bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of differentially expressed genes in a susceptible and moderately resistant potato cultivar upon Phytophthora infestans infection
Authors: Ros Barbara , Thümmler Fritz , Wenzel Gerhard
Published: May 2004

To gain deeper understanding of the host–pathogen interaction in the system potato–Phytophthora infestans, subtractive hybridization in combination with cDNA array hybridization was used. Leaflets of a moderately resistant and a susceptible potato cultivar were inoculated with P. infestans. The infection of the potato leaves was quantified by real‐time quantitative PCR. Using infected and control tissue, two cDNA libraries highly enriched for P. infestans‐induced genes were prepared. Within 531 clones randomly picked and sequenced from the libraries, 285 unigenes were found, from which 182 clones were selected for further analysis by cDNA array hybridization. Sixteen hours post inoculation genes were not induced significantly, whereas 72 h post inoculation induction of gene expression was clearly detectable. In both cultivars, 143 genes were induced moderately (≥ two‐fold), and 35 of the selected genes appeared to be strongly induced (≥ seven‐fold). Among these clones were mainly genes associated with stress and/or defence mechanisms. The strongest gene induction was found in 4‐week‐old susceptible plants. In the moderately resistant cultivar, transcripts of a number of genes accumulate with plant age; as a result, induction of gene expression upon infection was less pronounced. Down‐regulation of three genes was observed in both cultivars, upon infection. Transcript levels of these three genes increased in uninfected plants within 4 weeks of growth. Other differences in defence responses of the two cultivars could be determined and their effects are discussed.
Advances in understanding recessive resistance to plant viruses
Authors: DIAZ‐PENDON JUAN A. , TRUNIGER VERONICA , NIETO CRISTINA , GARCIA‐MAS JORDI , BENDAHMANE ABDELHAFID , ARANDA MIGUEL A.
Published: May 2004

Recent work carried out to characterize recessive mutations which render experimental hosts non‐permissive to viral infection (loss‐of‐susceptibility mutants) seems to be converging with new data on natural recessive resistance in crop species, and also with functional analyses of virus avirulence determinants. Perhaps the most well known examples are the studies that identified the eukaryotic translation initiation factors 4E(iso) (eIF(iso)4E) and 4E(eIF4E) as the host factors required for potyvirus multiplication within experimental and natural hosts, respectively, and the potyviral genome‐linked protein (VPg) as the viral factor that directly interacts with eIF4E to promote potyvirus multiplication. The purpose of this paper is to review the available information on the characterization of loss‐of‐susceptibility mutants in experimental hosts, natural recessive resistances and virus avirulence factors, and also to comment on possible implications for the design of new sources of sustainable virus resistance.
Measurement of cardiac output by transpulmonary arterial thermodilution using a long radial artery catheter. A comparison with intermittent pulmonary artery thermodilution
Authors: L'E. Orme R. M. , Pigott D. W. , Mihm F. G.
Published: May 2004

Cardiac output can be measured accurately by transpulmonary arterial thermodilution using the PiCCO® (Pulsion Medical Systems, Munich, Germany) system with a femoral artery catheter. We have investigated the accuracy of a new 50 cm 4 French gauge radial artery catheter and the ability to use the system with a shorter radial catheter. We studied 18 patients who had undergone coronary artery surgery and made three simultaneous measurements of cardiac output by arterial thermodilution and with a pulmonary artery catheter. The radial catheter was withdrawn in 5 cm increments and the measurements were repeated. We found close agreement between arterial thermodilution and pulmonary artery thermodilution with a mean (SD) bias of 0.38 (0.77) l.min−1. Arterial thermodilution became unreliable once the catheter had been withdrawn by more than 5 cm. We conclude that cardiac output measurement with arterial thermodilution with a radial catheter is interchangeable with that derived from a pulmonary artery catheter, and that a centrally sited arterial catheter is required for accurate determination of cardiac output by transpulmonary arterial thermodilution.
The role of prophylactic fresh frozen plasma in decreasing blood loss and correcting coagulopathy in cardiac surgery. A systematic review
Authors: Casbard A. C. , Williamson L. M. , Murphy M. F. , Rege K. , Johnson T.
Published: May 2004

