bluuebunny/crossref_metadata_2025 huggingface dataset:
Trauma‐Induced Cutaneous Pigmentation from Tetracycline: A Case Report
Authors: Hawfield Wesley , Goodrich Robert , Warren Simon , Morrell Dean
Published: April 2004

Abstract:  Tetracycline is well known to cause pigmentation of various body tissues, such as teeth, bone, lunulae, conjunctivae, and lymph nodes. It has not been reported to cause discoloration of the skin, unlike its semisynthetic derivative, minocycline. We report an 18‐year‐old girl who developed lower extremity pigmentation in areas of trauma during treatment with tetracycline.
Salt stress‐induced chloride flux: a study using transgenic Arabidopsis expressing a fluorescent anion probe
Authors: Lorenzen Inken , Aberle Thomas , Plieth Christoph
Published: April 2004

Salt stress leads to massive accumulation of toxic levels of Na+ and Cl− ions in plants. By using the recombinant fluorescent probe CLOMELEON, we demonstrate passive anion flux under salt stress. Chloride influx is restricted in the presence of divalent cations like Mg2+ and Ca2+, and completely blocked by La3+. The amount but not the rate of the reported chloride uptake is independent from the kind of corresponding permeable cation (K+ versus Na+), external pH and magnitude of osmotic stress. Cl− efflux however seems to involve stretch‐activated transport. From the influence of Ca2+ on reported changes of cytosolic anion concentrations, we speculate that transport mechanisms of Cl− and Na+ might be thermodynamically coupled under saline conditions.
Identification of Botrytis cinerea susceptibility loci in Arabidopsis thaliana
Authors: Denby Katherine J. , Kumar Pavan , Kliebenstein Daniel J.
Published: April 2004

Botrytis cinerea is a major pathogen of fruit and vegetable crops causing both pre‐ and post‐harvest grey mould. We have analysed 16 Arabidopsis thaliana ecotypes for natural variation in B. cinerea susceptibility. Susceptibility was associated with lower camalexin accumulation, and three ecotypes (Cape Verdi Islands (Cvi‐0), Slavice (Sav‐0) and Kindalville (Kin‐0)) showed differential susceptibility to the two B. cinerea isolates used. Subsequently, to better understand the genetic control of grey mould disease, we assayed the Arabidopsis Landsberg erecta (Ler) × Columbia (Col‐0) recombinant inbred population with the two isolates, and identified multiple small‐to‐medium‐effect quantitative trait loci (QTL) governing susceptibility. Interestingly, the QTL for each isolate are distinct, suggesting that different mechanisms govern defence against these two isolates. Two QTL for each isolate exhibited epistatic interactions with specific allele combinations generating heightened B. cinerea susceptibility.
Expression of rolB in tobacco flowers affects the coordinated processes of anther dehiscence and style elongation
Authors: Cecchetti Valentina , Pomponi Mirella , Altamura Maria Maddalena , Pezzotti Mario , Marsilio Sonia , D’Angeli Simone , Tornielli Giovanni Battista , Costantino Paolo , Cardarelli Maura
Published: April 2004

The effect of auxin on stamen and pistil development in tobacco flowers was investigated by means of the localized expression of rolB (root loci B), an Agrobacterium oncogene that increases auxin sensitivity in a cell‐autonomous fashion. When rolB is driven by the promoter of the meiosis‐specific Arabidopsis gene DMC1 (disrupted meiotic cDNA 1), expression occurs earlier in male than in female developing organs, resulting in a delay in anther dehiscence with respect to normal timing of pistil development. As a consequence of this developmental uncoupling, self‐pollination is prevented in pDMC1:rolB plants. Histological analysis of pDMC1:GFP plants indicates that in tobacco, this promoter is active not only in meiocytes but also in somatic tissues of the anther. In contrast, simultaneous expression of rolB in anther and pistil somatic tissues, achieved by expressing a construct containing rolB under the control of the promoter of the petunia gene FBP7 (floral binding protein 7), results in a concomitant delay of both anther dehiscence and pistil development without affecting self‐pollination of the plants. Analysis of plants harboring the pFBP7:GUS construct shows that in tobacco, this promoter is active not only in the ovules, as described for petunia, but also in pistil and anther somatic tissues involved in the dehiscence program. The delay in anther dehiscence and pistil development could be phenocopied by exogenous application of auxin. Jasmonic acid (JA) could not rescue the delay in anther dehiscence. These results suggest that auxin plays a key role in the timing of anther dehiscence, the dehiscence program is controlled by the somatic tissues of the anther, and auxin also regulates pistil development.
A novel polymorphism in the cytoplasmic region of the human immunoglobulin A Fc receptor gene*
Authors: Jasek M. , Mańczak M. , Sawaryn A. , Obojski A. , Wiśniewski A. , Łuszczek W. , Kuśnierczyk P.
Published: April 2004

The Fc receptor for immunoglobulin A (IgA), FcαRI, is expressed on several types of myeloid cells, and activates them upon ligand binding. However, binding of IgA to the extracellular domain of the receptor requires previous stimulation of the cell by cytokines, and the cytoplasmic tail of FcαRI has been shown to play a role in this. Therefore, polymorphism in this region might affect this process. However, no changes in the amino acid sequence in this region of the FcαRI have so far been reported. Here, we describe for the first time a single nucleotide polymorphism in exon 5 of the immunoglobulin A Fc receptor (FCAR) gene leading to a Ser→Gly substitution at position 248 of the mature FcαRI protein. Prediction of structural features suggests some changes that may affect the function of the protein to some extent. However, the Gly248 variant is quite common (4% homozygotes and 38% heterozygotes) in healthy population, suggesting a weak effect, if any, on function, at least in heterozygotes.
MICA polymorphism in a sample of the São Paulo population, Brazil
Authors: Marin M. L. C. , Savioli C. R. , Yamamoto J. H. , Kalil Jorge , Goldberg A. C.
Published: April 2004

