves and found that 2-C-methyl-d-erythritol 2,4-cyclodiphosphate accumulated in plants blocked at HDS while DXP, (E)-4-hydroxy-3-methylbut-2-enyl phosphate and (E)-2-methylbut-2-ene-1,4-diol accumulated in IDDS-blocked plants. Albino leaves from IspG- and IspH-silenced plants displayed a disorganized palisade mesophyll, reduced cuticle, fewer plastids, and disrupted thylakoid membranes. These findings demonstrate the participation of HDS and IDDS in the DXP pathway in plants, and support the view that plastid isoprenoid biosynthesis is metabolically and physically segregated from the mevalonate pathway. IDI-silenced plants had mottled white-pale green leaves with disrupted tissue and plastid structure, and showed an 80% reduction in pigments compared to controls. IPP pyrophosphatase activity was higher in chloroplasts isolated from IDI-silenced plants than in control plant chloroplasts. We suggest that a low level of isoprenoid biosynthesis via the DXP pathway can occur without IDI but that this enzyme is required for full function of the DXP pathway.
Adenylate Gradients and Ar:O2 Effects on Legume Nodules. II. Changes in the Subcellular Adenylate Pools
Authors: Wei Hui , Atkins Craig A. , Layzell David B.
Published: April 2004

Central infected zone tissue of soybean (Glycine max L. Merr.) nodules was fractionated into separate subcellular compartments using density gradient centrifugation in nonaqueous solvents to better understand how exposure to Ar:O2 (80:20%, v/v) atmosphere affects C and N metabolism, and to explore a potential role for adenylates in regulating O2 diffusion. When nodules were switched from air to Ar:O2, adenylate energy charge (AEC) in the plant cytosol rose from 0.63 ± 0.02 to 0.73 ± 0.02 within 7 min and to 0.80 ± 0.01 by 60 min. In contrast, AEC of the mitochondrial compartment of this central zone tissue remained high (0.80 ± 0.02 to 0.81 ± 0.02) following Ar treatment while that of the bacteroid compartment was unchanged, at 0.73 ± 0.02, after 7 min, but declined to 0.57 ± 0.03 after 60 min. These results were consistent with a simulation model that predicted Ar:O2 exposure would first reduce ATP demand for ammonia assimilation and rapidly increase cytosolic AEC, before the Ar:O2-induced decline mediated by a decrease in nodule O2 permeability reduces bacteroid AEC. The possibility that adenylates play a key, integrating role in regulating nodule permeability to oxygen diffusion is discussed.
Anthropology as art, art as anthropology: Tate Modern, London
Authors: Bowman Glenn
Published: April 2004

Conference reviewed:

Fieldworks: Dialogues between art and anthropology: Tate Modern, London, 26–28 September 2003
Agency, Discourses of Power and Collective Representations: Socrates
Authors: Streissler Anna
Published: April 2004

Conference reviewed:

Socrates Intensive Programme 2003, University Of Vienna, 26 August–4 September 2003
Skin Diseases in Chinese Children at a Pediatric Dermatology Center
Authors: Hon Kam Lun , Leung Ting Fan , Wong Yin , Ma Kwok Chiu , Fok Tai Fai
Published: April 2004

Abstract:  Understanding the pattern of disease referrals is important in the delivery of a service. We followed 331 new referrals at a pediatric dermatology clinic over 12 months. Fifty percent of the patients were female; 293 patients (88.5%) had a single diagnosis and 36 (10.9%) had two diagnoses. Three leading causes for referrals accounted for 60% of the 371 skin diseases encountered: 124 diagnoses (33%) were of eczema, 73 diagnoses (20%) were of nevi, and 22 diagnoses (6%) were of viral warts. The nevi seen included melanocytic, epidermal, sebaceous, and vascular nevi. Skin biopsies were performed in 23 patients (6.9%), and microscopy and culture for fungal infections in 11 patients (3.3%). Forty‐one patients (12%) were referred to the laser clinic for assessment of their nevi or pigmented skin lesions. Topical steroids were prescribed in 47% of patients during their follow‐ups. Follow‐up appointments were offered to 90% of patients. Eczema required the most frequent follow‐up. Gender disparity in referrals was evident only in endogenous eczema (odds ratio [OR] 1.64, 95% confidence interval [CI] 1.04–2.60 in males; p = 0.033). Girls were more likely to be referred to the laser clinic than boys (OR 2.31, 95% CI 1.10–4.92; p = 0.026). Most dermatologic diagnoses were made on clinical grounds and moderately potent topical corticosteroids were the most commonly prescribed medication. Since chronic skin disorders prevailed in the referrals, repeated follow‐up appointments were required.
Rhinoscleroma in Three Siblings
Authors: Fernández Vozmediano J. M. , Armario Hita J. C. , Cabrerizo A. González
Published: April 2004