Fresh frozen plasma is commonly used in cardiac surgery in an attempt to replace clotting factors and to decrease bleeding. Despite this, there has been no previous review of the available literature to support this practice. The aim of this review was to study the effect of prophylactic peri‐operative transfusion of fresh frozen plasma on bleeding and coagulopathy in patients undergoing cardiac surgery. A comprehensive literature search was performed and all randomised controlled trials of the use of fresh frozen plasma in cardiac surgery were included. Six small trials were found that included a total of 363 participants with six different dose regimens of fresh frozen plasma. The overall quality of the studies was poor due to small patient numbers and lack of allocation concealment. There was no evidence that the prophylactic use of fresh frozen plasma affected peri‐operative blood loss in cardiac surgery. There was some evidence that it may improve platelet count and fibrinogen concentration.
Ventilation of a model lung using various cricothyrotomy devices*
Authors: Craven R. M. , Vanner R. G.
Published: May 2004

In this study we developed a model lung to compare the effectiveness of ventilation using four different cricothyrotomy devices. The Ravussin 13G cannula (VBM Medical), the Quicktrach cannula 4 mm ID (VBM Medical), the Melker cannula 6 mm ID (Cook) and a cuffed tracheal tube 6 mm ID were used in turn to ventilate the model lung through a cricothyrotomy over a range of upper airway resistances. The 6 mm cuffed tracheal tube provided consistently good ventilation independent of upper airway resistance. The 6 mm ID Melker device provided at least reasonable and at best very good ventilation, whatever the patency of the upper airway. The Ravussin cannula could ventilate well with the jet ventilator with low upper airway resistance but could not ventilate at all with complete upper airway obstruction. The Quicktrach performed poorly with low upper airway resistance but well with increased upper airway resistance. With its easier insertion, fewer complications compared to a surgical cricothyrotomy, and the ability to use it with a standard anaesthetic circuit, the authors feel that the 6 mm Melker canula is the technique of choice for emergency trans‐tracheal ventilation.
The effects of intrathecal diamorphine on gastric emptying after elective Caesarean section*
Authors: King H. , Barclay P.
Published: May 2004

This study investigated whether intrathecal diamorphine affects gastric emptying following elective Caesarean section. Forty women were randomly allocated to receive either diamorphine 300 μg or 0.9% saline as part of a standard spinal anaesthetic. Gastric emptying was measured in the immediate postoperative period using paracetamol absorption. Paracetamol concentrations were measured by enzymatic assay method using a Beckman CX‐7 automated analyser. The time to maximum concentration (Tmax) was statistically longer in the diamorphine group (control 41.8, SD 20.8 min; diamorphine 72.6 SD 41.9 min; p < 0.01). During the 2‐h study period, mean morphine consumption via a patient controlled analgesia device was significantly higher in the control group (control 9.3, SD 3.6 mg; diamorphine 2.1, SD 2.1 mg; p < 0.01). We conclude that intrathecal diamorphine may contribute to the delay in gastric emptying that occurs immediately following elective spinal Caesarean section.
The impact of regional anaesthesia on outcome: a patient's perspective
Authors: Cornish P. B. , Barton J. , Deacon A.
Published: May 2004

Peripheral nerve catheters are being used increasingly to manage acute pain. Whilst acknowledged as effective, their broader implications for patient outcome are less clear. In this case report, we describe the way in which not only was the pain management of an ischaemic leg successful via a sciatic nerve catheter, but decision‐making around this strategy affected outcome.
Dead space and paediatric anaesthetic equipment: a physical lung model study*
Authors: Miller D. M. , Adams A. P. , Light D.
Published: May 2004

Untested assumptions have been made with regard to functional dead space in facemasks, filters and breathing systems used in children for the administration of inhalation anaesthesia. Total functional dead space was measured in various combinations of this equipment applied to a spontaneous ventilation lung model with parameter settings appropriate for infants of 7–8 kg. We found that functional dead space was too large to allow for spontaneous ventilation of the lungs when a breathing filter was fitted. There was minimal relationship between size of the facemask and functional dead space; however, the provision of 22‐mm female inlets to facemasks achieved proportionately less functional dead space than with 15‐mm male inlets. Regardless of the apparatus used and the magnitude of the dead space, the leak induced when a poorly fitting facemask was used dramatically reduced the dead space of the breathing system – to near optimal conditions – by moving the alveolar gas elimination point to within the facemask itself.
Stability of cultured dental follicle cells
Authors: Yao Shaomian , Norton Jolanna , Wise Gary E.
Published: May 2004