The major histocompatibility complex (MHC) class I chain‐related A (MICA) gene, located near HLA‐B, codes for protein products with structural similarities to those of classical MHC class I genes, but which neither bind β2‐microglobulin nor present peptide. Expressed predominantly on gastrointestinal and tumour epithelial cells, they are stress‐induced and interact with C‐type lectin like receptor (NKG2D) on γδ, αβ CD8+ T cells and natural killer (NK) cells. MICA is highly polymorphic, with 54 extracellular allelic sequences described. We typed 200 healthy subjects in a sample of the São Paulo population by extended polymerase chain reaction–sequence‐specific primers (PCR‐SSP) to characterize the MICA polymorphism and analysed MICA/HLA‐B linkage disequilibrium. The MICA*008 group (g) was predominant (47%), with several HLA‐B associations. Rare combinations MICA*008g‐HLA‐B37, MICA*008g‐B72 and MICA*010‐HLA‐B52 were detected. Given the extent of this polymorphism and its possible relevance for disease association, we determined MICA and HLA‐B alleles in 33 Behçet's patients, in an attempt to clarify the associated genetic marker. Our results showed an increase of MICA*006, but not MICA*009, in the patient group (6/33) compared with controls (3/200) (18.2% vs. 1.5%; Pc = 0.005). Both alleles were always in association with HLA‐B51, suggesting that HLA‐B is indeed the primary susceptibility locus (P = 0.00008) and that MICA*006 may be an additional risk factor.
Lack of association of human chemokine receptor gene polymorphisms CCR2‐64I and CCR5‐Δ32 with autoimmune Addison's disease
Authors: Gambelunghe G. , Ghaderi M. , Gharizadeh B. , Brozzetti A. , Tortoioli C. , Del Sindaco P. , Sanjeevi C. B. , Hjelmström P. , Sirsjö A. , Nyren P. , Santeusanio F. , Falorni A.
Published: April 2004

The attraction of leukocytes to tissues is essential for inflammation and the initiation of the autoimmune reaction. The process is controlled by chemokines, which are chemotactic cytokines. We investigated whether human chemokine receptor gene polymorphisms, namely CCR5‐Δ32 and CCR2‐64I, are associated with susceptibility to autoimmune Addison's disease. Genotyping was performed in 56 patients and 127 healthy controls by a new method using pyrosequencing for CCR2‐64I and by polymerase chain reaction and detecting gel for CCR5‐Δ32. None of the CCR2 or CCR5 alleles was found to be associated, either positively or negatively, with disease risk. Our results indicate that the CCR2‐64I and CCR5‐Δ32 gene polymorphisms do not play a major role in conferring genetic risk for, and/or protection against, autoimmune Addison's disease.
T‐cell receptor BV gene segment polymorphisms in healthy elderly individuals from the south of Brazil
Authors: Peres A. , Hünemeier T. , Da Cruz I. M. , Nardi N. B. , Chies J. A. B.
Published: April 2004

The ageing of the immune system (immunosenescence) is believed to be involved in both morbidity and mortality in elderly humans due to a higher incidence of infections, autoimmune diseases, cancers and other pathological situations. As any specific immune response involves recognition of antigens by T cells, the ability to develop a given immune response is also dependent on the T‐cell repertoire available at a given time point. Different T‐cell receptor beta variable segment (BV) (TCRBV) gene segment alleles have been associated with diseases in various human populations. In the present work we analysed the allelic frequencies of four biallelic polymorphisms in TCRBV gene segments (TCRBV3S1, TCRBV13S5, TCRBV13S6 and TCRBV18) in healthy elderly human subjects (80 years old or more) from the south of Brazil, where life expectancies reach similar levels to those observed in developed countries. Except for allele 2 of the TCRBV13S6 polymorphism, which was more frequent in elderly than in young individuals (P = 0.0105), there were no differences in allele or genotype frequencies between young and elderly individuals. The data suggest that there is no direct correlation between the TCRBV3S1, TCRBV13S5 and TCRBV18 polymorphisms analysed and healthy senescence in this particular group of elderly individuals. The higher frequency of TCRBV13S6 allele 2 in healthy elderly individuals should be confirmed in other samples to establish the significance of this finding.
No significant association between mutations in exons 6 and 8 of the autoimmune regulator (AIRE) gene and inflammatory bowel disease
Authors: Török H.‐P. , Tonenchi L. , Glas J. , Schiemann U. , Folwaczny C.
Published: April 2004

Various autoantibodies have been described in patients with inflammatory bowel disease. The autoimmune regulator (AIRE) functions as a transcription factor in cells responsible for the induction and maintenance of immunological tolerance. In contrast to classic autoimmune disorders, polymorphisms of the AIRE gene are not associated with inflammatory bowel disease, despite the presence of disease‐specific autoantibodies.
Provision of Epstein‐Barr virus‐transformed B‐cell lines in a routine tissue typing laboratory: practicalities and applications
Authors: Bass H. , Walters R. , Darke C.
Published: April 2004