Abstract:  Rhinoscleroma is a chronic, granulomatous infectious disease that responds poorly to treatment. In recent years an increasing number of cases have been reported in nonendemic areas, explained largely by major migratory movements. We describe rhinoscleroma in three siblings. They had ulcerated but painless lesions, which bled spontaneously, and hemorrhagic scabs or crusts in their noses. In one child, the lesions had destroyed the entire left nasal ala and alar cartilage and most of the right. Dermatopathologic study identified the Mikulicz macrophages that contained organisms. It is possible that disposing factors could have been the neutropenia common to the three children and their poor living conditions. They were treated with a combination of trimethoprim‐sulfamethoxazole and cefalexin, for a period of 3 months. We present this unusual case history of three siblings affected by a process that is relatively infrequent in our area of practice and is not considered very contagious. It is important to recognize the clinical signs characteristic of this disease, the diagnosis of which is not easy. Improvements in living conditions, hygiene, and health standards are essential prerequisites for its control and prevention.
Bathtub Suction‐Induced Purpura
Authors: Landers Maeran C. , Schroeder Theresa L.
Published: April 2004

Abstract:  We report two cases of bathtub suction‐induced purpura. Two girls, ages 9 and 14 years, developed recurring purpura “spontaneously” in the same lower back location and in the same U‐shaped distribution. The patients had similar histopathologic findings, with extravasated erythrocytes and a sparse superficial inflammatory infiltrate. After extensive evaluation, one patient eventually admitted to self‐inducing her lesions by creating suction against a bathtub. The second patient denied self‐inflicting her lesions; however, her purpura improved when she stopped taking baths.
Infantile Systemic Hyalinosis or Juvenile Hyaline Fibromatosis?
Authors: Urbina Francisco , Sazunic Ivo , Murray Guillermo
Published: April 2004

Abstract:  Infantile systemic hyalinosis and juvenile hyaline fibromatosis are presumably autosomal recessive inherited diseases of unknown origin in which accumulation of an amorphous, hyaline material occurs in the skin and other organs. Both disorders may show clinical overlapping, suggesting that they might represent different variants of the same disease spectrum. We describe a 6‐year‐old boy with such overlap. Salient features included papular skin lesions on his face and neck, gingival hyperplasia, perianal nodules, large subcutaneous tumors on the scalp, hyperpigmented plaques over the metacarpophalangeal joints and malleoli, limited joint movement, diffuse osteopenia, short stature, and persistent diarrhea. Histopathologic and ultrastructural studies confirmed the presence of hyalin material in the dermis. The term systemic hyalinosis involves both conditions and should be preferred until a clear distinction can be made between them.
Chloroplast development affects expression of phage‐type RNA polymerases in barley leaves
Authors: Emanuel Carola , Weihe Andreas , Graner Andreas , Hess Wolfgang R. , Börner Thomas
Published: April 2004