A bank of Epstein‐Barr virus (EBV)‐transformed B‐lymphoblastoid cell lines (BCLs) has been established in this laboratory over the last 7 years. Novel, rare and unusual HLA phenotypes were highlighted during routine clinical testing and during typing of volunteer haematopoietic stem cell donors for the Welsh Bone Marrow Donor Registry. The BCLs are routinely used as laboratory reagents, as a source of DNA for reference purposes and for research work. Consenting donors are tested for hepatitis B surface antigens (HBsAg), human immunodeficiency virus (HIV) and hepatitis C virus (HCV) before their B‐lymphocytes are transformed. Strict bio‐identity checks are performed before and after transformation and after each BCL expansion. Each BCL is tested regularly for mycoplasma contamination. A total of 230 blood samples were transformed. One hundred and fifty‐nine sterile samples produced 157 BCLs (98.7% success), while 71 non‐sterile samples produced 50 BCLs (70.4% success), giving an overall success rate of 90.0%. Fifteen of the transformation failures have since been repeated successfully. Factors contributing to the high success rate and reasons for the 23 failed transformations are discussed. The successful development and use of pools of BCLs for HLA antibody screening by flow cytometry are described. Whilst certain training and health and safety issues require close attention, it is clear that EBV transformation of B lymphocytes and/or the use of BCLs is feasible in a routine tissue typing laboratory and that BCLs are a valuable resource. Careful adherence to the methods and procedures detailed here should virtually guarantee successful transformation (98.7%) from good quality, sterile whole blood samples.
The MICA‐A4 triplet repeats polymorphism in the transmembrane region confers additional risk for development of psoriatic arthritis in the Croatian population
Authors: Grubić Z. , Perić P. , Èeèuk‐Jeličić E. , Žunec R. , Štingl K. , Ćurković B. , Kerhin‐Brkljačić V.
Published: April 2004

The aim of this study was to investigate possible differences in the frequencies of alleles at the HLA loci and at microsatellite loci within the HLA region among patients suffering from psoriatic arthritis (PsA) and healthy controls. Fifty‐eight Croatian PsA patients (28 male and 30 female) and 157 healthy unrelated controls were typed for HLA alleles (A, B, Cw and DRB1) by the polymerase chain reaction–sequence‐specific primers (PCR‐SSP) method, while microsatellite alleles (D6S265, D6S273, MHC class I chain‐related gene (MICA) and MIB) were analysed by electrophoresis in an ALFexpress sequencer (Pharmacia Biotech, Uppsala, Sweden). The findings from this study were: (1) the frequencies of B*39 and B*57 were significantly increased in PsA patients; (2) differences in the frequencies of B*13 and B*27 were not statistically significant after correction; (3) the B*0702, B*18, and B*38 alleles were decreased in patients only before correction; (4) none of the alleles at other HLA loci tested were associated with PsA in Croatia; (5) polymorphism at D6S265, D6S273, and MIB microsatellites in patients did not show any statistically significant differences when compared to controls; (6) the increase in the MICA‐A4 allele frequency in PsA patients was independent of the B*39 and B*57 alleles.
Effective and safe use of recombinant factor VIIa (NovoSeven®) in elderly mild haemophilia A patients with high‐titre antibodies against factor VIII
Authors: Leebeek F. W. G. , Kappers‐Klunne M. C. , Jie K. S. G.
Published: April 2004

Summary.  Three patients with mild haemophilia A who developed high‐titre antibodies against factor VIII at high age are reported. These patients had only a limited number of exposure days of FVIII concentrates in the past. The patients had to undergo surgery or presented with recurrent bleeding episodes. Treatment with recombinant FVIIa (rFVIIa) was effective and safe. Despite the high age and the presence of coronary heart disease in one of the patients, no adverse events or thrombotic complications occurred. These cases illustrate that the physician should always be alert on the development of inhibitors, also in elderly patients with mild haemophilia, in whom FVIII inhibitors had never been detectable before and that treatment with rFVIIa was effective and well‐tolerated.
Cryoprecipitate‐removed plasma ‘cryo‐removed plasma’ as a source of factor IX in the treatment of haemophilia B
Authors: Lepatan L. M. P. , Hernandez F. G. , Montoya M. M. , Ong J. B. , Rodriguez E. V. , Chua M. N.
Published: April 2004

Summary.  Fresh frozen plasma (FFP) is the main source of factor IX (FIX) in the treatment of bleeding episodes of haemophilia B in the Philippines. Cryoprecipitate‐removed plasma otherwise known in the Philippines as cryosupernate, is a by‐product of cryoprecipitate preparation. These blood products expire in storage or are just thrown‐ away because of less demand for clinical use. By theory, this product should have almost the same amount of FIX as in FFP, therefore can be used in the treatment of haemophilia B. There is no local data on the actual FIX content of the cryoprecipitate‐removed plasma. Hence, the authors established these data to support the use of this product. Eighty‐three bags of cryoprecipitate‐removed plasma received from three different blood banks in Manila, Philippines were tested for FIX activity using an activated partial thromboplastin time (APTT)‐based one‐stage FIX assay. The FIX content in each bag of cryoprecipitate‐removed plasma was calculated by multiplying its volume in mL with that of FIX activity per mL of plasma measured in vitro. The total mean FIX content per bag was 212.20 U (±88.98) exceeding the contents set by the American Association of Blood Banks (AABB, 70–90 U). The mean FIX activity per bag was 127.62% (±38.23) with the mean volume of 164.28 mL (±52.23). Statistically significant difference on volume (P = 0.000) was found across the three sources resulting to a significant variation of the actual FIX content (P = 0.000).
Management of von Willebrand disease: a guideline from the UK Haemophilia Centre Doctors' Organization
Authors: Pasi K. J. , Collins P. W. , Keeling D. M. , Brown S. A. , Cumming A. M. , Dolan G. C. , Hay C. R. M. , Hill F. G. H. , Laffan M. , Peake I. R.
Published: April 2004