We have identified the barley gene and cDNA encoding the plastid phage‐type RNA polymerase (RNAP), nuclear‐encoded plastid RNAP (RpoTp), and the nearly full‐length cDNA of the mitochondrial RNAP, nuclear‐encoded mitochondrial RNAP (RpoTm). RpoTp spans more than 9000 nt, consists of 19 exons and 18 introns, gives rise to a 3632‐nt mRNA and is localized to the long arm of chromosome 1 (7H). The length of the deduced polypeptide is 948 residues. The mRNA levels of RpoTp and RpoTm were determined in roots and primary leaf sections of 7‐day‐old barley seedlings of the albostrians mutant, which were either phenotypically normal and exhibited a gradient of chloroplast development, or contained ribosome‐deficient undifferentiated plastids. Transcript levels of RpoTp and RpoTm in almost all sections reached higher concentrations in plastid ribosome‐deficient leaves than in the wild‐type material, except in the most basal part of the leaf. These data indicate a role of plastid‐to‐nucleus signalling in the expression of the two RpoT genes. The mRNA levels of the plastid genes, β‐subunit of plastid‐encoded RNAP (rpoB), proteolytic subunit of the Clp protease (clpP) and ribosomal protein Rpl2 (rpl2), all known to be transcribed by the nuclear‐encoded RNAP (NEP), followed closely the pattern of RpoTp mRNA accumulation, strongly suggesting that RpoTp and NEP are identical. Transcripts of RpoTm and RpoTm‐transcribed mitochondrial genes cytochrome oxidase subunit 2 (coxII) and ATPase subunit 9 (atp9) accumulated to the highest levels in the most basal parts of the leaf and declined considerably towards the leaf tip with a pronounced reduction in green versus white leaves. Our data revealed a marked influence of the developmental stage of the plastid on the expression and activity of organellar phage‐type RNAPs and their target genes. Thus, interorganellar cross‐talk in the regulated expression of nuclear‐encoded plastid and mitochondrial RNAP genes might be a key element governing the concerted expression of genes located within plastids, mitochondria and the nucleus of the plant cell.
Optimizing wound treatment through health care structuring and professional education
Authors: Gottrup Finn
Published: April 2004

Considerable developments have occurred in wound healing and care over the last few decades. Innovative technologies such as use of growth factors, bioengineered skin, topical negative pressure, heat, oxygen, and others have entered the clinic. These techniques have been advantageous, but even the most advanced and sophisticated product requires proper wound care and wound bed preparation in order to function optimally. It has been shown that the organization of a wound team, not the change in products, increases the healing of problem wounds (MTV Report, Denmark, 2003). Efficacy studies are generally performed in a few centers with investigators already experienced in the field. This probably leads to an optimal efficacy of the product. Problems arise when the product is marketed and is used by clinicians not specially educated in wound care. If no treatment plan and education is performed, product efficacy and cost‐effectiveness decrease dramatically. This is clearly a major problem for patients, but also for communities and, in the end, for companies' sale of the product. Structuring of treatment and education is for these reasons the largest present challenge in the area of wound healing. The optimal way to treat nonhealing wounds is based on a multidisciplinary concept that has been implemented in a national health care system as a recognized expert function. It should consist of a specific treatment plan including modern wound care products and well‐educated personnel. This structure is similar to other areas of the medical profession, but has to a lesser degree been established in the treatment of problem wounds. A growing understanding of the importance of this area is, however, shown by the increasing number of working groups focusing on organization and education in wound care. Presently the European Wound Management Association, the World Union of Wound Healing Societies, and The Danish Wound Healing Society have created such types of working groups.
Recombinant factor VIIa prophylaxis in a patient with severe congenital factor VII deficiency
Authors: Tcheng W. Y. , Donkin J. , Konzal S. , Wong W.‐Y.
Published: April 2004

Summary.  Use of recombinant factor VIIa (rFVIIa, NovoSeven®) in patients with congenital FVII deficiency has been reported for the prophylactic management of surgical bleeding and for the treatment of acute bleeding episodes. Because of its short half‐life, the use of rFVIIa on a regular prophylactic regimen has not been routinely adopted. In this report, we describe our successful experience with rFVIIa prophylaxis in preventing recurrent target joint bleeding in a severely FVII‐deficient adolescent.
Is there a ‘Basque’ profile regarding autosomal recessive deficiencies of coagulation factors?*
Authors: Bauduer F. , Ducout L. , Dutour O. , Degioanni A.
Published: April 2004

Summary.  When excluding haemophilia and von Willebrand disease, coagulation factors deficiencies constitute rare autosomal recessive disorders (<1 in 500 000) of less precisely defined epidemiology. We have reported herein the distribution of these entities in the French Basque Country, a genetic isolate of very old individualization with peculiar biological specificities. The prevalence of these disorders was markedly high, especially, as already shown, factor XI deficiency. This unusual profile needs to be discussed in the view of population genetics.
Laboratory diagnosis of von Willebrand's disorder: quality and diagnostic improvements driven by peer review in a multilaboratory test process
Authors: Favaloro E. J. , Bonar R. , Kershaw G. , Sioufi J. , Hertzberg M. , Street A. , Lloyd J. , Marsden K. ,   on behalf of the RCPA QAP in Haematology
Published: April 2004