Summary.  von Willebrand disease (VWD) is the commonest inherited bleeding disorder. The aim of therapy for VWD is to correct the two defects of haemostasis in this disorder, impaired primary haemostasis because of defective platelet adhesion and aggregation and impaired coagulation as a result of low levels of factor VIII. The objective of this guideline is to inform individuals making choices about the treatment and management of VWD including the use of therapeutic products. This is the second edition of this UK Haemophilia Centre Doctors' Organization (UKHCDO) guideline and supersedes the previous edition which was published in 1994.
Haemorrhagic symptoms in patients with combined factors V and VIII deficiency in north‐eastern Iran
Authors: Mansouritorgabeh H. , Rezaieyazdi Z. , Pourfathollah A. A. , Rezai J. , Esamaili H.
Published: April 2004

Summary.  Of the six types of dual coagulation factors deficiency, combined factors V and VIII are the most common type, a few cases of this disease have been reported in different populations. This accounts for the relatively low number of cases reported so far. Our report, which included 19 patients, is the second largest group that has been reported from one centre in north‐eastern Iran. The most frequent spontaneous bleeding symptoms were epistaxis and haemarthrosis, and the most frequent traumatic bleeding symptoms were bleeding after dental extraction and bleeding after cutting any part of the body. It seemed that dual coagulation FV and FVIII deficiency is as severe as single coagulation factor (V or VIII) deficiency.
A comparative in vitro evaluation of six von Willebrand factor concentrates
Authors: Lethagen S. , Carlson M. , Hillarp A.
Published: April 2004

Summary.  The efficacy of von Willebrand factor (VWF) concentrates for treatment of von Willebrand disease (VWD) is dependent on their content of VWF and factor VIII (FVIII).

Study objectives:  To measure the content and quality of VWF and FVIII in six VWF concentrates: Haemate‐P (Aventis Behring), Immunate (Baxter Bioscience), Koate (Bayer Corp.), 8Y (BPL), Innobrand (LFB) and Facteur Willebrand (LFB).

Methods:  The VWF antigen content (VWF:Ag), ristocetin cofactor activity (VWF:RCo), collagen‐binding activity (VWF:CB), VWF multimers with electrophoresis and densitometry, FVIII activity and total protein content.

Results:  Specific activity (VWF:RCo/total protein) varied considerably (4.7–129.5 IU mg−1). Activity measures, VWF:RCo and VWF:CB, correlated well, but we found no correlation between any of these and VWF:Ag. The content of high‐molecular weight multimer (HMWM) was normal or close to normal in Haemate‐P, Innobrand and Facteur Willebrand, moderately reduced in Koate and 8Y, and significantly reduced in Immunate. The HMWM content correlated significantly with the VWF:RCo/VWF:Ag ratio. Only Haemate‐P, Innobrand and Facteur Willebrand had VWF:RCo/VWF:Ag ratios >0.7. We found large differences in the content of FVIII and in the FVIII/VWF:RCo ratio. Facteur Willebrand had the lowest (0.02) and Immunate the highest (6.00) ratio.

Conclusion:  Treating physicians must be aware of the large differences between different VWF concentrates and the potential clinical implications. Concentrates lacking HMWM are probably less efficient for mucosal bleedings. FVIII is most important for surgical bleedings, but concentrates with high FVIII/VWF‐ratio may induce very high FVIII levels with increased risk of thrombosis. A low FVIII content may be preferable except in case of acute surgery.
The diagnosis of von Willebrand disease: a guideline from the UK Haemophilia Centre Doctors’ Organization
Authors: Laffan M. , Brown S. A. , Collins P. W. , Cumming A. M. , Hill F. G. H. , Keeling D. , Peake I. R. , Pasi K. J.
Published: April 2004

Summary.  von Willebrand disease (VWD) is the commonest inherited bleeding disorder. However, despite an increasing understanding of the pathophysiology of VWD, the diagnosis of VWD is frequently difficult because of uncertainty regarding the relationship between laboratory assays and function in vivo. The objective of this guideline is to provide contemporary advice on a rational approach to the diagnosis of VWD. This is the second edition of this UK Haemophilia Centre Doctors’ Organisation (UKHCDO) guideline and supersedes the previous edition which was published in 1997.
Molecular pathology of haemophilia B: identification of five novel mutations including a LINE 1 insertion in Indian patients
Authors: Mukherjee S. , Mukhopadhyay A. , Banerjee D. , Chandak G. R. , Ray K.
Published: April 2004

Summary.  Heterogeneous mutations in factor IX (FIX) gene cause haemophilia B and a large number of mutations have been characterized. However, reports on gene defects among Indian haemophilia B patients are rare despite a high estimate of such patients in the country. We report identification of 22 independent mutations including five novel mutations in 24 unrelated patients. The novel gene defects include two point mutations, two deletions and one insertion of a LINE 1 element. Majority of the mutations (14 of 24) occurred on the same haplotype background, but do not suggest any founder effect. Direct identification of mutations can be utilized to perform the carrier detection and prenatal diagnosis, especially in families with isolated patients.
Identification of three F5 gene mutations associated with inherited coagulation factor V deficiency in two Chinese pedigrees
Authors: Fu Q.‐H. , Zhou R.‐F. , Liu L.‐G. , Wang W.‐B. , Wu W.‐M. , Ding Q.‐L. , Hu Y.‐Q. , Wang X.‐F. , Wang Z.‐Y. , Wang H.‐L.
Published: April 2004

Summary.  To investigate the molecular defects in two Chinese pedigrees with inherited factor V (FV) deficiency.

A 37‐year‐old male (proband 1) and an 18‐month‐old boy (proband 2) were diagnosed as inherited coagulation FV deficiency by severely reduced plasma levels of FV activity and antigen. All 25 exons and their flanking sequence of F5 gene were amplified by polymerase chain reaction (PCR) for both probands and the PCR products were directly sequenced. Total RNA was extracted from the peripheral lymphocytes of proband 1 for detecting the changes at mRNA level.

The homozygous deletion IVS8 −2A>G was identified in the F5 gene of proband 1 and complementary DNA (cDNA) analysis revealed the abolishment of the canonical splicing site by the mutation and the activation of the cryptic acceptor site 24 bp upstream instead. The insertion introduced eight additional amino acids (AA) into the FV protein. Two heterozygous mutations of F5 gene were discovered in proband 2. The 2238‐9del AG in exon 13 introduced a premature termination code at 689 AA and the substitution of G6410 by T in exon 23 lead to the missense mutation Gly2079Val.

Three F5 gene mutations, IVS8 −2A>G, 2238‐9del AG and G6410T, have been identified in two Chinese pedigree with congenital FV deficiency, respectively.
Experimental haemophilic synovitis: rationale and development of a murine model of human factor VIII deficiency
Authors: Valentino L. A. , Hakobyan N. , Kazarian T. , Jabbar K. J. , Jabbar A. A.
Published: April 2004

Summary.  Haemophilia is a genetic disease as a result of the deficiency of blood coagulation factor VIII or IX. Bleeding is common, especially into joints where an inflammatory, proliferative synovitis develops resulting in a debilitating arthritis, haemophilic arthropathy. The pathogenesis of blood‐induced haemophilic synovitis (HS) is poorly understood. The gross, microscopic and ultrastructural changes that occur in the synovial membrane following human and experimental hemarthrosis have been described. Repeated episodes of bleeding induce synoviocyte hypertrophy and hyperplasia, an intense neovascular response and inflammation of the synovial membrane. The component(s) in blood that initiates these changes is(are) not known, although iron is often proposed as one possibility. Here, we describe a novel murine model of human haemophilia A, which facilitates the examination of large number of animals and tissue specimens. The effects of hemarthrosis on the physical, gross and microscopic changes evoked following joint bleeding are described. Controlled, blunt trauma to the knee joint consistently resulted in joint swelling because of a combination of bleeding and inflammation. Hemosiderin was found in the synovial membrane. Similar to hemarthrosis in human haemophilia, joint bleeding resulted in acute morbidity evidenced by inactivity, weight loss and immobility. With time the animals recovered. The model of experimental murine HS described here has utility in the study of the pathogenesis of HS. This is the first of a series of articles, which will discuss the pathophysiology and characterize the model, with comparison of his model to others which have been published previously. It should provide a useful model to test potential therapeutic interventions.
Salicylic acid dependent signaling promotes basal thermotolerance but is not essential for acquired thermotolerance in Arabidopsis thaliana
Authors: Clarke Shannon M. , Mur Luis A. J. , Wood Jacqueline E. , Scott Ian M.
Published: April 2004

Salicylic acid (SA) is reported to protect plants from heat shock (HS), but insufficient is known about its role in thermotolerance or how this relates to SA signaling in pathogen resistance. We tested thermotolerance and expression of pathogenesis‐related (PR) and HS proteins (HSPs) in Arabidopsis thaliana genotypes with modified SA signaling: plants with the SA hydroxylase NahG transgene, the nonexpresser of PR proteins (npr1) mutant, and the constitutive expressers of PR proteins (cpr1 and cpr5) mutants. At all growth stages from seeds to 3‐week‐old plants, we found evidence for SA‐dependent signaling in basal thermotolerance (i.e. tolerance of HS without prior heat acclimation). Endogenous SA correlated with basal thermotolerance, with the SA‐deficient NahG and SA‐accumulating cpr5 genotypes having lowest and highest thermotolerance, respectively. SA promoted thermotolerance during the HS itself and subsequent recovery. Recovery from HS apparently involved an NPR1‐dependent pathway but thermotolerance during HS did not. SA reduced electrolyte leakage, indicating that it induced membrane thermoprotection. PR‐1 and Hsp17.6 were induced by SA or HS, indicating common factors in pathogen and HS responses. SA‐induced Hsp17.6 expression had a different dose–response to PR‐1 expression. HS‐induced Hsp17.6 protein appeared more slowly in NahG. However, SA only partially induced HSPs. Hsp17.6 induction by HS was more substantial than by SA, and we found no SA effect on Hsp101 expression. All genotypes, including NahG and npr1, were capable of expression of HSPs and acquisition of HS tolerance by prior heat acclimation. Although SA promotes basal thermotolerance, it is not essential for acquired thermotolerance.
ABA activates ADPR cyclase and cADPR induces a subset of ABA‐responsive genes in Arabidopsis
Authors: Sánchez Juan‐Pablo , Duque Paula , Chua Nam‐Hai
Published: April 2004

Cyclic ADP‐ribose (cADPR) was previously shown to activate transient expression of two abscisic acid (ABA)‐responsive genes in tomato cells. Here, we show that the activity of the enzyme responsible for cADPR synthesis, ADP‐ribosyl (ADPR) cyclase, is rapidly induced by ABA in both wild‐type (WT) and abi1‐1 mutant Arabidopsis plants in the absence of protein synthesis. Furthermore, in transgenic Arabidopsis plants, induced expression of the Aplysia ADPR cyclase gene resulted in an increase in ADPR cyclase activity and cADPR levels, as well as elevated expression of ABA‐responsive genes KIN2, RD22, RD29a, and COR47 (although to a lesser extent than after ABA induction). Genome‐wide profiling indicated that about 28% of all ABA‐responsive genes in Arabidopsis are similarly up‐ and downregulated by cADPR and contributed to the identification of new ABA‐responsive genes. Our results suggest that activation of ADPR cyclase is an early ABA‐signaling event partially insensitive to the abi1‐1 mutation and that an increase in cADPR plays an important role in downstream molecular and physiological ABA responses.
New in silico insight into the synteny between rice (Oryza sativa L.) and maize (Zea mays L.) highlights reshuffling and identifies new duplications in the rice genome
Authors: Salse Jérôme , Piégu Benoit , Cooke Richard , Delseny Michel
Published: April 2004