Summary.  Regular multilaboratory surveys of laboratories derived primarily from Australia, New Zealand and Southeast Asia have been conducted over the past 7 years to evaluate testing proficiency in the diagnosis of von Willebrand's disorder (VWD) and to assess changes to test practice. Participating laboratories (currently 45) are asked to perform their usual panel of tests for VWD, and then to self‐interpret test results as to the likelihood (or not) of VWD, as well as to the potential subtype identified. Samples provided in the past two survey distributions (both conducted in 2003) were as follows. Survey part A/distribution 1: Normal donor plasma, plasma with borderline normal/reduced levels of VWF (×2) and plasma from an individual with type 2 A VWD. Survey part B/distribution 2 (family VWD study): Plasma from a father, mother and son with borderline normal/reduced von Willebrand factor (VWF), and a daughter with type 3 VWD. In line with previously published survey results, the interassay and within method coefficients of variation (CV) were similar for all assays (around 15–25%), although tending to be slightly higher for VWF:RCo and VWF:CB than VWF:Ag and FVIII:C. Most laboratories reported test values consistent with expected findings, and made correct interpretations or predictions regarding the nature of the samples, although discrepant assay results or interpretations are still seen in approximately 5–10% of responses (typically from laboratories using a more limited test panel or not performing the VWF:CB). Overall, problems with the non‐identification of functional VWF discordance in type 2 VWD, the misidentification of functional VWF discordance in type 1 VWD, and difficulties in discriminating types 1 and 3 VWD appear to predominate. In comparison with previous surveys, performance of electro‐immuno diffusion (EID) (or Laurel gel) procedures has now ceased, and a reduction in VWF:RCo and VWF:Multimer testing and an increase in latex immunoassay (LIA) testing is sustained. We conclude that laboratories are generally proficient in tests for VWD, and that diagnostic error rates are reduced when test panels are more comprehensive and include the VWF:CB.
Substance P and other neuropeptides do not induce mediator release in isolated human intestinal mast cells
Authors: Bischoff S. C. , Schwengberg S. , Lorentz A. , Manns M. P. , Bektas H. , Sann H. , Levi‐Schaffer F. , Shanahan F. , Schemann M.
Published: April 2004

Abstract  Neuropeptides such as substance P (SP) and related peptides are supposed to act as mast cell agonists, and thus as mediators of neuroimmune interactions. The data supporting this hypothesis were obtained mostly from rodent experiments. Here, we studied for the first time the effect of SP and other peptides on mediator release in human intestinal mast cells, either unpurified or enriched to 85–99% purity. We found that SP at 0.1–100 μmol L−1, or other peptides including neurokinin A and B, calcitonin gene‐related peptide, vasoactive intestinal peptide and serotonin at 1 μmol L−1 do not induce release of mediators such as histamine, sulphidoleukotrienes, and tumour necrosis factor α. The peptides also failed to cause mediator release in mast cells isolated from inflamed tissue derived from Crohn's disease. Using reverse transcriptase‐polymerase chain reaction, flow cytometry and immunohistochemistry, we could show that human intestinal mast cells do not express the tachykinin receptors NK‐1, NK‐2, or NK‐3 under basal conditions. However, upon stimulation by immunoglobulin E (IgE) receptor‐crosslinking, which induces an extensive mediator release reaction, a subpopulation of mast cells clearly expressed NK‐1, the SP receptor. In conclusion, our data show that SP and other neuropeptides do not act as secretagogues in human intestinal mast cells that have not been pre‐activated by IgE receptor‐crosslinking.
Effects of clonidine and tricyclic antidepressants on gastric smooth muscle contractility
Authors: James A. N. , Ryan J. P. , Parkman H. P.
Published: April 2004