A unigene set of 1411 contigs was constructed from 2629 redundant maize expressed sequence tags (ESTs) mapped on the maizeDB genetic map. Rice orthologous sequences were identified by blast alignment against the rice genomic sequence. A total of 1046 (74%) maize contigs were associated with their corresponding homologues in the rice genome and 656 (47%) defined as potential orthologous relationships. One hundred and seventeen (8%) maize EST contigs mapped to two distinct loci on the maize genetic map, reflecting the tetraploid nature of the maize genome. Among 492 mono‐locus contigs, 344 (484 redundant ESTs) identify collinear blocks between maize chromosomes 2 and 4 and a single rice chromosome, defining six new collinear regions. Fine‐scale analysis of collinearity between rice chromosomes 1 and 5 with maize chromosomes 3, 6 and 8 shows the presence of internal rearrangements within collinear regions. Mapping of maize contigs to two distinct loci on the rice sequence identifies five new duplication events in rice. Detailed analysis of a duplication between rice chromosomes 1 and 5 shows that 11% of the annotated genes from the chromosome 1 locus are found duplicated on the chromosome 5 paralogous counterpart, indicating a high degree of re‐organisations. The implications of these findings for map‐based cloning in collinear regions are discussed.
Development and evaluation of an Arabidopsis whole genome Affymetrix probe array
Authors: Redman Julia C. , Haas Brian J. , Tanimoto Gene , Town Christopher D.
Published: April 2004

We describe the development of a high‐density Arabidopsis‘whole genome’ oligonucleotide probe array for expression analysis (the Affymetrix ATH1 GeneChip® probe array) that contains approximately 22 750 probe sets. Precedence on the array was given to genes for which either expression evidence or a credible database match existed. The remaining space was filled with ‘hypothetical’ genes. The new ATH1 array represents approximately 23 750 genes of which 60% were detected in RNA from cultured seedlings. Sensitivity of the array, determined using spiking controls, was approximately one transcript per cell. The array demonstrated high technical reproducibility and concordance with real‐time PCR results. Indole‐3 acetic acid (IAA)‐induced changes in gene expression were used for biological validation of the array. A total of 222 genes were significantly upregulated and 103 significantly downregulated by exposure to IAA. Of the genes whose products could be functionally classified, the largest specific classes of upregulated genes were transcriptional regulators and protein kinases, many fewer of which were represented among the downregulated genes. Over one‐third of the auxin‐regulated genes have no known function, although many belong to gene families with members that have previously been shown to be auxin regulated. For the 6714 genes represented both on this and the earlier Arabidopsis Genome (AG) array, both signal intensities and gene expression ratios were very similar. Mapping of the oligonucleotides on the ATH1 array to the latest (version 4.0) annotation showed that over 95% of the probe sets (based on version 2.0 annotation) still fully represented their original target genes.
A novel calmodulin‐binding protein functions as a negative regulator of osmotic stress tolerance in Arabidopsis thaliana seedlings
Authors: Perruc Elian , Charpenteau Martine , Ramirez Bertha Cecilia , Jauneau Alain , Galaud Jean‐Philippe , Ranjeva Raoul , Ranty Benoît
Published: April 2004

A clone for a novel Arabidopsisthaliana calmodulin (CaM)‐binding protein of 25 kDa (AtCaMBP25) has been isolated by using a radiolabelled CaM probe to screen a cDNA expression library derived from A. thaliana cell suspension cultures challenged with osmotic stress. The deduced amino acid sequence of AtCaMBP25 contains putative nuclear localization sequences and shares significant degree of similarity with hypothetical plant proteins only. Fusion of the AtCaMBP25 coding sequence to reporter genes targets the hybrid protein to the nucleus. Bacterially expressed AtCaMBP25 binds, in a calcium‐dependent manner, to a canonical CaM but not to a less conserved isoform of the calcium sensor. AtCaMBP25 is encoded by a single‐copy gene, whose expression is induced in Arabidopsis seedlings exposed to dehydration, low temperature or high salinity. Transgenic plants overexpressing AtCaMBP25 exhibits an increased sensitivity to both ionic (NaCl) and non‐ionic (mannitol) osmotic stress during seed germination and seedling growth. By contrast, transgenic lines expressing antisense AtCaMBP25 are significantly more tolerant to mannitol and NaCl stresses than the wild type. Thus, the AtCaMBP25 gene functions as a negative effector of osmotic stress tolerance and likely participates in stress signal transduction pathways.
An Arabidopsis dynamin‐related protein, DRP3A, controls both peroxisomal and mitochondrial division
Authors: Mano Shoji , Nakamori Chihiro , Kondo Maki , Hayashi Makoto , Nishimura Mikio
Published: April 2004