Abstract  To determine if and how clonidine and tricyclic antidepressants affect gastric contractility. Guinea pig fundic and antral circular muscle strips were studied in vitro. The effects of clonidine or amitriptyline added in graded concentrations on contractions to electric field stimulation (EFS), acetylcholine (ACh), and SP in the presence of N‐nitro‐l‐arginine methyl ester (l‐NAME) were studied. EFS produced frequency dependent contractions of fundic and antral muscle that were abolished by atropine or tetrodotoxin (TTX). ACh contractions were abolished by atropine but not TTX. Clonidine reduced contractile response to EFS but had no effect on ACh contractions. The threshold concentration of clonidine to inhibit EFS contractions was lower in the fundus than in the antrum. Amitriptyline reduced contractions to both EFS and ACh but not to SP. The threshold concentration of amitriptyline to inhibit EFS contractions was lower in the antrum than in the fundus. Both clonidine and amitriptyline affect gastric contractility. At threshold concentrations, clonidine affects fundic contractility whereas amitriptyline affects antral contractility. Clonidine affects gastric contractility in response to EFS but not to ACh, suggesting alpha‐2 receptors on cholinergic nerves that reduce ACh release. Amitriptyline inhibits gastric contractility to EFS and ACh suggesting an inhibitory muscle effect.
Pathophysiology as a basis for understanding symptom complexes and therapeutic targets
Authors: Camilleri M. , Talley N. J.
Published: April 2004

Abstract  Sensorimotor disorders of the stomach, small intestine and colon have a limited repertoire of clinical manifestations, and there is the potential for more than one mechanism to lead to symptoms. In many recent clinical trial programs of novel agents in neurogastroenterology, the emphasis has been primarily on symptom assessment of broad groups of patients identified by the Rome criteria. Drugs of potential value have fallen by the wayside with this approach. We propose the current paradigm is partly to blame; physiological testing should provide the basis for identifying more homogeneous populations and therapeutic targets within functional bowel disease, and this applies to the upper and lower gut. Here we summarize the evidence that certain biomarkers can, in a limited fashion, be used to predict the success of an experimental medicine in common disorders of gastrointestinal function, including the irritable bowel syndrome and functional dyspepsia. Although the current evidence is limited and is most convincingly demonstrated with examples of transit measurements (for loperamide, alosetron, tegaserod and piboserod), we perceive this paradigm that studies using validated and responsive biomarkers have an important role to play in drug development.
Oxidative burst and expression of germin/oxo genes during wounding of ryegrass leaf blades: comparison with senescence of leaf sheaths
Authors: Le Deunff Erwan , Davoine Céline , Le Dantec Christelle , Billard Jean‐Pierre , Huault Claude
Published: April 2004

Two bursts of H2O2 production have been detected by in situ 3,3′‐diaminobenzidine (DAB) staining after cutting of Lolium perenne L. leaf blades. The first burst, which occurred immediately after wounding was inhibited by Na‐diethydithiocarbamate (DIECA), a Cu/Zn–superoxide dismutase (SOD) inhibitor. The second burst, which was initiated several hours later, coincided with the induction of oxalate oxidase (G‐OXO) activity detected in vitro or visualized in situ by the α‐chloronaphtol assay. Four genes encoding G‐OXO have been identified from cDNA obtained from wounded L. perenne L. leaf blades. Comparison of protein sequences revealed more than 91% homology in the coding region between G‐OXOs of the true cereals and G‐OXOs of ryegrass, which is a Gramineae belonging to the tribe of Festucaceae. The wound‐dependent increase of G‐OXO activity in floated cut leaf blades was the result of differential induction of the four g‐oxo genes. The involvement of G‐OXOs in wound‐induced H2O2 production coincided with the presence in leaf tissues of oxalate throughout the period of increase of G‐OXO synthesis. Moreover, expression of g‐oxo genes was enhanced by an exogenous supply of H2O2 or methyljasmonate (MeJa). Expression of the four g‐oxo genes was also induced after in planta stinging of leaf blades. The pattern of their expression in planta was identical to that occuring in senescing leaf sheaths. These results emphasize the importance of G‐OXOs in H2O2 production in oxalate‐producing plant species such as ryegrass. G‐OXOs might be crucial during critical events in the life of plants such as cutting and senescence by initiating H2O2‐mediated defences against pathogens and foraging animals.
A mutation of the CRUMPLED LEAF gene that encodes a protein localized in the outer envelope membrane of plastids affects the pattern of cell division, cell differentiation, and plastid division in Arabidopsis
Authors: Asano Tomoya , Yoshioka Yasushi , Kurei Shunsuke , Sakamoto Wataru ,   Sodmergen, Machida Yasunori
Published: April 2004