Peroxisomes undergo dramatic changes in size, shape, number, and position within the cell, but the division process of peroxisomes has not been characterized. We screened a number of Arabidopsis mutants with aberrant peroxisome morphology (apm mutants). In one of these mutants, apm1, the peroxisomes are long and reduced in number, apparently as a result of inhibition of division. We showed that APM1 encodes dynamin‐related protein 3A (DRP3A), and that mutations in APM1/DRP3A also caused aberrant morphology of mitochondria. The transient expression analysis showed that DRP3A is associated with the cytosolic side of peroxisomes. These findings indicate that the same dynamin molecule is involved in peroxisomal and mitochondrial division in higher plants. We also report that the growth of Arabidopsis, which requires the cooperation of various organelles, including peroxisomes and mitochondria, is repressed in apm1, indicating that the changes of morphology of peroxisomes and mitochondria reduce the efficiency of metabolism in these organelles.
DISTORTED2 encodes an ARPC2 subunit of the putative Arabidopsis ARP2/3 complex
Authors: El‐Din El‐Assal Salah , Le Jie , Basu Dipanwita , Mallery Eileen L. , Szymanski Daniel B.
Published: April 2004

Arabidopsis trichomes are unicellular, branched structures that have highly constrained requirements for the cytoskeleton. The ‘distorted group’ genes function downstream from microtubule‐based branch initiation, and are required during the actin‐dependent phase of polarized stalk and branch expansion. Of the eight known ‘distorted group’ genes, a subset encode homologs of ARP2/3 complex subunits. In eukaryotic cells, the seven‐protein ARP2/3 complex nucleates actin filament networks that push on the plasma membrane and organelles. In plants cells, the existence and function of an ARP2/3 complex is unclear. In this paper, we report that DISTORTED2 (DIS2) encodes a paralogue of the ARP2/3 complex subunit ARPC2. DIS2 has ARPC2 activity, based on its ability to rescue the growth defects of arpc2 (arc35Δ) null yeast cells. Like known ARPC2s, DIS2 physically interacts with ARPC4. Mutations in DIS2 cause a distorted trichome phenotype, defects in cell–cell adhesion, and a modest reduction in shoot FW. The actin cytoskeleton in dis2 trichomes is extensive, but developing branches fail to generate and maintain highly organized cytoplasmic actin bundles.
Induction of ASCORBATE PEROXIDASE 2 expression in wounded Arabidopsis leaves does not involve known wound‐signalling pathways but is associated with changes in photosynthesis
Authors: Chang Christine Chi‐Chen , Ball Louise , Fryer Michael J. , Baker Neil R. , Karpinski Stanislaw , Mullineaux Philip M.
Published: April 2004

ASCORBATE PEROXIDASE 2 (APX2) encodes a key enzyme of the antioxidant network. In excess light‐stressed Arabidopsis leaves, photosynthetic electron transport (PET), hydrogen peroxide (H2O2) and abscisic acid (ABA) regulate APX2 expression. Wounded leaves showed low induction of APX2 expression, and when exposed to excess light, APX2 expression was increased synergistically. Signalling pathways dependent upon jasmonic acid (JA), chitosan and ABA were not involved in the wound‐induced expression of APX2, but were shown to require PET and were preceded by a depressed rate of CO2 fixation. This led to an accumulation of H2O2 in veinal tissue. Diphenyl iodonium (DPI), which has been shown previously to be a potent inhibitor of H2O2 accumulation in the veins of wounded leaves, prevented induction of APX2 expression probably by inhibition of PET. Thus, the weak induction of APX2 expression in wounded leaves may require H2O2 and PET only. As in other environmental stresses, wounding of leaves resulted in decreased photosynthesis leading to increased reactive oxygen species (ROS) production. This may signal the induction of many ‘wound‐responsive’ genes not regulated by JA‐dependent or other known JA‐independent pathways.
The Lumbosacral Junction in Working German Shepherd Dogs – Neurological and Radiological Evaluation
Authors: Scharf G. , Steffen F. , Grünenfelder F. , Morgan J. P. , Flückiger M.
Published: April 2004

The clinical and radiological incidence of lumbosacral (LS) disease was studied on 57 German Shepherd dogs (GSDs) used in active service. The study included a clinical history, a neurological examination, and plain radiographs of the caudal lumbar vertebrae. The neurological examinations revealed lower back pain and/or neural deficits in 21 dogs, of which 14 had a history of pain or pelvic gait abnormalities. Radiographic findings were spondylosis at L7–S1, degeneration of L7–S1 disc, LS malalignment, transitional LS vertebrae and/or primary spinal canal stenosis in 15 dogs with neurological abnormalities and/or back pain and in 18 dogs with no clinical signs. No correlation between the neurological and the radiographic findings were found. This study demonstrates that even prominent radiographic LS abnormalities are of minimal value in the evaluation of LS disease in the GSD.
Effect of a Dietary n‐6 Polyunsaturated Fatty Acid Supplement on Distinct Immune Functions of Goats
Authors: Thanasak J. , Rutten V. P. M. G. , Schonewille J. Th. , Hoek A. , Beynen A. C. , Noordhuizen J. P. T. M. , Müller K. E.
Published: April 2004

The aim of this study was to investigate whether dietary n‐6 polyunsaturated fatty acids (PUFA) alter immune functions as lymphocyte blastogenesis, expression of lymphocyte and monocyte markers, and cell adhesion molecules of the integrin family in goats. Dutch White castrated male goats received a diet either rich in olive oil (control group, n = 4) or rich in corn oil (test group, n = 4) over a period of 3 weeks. The animals in the test group had significantly higher linoleic acid (LA, 18:2n‐6) levels in plasma on days 7, 14 and 21 of the diet than control animals. This was also true for washed erythrocyte membranes at day 21. A significant reduction in the percentage of α‐4 integrin (CD49d) expressing lymphocytes was observed in goats fed LA rich corn oil at day 21. However, no changes were observed in either lymphocyte proliferative responses to ConA and PHA or the expression of other lymphocyte/monocyte markers. In conclusion, feeding corn oil rich in LA caused significant increases in the proportion of LA in the fatty acid composition of plasma and erythrocyte membranes and was accompanied by a significant decrease of the adhesion molecule α‐4 integrin (CD49d) on lymphocytes but not on monocytes.
Pathological, Clinical and Mycological Findings in Experimental Aspergillosis Infections of Starlings
Authors: Atasever A. , Gümüşsoy K. S.
Published: April 2004

A group of 24 wild starlings (Sturnus vulgaris), in undefined age categories but at least post‐pubertal, constituted the material of this study. Six starlings were kept as a control group and 18 starlings served as the infection group. The starlings in the infection group were infected with inoculums of 1.35 × 106/0.2 ml Aspergillus fumigatus via intratracheal route whereas the control group was administered only placebo in the same way. Six, four, two, four and two birds died on 2, 3, 4, 5 and 6 days post inoculation respectively. At the necropsy of the dead birds, caseous foci were determined in the lungs, on the air sacks, myocardium, thoracic wall and abdominal serosa. In the histopathological examination of the white‐yellowish caseous foci ranging from pinhead to chick pean in size, necrotic granulomatous foci consisting of macrophages, heterophil leukocytes and gigant cells encapsulated with a fibrous tissue were observed. Hyphae and spores of A. fumigatus were determined in these foci using the Gridley staining method.
MMP‐2 activation and stepwise progression of pulmonary adenocarcinoma: Analysis of MMP‐2 and MMP‐9 with gelatin zymography
Authors: Iijima Tatsuo , Minami Yuko , Nakamura Naoko , Onizuka Masataka , Morishita Yukio , Inadome Yukinori , Noguchi Masayuki
Published: April 2004

Small pulmonary adenocarcinomas can be classified on the basis of their histological characteristics and prognosis, and when classified as such, the prognosis of replacing‐type adenocarcinoma with active fibroblast proliferation is significantly worse than adenocarcinoma without fibroblast proliferation. In order to clarify the biological mechanisms of the key to the morphological changes associated with active fibroblast proliferation, we examined the activities of matrix metalloproteinase (MMP)‐2 and MMP‐9, which are important enzymes in the stromal invasion by cancers. The active MMP‐2 and MMP‐9 content of 40 pulmonary adenocarcinomas that were less than 20 mm in diameter was measured by the gelatin zymography method. The quantity of active MMP‐2 in the pulmonary adenocarcinomas with active fibroblast proliferation was higher than in the pulmonary adenocarcinomas without proliferation (P < 0.001), but there were no correlations between the histological features and the activation of MMP‐9. The presence of active fibroblast proliferation in small pulmonary adenocarcinomas suggests that the cancer cells have acquired the ability to invade through the action of active MMP‐2, and this is thought to be one of the reasons for the worse prognosis of pulmonary adenocarcinoma with active fibroblast proliferation.
Distribution of tumor‐infiltrating dendritic cells in human non‐small cell lung carcinoma in relation to apoptosis
Authors: Kurabayashi Atsushi , Furihata Mutsuo , Matsumoto Manabu , Hayashi Hideki , Ohtsuki Yuji
Published: April 2004

Host defense mechanisms play important roles in suppressing the development and growth of tumors. It is known that S‐100 protein‐positive immature dendritic cells (S100DC), as antigen presenting cells (APC), and macrophages have roles in the immune responses to tumor growth. Mediators such as nitric oxide are also important in the surveillance against cancer. We examined the distribution of S100DC and CD68‐positive macrophages (CD68MØ) immunohistochemically to compare the condition of apoptotic tumor cells in 69 patients with human non‐small cell lung carcinoma. The expression of inducible nitric oxide synthases (iNOS) in tumors was also studied. Unlike macrophages, S100DC were distributed predominantly in cancer nests. In the areas with infiltration of ‘many’ S100DC (i.e. more than 10 DC/HPF), we found two distinct patterns of tumor infiltration: scattered and aggregated infiltration of DC in tumor nests. In areas of scattered S100DC distribution, only a few apoptotic tumor cells could be detected. However, in the areas of DC aggregations, apoptotic tumor cells were significantly more abundant (P = 0.0491). In contrast to S100DC, the distribution and density of CD68MØ were associated with iNOS expression of tumor cells (P < 0.0001), but not with distribution of apoptotic tumor cells. These findings reveal differences in the in vivo condition between S100DC and CD68MØ in tumors, and suggest there is a relationship between tumor‐infiltrating S100DC aggregation and apoptosis in in vivo non‐small cell lung cancers.
Adenoid cystic carcinoma with collagenous spherulosis‐like structures in the breast: Report of a case
Authors: Ogata Kentaro , Sakamoto Goi , Sakurai Takashi
Published: April 2004

Adenoid cystic carcinoma (ACC) is rare in breast carcinomas and has a relatively better prognosis than other histological types. Although the tumor shows various growth patterns, such as those of salivary glands or other organs, the papillary growth pattern is uncommon. Collagenous spherulosis (CS), consisting of intraluminal clusters of eosinophilic spherules situated adjacent to, or encompassed by, usually benign proliferative epithelium, is also a rare incidental microscopic finding that ACC must be distinguished from, and it has never been reported in association with ACC. We present a case of ACC arising in mammary ducts with the papillary growth pattern and CS‐like structures that should not be mistaken for benign proliferative lesions, especially in the core needle‐biopsy specimen, because the adenoid cystic pattern might not always be contained in the specimen. Morphologically and histochemically, the results suggest that the CS‐like structures in the present case undergo a similar process to the originally described CS, which the myoepithelium is thought to be associated with. Examination of more cases are necessary to clarify the process of CS and its relationship to mammary proliferative lesions.