We identified a novel mutation of a nuclear‐encoded gene, designated as CRUMPLED LEAF (CRL), of Arabidopsis thaliana that affects the morphogenesis of all plant organs and division of plastids. Histological analysis revealed that planes of cell division were distorted in shoot apical meristems (SAMs), root tips, and embryos in plants that possess the crl mutation. Furthermore, we observed that differentiation patterns of cortex and endodermis cells in inflorescence stems and root endodermis cells were disturbed in the crl mutant. These results suggest that morphological abnormalities observed in the crl mutant were because of aberrant cell division and differentiation. In addition, cells of the crl mutant contained a reduced number of enlarged plastids, indicating that the division of plastids was inhibited in the crl. The CRL gene encodes a novel protein with a molecular mass of 30 kDa that is localized in the plastid envelope. The CRL protein is conserved in various plant species, including a fern, and in cyanobacteria, but not in other organisms. These data suggest that the CRL protein is required for plastid division, and it also plays an important role in cell differentiation and the regulation of the cell division plane in plants. A possible function of the CRL protein is discussed.
National pension systems and mass opinion: a case study of confidence, satisfaction and political attitudes in Norway
Authors: Bay Ann‐Helén , Pedersen Axel West
Published: April 2004

It is commonly assumed that popular support for national pension systems depends on widespread satisfaction with projected benefit levels among the working age population, and in particular that public support for the system will be jeopardised if the taxpayers do not feel confident about eventually receiving the promised benefits. On the basis of Norwegian survey data, two sets of questions are addressed in the article: (1) Is there a widespread lack of confidence in and satisfaction with the Norwegian National Insurance pension scheme? and (2) Is there an association between confidence and satisfaction and people's political attitudes towards the National Insurance pension scheme? Although we do not find any signs of a dramatic erosion of confidence towards the system, we do find that overall satisfaction with projected benefits is low among the working age population. Contrary to what one might expect, however, confidence and satisfaction from the point of view of individual interests appear not to be associated with a political preference for privatisation.
A Malar Granular Cell Tumor in a Djungarian Hamster
Authors: Sato T. , Takada M. , Shibuya H. , Shirai W. , Matsuo K.
Published: April 2004

‘Granular cell’ tumor observed in the malar subcutis of a Djungarian hamster was examined to determine its cellular origin. Histologically, the tumor consisted of a solid growth of oval or spindle‐shaped large cells with abundant cytoplasm filled with eosinophilic granules that were periodic acid‐Schiff‐positive and diastase‐resistant. Immunohistochemically, the tumor cells were positive to anti‐vimentin and anti‐desmin antibodies and a few cells showed positivity to anti‐actin antibody as well. They did not react to myoglobin, S‐100 protein, and glial fibrillary acidic protein (GFAP). Electron microscopic studies revealed that the tumor cells had pinocytotic vesicles, dense plaque and microfilaments. The first granular cell variant of myogenic tumor reported here in Djungarian hamsters was differentiated from granular cell tumor of Schwann cell origin.
Pregnancy Rate after Timed Artificial Insemination in Early Post‐partum Dairy Cows after Ovsynch or Specific Synchronization Protocols
Authors: Lòpez‐Gatius F. , Murugavel K. , Santolaria P. , López‐Béjar M. , Yániz J. L.
Published: April 2004

The present study was designed to compare the reproductive performance of pre‐synchronized post‐partum dairy cows subjected, either to the Ovsynch protocol without screening for ovarian status (control group), or to a specific oestrous synchronization protocol applied according to their ovarian status, as determined by transrectal ultrasound (experimental group). The study was conducted on 428 lactating dairy cows. Cows in the Ovsynch group (n = 205) were synchronized and time inseminated after receiving the Ovsynch protocol treatment. Cows in the specific synchronization (Ssynch) group (n = 223) were weekly subjected to transrectal ultrasound exams for 4 weeks, or until AI or starting treatment, and divided into four subgroups according to their ovarian status: (i) corpus luteum (CL) subgroup (n = 130), cows with a CL; (ii) natural oestrus (NE) subgroup (n = 58), cows showing NE; (iii) anovulatory follicles (AF) subgroup (n = 26), cows considered to have AF; and (iv) ovarian cysts (OC) subgroup (n = 9), cows with OC. Cows in the Ssynch group were synchronized and time inseminated following a specific oestrous synchronization protocol, or inseminated at NE. Logistic regression analysis was carried out for the dependent variables ovulation and pregnancy rates to first and to second AI (second AI: first AI + return AI). Cows subjected to Ssynch were 2.1 times more likely to become pregnant at first and at second AI compared with those synchronized using the Ovsynch protocol (P < 0.0001). Our results show that the response of post‐partum pre‐synchronized cows to a specific oestrous synchronization protocol applied according to their ovarian status is more effective than the response to the Ovsynch protocol applied without taking into account the ovarian status of the animals.
Immunohistopathological Findings in the Lungs of Calves Naturally Infected with Mycoplasma bovis
Authors: Khodakaram‐Tafti A. , López A.
Published: April 2004

Histology and immunohistochemistry were used to analyse the lesions and distribution of Mycoplasma bovis antigen in the lungs of 18 naturally infected calves. Microscopic examination of pneumonic lungs revealed two distinct patterns of necrosis and inflammation. The first pattern was observed in six of 18 (33.3%) calves in which microscopic lesions were characterized by large irregular areas of coagulative necrosis surrounded by a dense zone of degenerated neutrophils. Moderate amounts of mycoplasmal antigen were in the centre and periphery of these necrotic foci and, to a lesser extent, in mononuclear cells of the peribronchial lymphoid tissue. The second pattern was observed in 18 of 18 (100%) calves and consisted of rounded foci of caseous necrosis composed by granular eosinophilic material surrounded by a rim of granulation tissue. Large amounts of M. bovis antigen were detected in the centre and periphery of these necrotic foci and, to a lesser extent, in the peribronchial lymphoid tissue, and alveolar and interstitial macrophages. It was concluded that both caseous and coagulative necrosis of the lung parenchyma was primarily caused by M. bovis. Infection with M. bovis should be suspected in bovine necrotic bronchopneumonia, particularly in cases in which the pulmonary necrosis is part of a pyogranulomatous inflammation centred around airways. The pattern of caseous necrosis with pyogranulomatous inflammation is characteristic of M. bovis infection while the pattern of coagulative necrosis is similar to and must be differentiated from Mannheimia haemolytica and Haemophilus somnus infection.
Clinical and Pathological Features of the Porcine Ulcerative Dermatitis Syndrome (PUDS)
Authors: Schmoll F. , Sipos W. , Kahlbacher H. , Schilcher F. , Bagó Z. , Bunka S. , Miller I. , Schuh M.
Published: April 2004

The nomenclature of ulcerative dermatitis as used in literature is somehow confusing because on the one hand this skin disorder is associated with bacterial growth and on the other hand it is a synonym for a chronic sporadic disease of adult sows with unknown aetiology. Thus, we propose the terminus ‘Porcine Ulcerative Dermatitis Syndrome (PUDS)’ for the latter to distinguish between these two disease complexes. This syndrome could be identified by clinical and pathological examinations in six sows, that were submitted to the clinic. Epidermal ulcers could be found nearly all over the body, but teats were always spared. Haematological examination showed a slight anaemia but physiological leucocyte counts. However, lymphopenia (x = 44.8%), granulocytosis (x = 42.0%) and an increased number of monocytes (x = 13.1%) could be found. Histologically, a lymphoplasmacytic and granulohistiocytic infiltration in the corium was most prominent. In some cases, a moderate leucocytoclastic vasculitis and perivasculitis could be seen at the dermo‐epidermal border. Additionally, a multifocal interstitial nephritis with lymphoplasmacytic infiltration was a prominent feature in all animals. Participation of an immune complex associated disorder can be assumed when regarding histological findings as skin lesions in combination with glomerulonephritis are a common feature of such diseases. Also, IgG levels were elevated two‐ to fourfold in all affected sows when compared with healthy control pigs. This supports the hypothesis that not only T cells, as shown previously, but also the humoral branch of the immune system is involved in the aetiology of PUDS.