Abstract.  Because the dental follicle is required for tooth eruption, establishment of dental follicle cell (DFC) lines is needed for experimentation to determine how the cells regulate eruption. Thus, it is critical that the follicle cells in culture remain stable and neither become transformed nor differentiate. To determine the stability of rat DFC cultures in terms of exhibiting contact inhibition of growth when confluent (no transformation), DFC at different passages were analysed using flow cytometry. Gene expression of cyclin E was determined by reverse transcription polymerase chain reaction as a further method to determine if growth was occurring when the cells were confluent. Alkaline phosphatase and von Kossa staining were also performed as a means of determining stability in terms of differentiation; that is, are the DFC maintaining their phenotype or are they differentiating into osteoblasts and osteocytes? After plating cells of a given passage, they initially underwent a rapid phase of growth with 30–40% of the cells in S, G2 and M (dividing track) as determined by flow cytometry. The number of such cells declined to only 7–15% at preconfluency. At late confluency, only 2 and 5% of the cells were in the dividing track in passages 6 and 9, respectively, but in passage 12 this had risen to 15%. For a given passage of cells, cyclin E gene expression significantly declined in late confluency as compared to the early growth phase. However, in passage 12, the gene expression of cyclin E at late confluency was higher than the expression at late confluency in passage 6. Thus, the DFC were remarkably stable through passage 9, but by passage 12 it appeared that a small percentage of the cells had become transformed and had lost their contact inhibition growth properties. Alkaline phosphatase and von Kossa staining were negative for all passages, suggesting that the cells remained stable in terms of differentiation and did not differentiate into either osteoblasts or osteocytes.
Human airway submucosal glands augment eosinophil chemotaxis during rhinovirus infection
Authors: Furukawa E. , Ohrui T. , Yamaya M. , Suzuki T. , Nakasato H. , Sasaki T. , Kanda A. , Yasuda H. , Nishimura H. , Sasaki H.
Published: May 2004

Background
Asthma exacerbations are frequently associated with rhinovirus (RV) infections. However, the contribution of airway submucosal gland (SMG) to exacerbations of asthma in RV respiratory infection has not been studied.

Objective
This study was undertaken to examine whether RV‐infected human respiratory SMG cells produce pro‐inflammatory cytokines and chemokines for eosinophils, and augment eosinophil transmigration across human airway epithelium.

Methods
We infected cultured human tracheal SMG cells with RV14, collected culture media at 1, 3, and 5 days after infection, and measured the chemotactic activity for eosinophils in the culture supernatant using a 48‐well microchemotaxis chamber and a 51Cr‐labelled eosinophil transmigration assay.

Results
Exposing a confluent human tracheal SMG cell monolayer to RV14 consistently led to infection. Human SMG cells with RV infection secreted soluble factors activating human eosinophil chemotaxis into the culture supernatant in a time‐dependent manner, and the culture supernatant significantly augmented the transmigration of 51Cr‐labelled eosinophils through human airway epithelial cell layers from the basal to mucosal side. These effects were completely abolished by a mixture of a monoclonal antibody regulated on activation, normal T cells expressed and secreted (RANTES) and an antibody to granulocyte macrophage‐colony stimulating factor (GM‐CSF).

Conclusion
These results suggest that human respiratory SMG cells may augment eosinophil transmigration across the airway epithelium through the secretion of RANTES and GM‐CSF after RV infection, and may contribute to exacerbations of asthma.
Does previous infection protect against atopic eczema and recurrent wheeze ininfancy?
Authors: Kramer M. S. , Guo T. , Platt R. W. , Sevkovskaya Z. , Dzikovich I. , Collet J.‐P. , Shapiro S. , Chalmers B. , Hodnett E. , Vanilovich I. , Mezen I. , Ducruet T. , Shishko G. , Bogdanovich N.
Published: May 2004

Background
Frequent infection in infancy and early childhood has been hypothesized to explain the low prevalence of asthma and other atopic disease among children in developing countries (the so‐called ‘hygiene hypothesis’), but the low prevalence in Eastern Europe remains unexplained.

Objective
To test the hygiene hypothesis in the Republic of Belarus by examining the relationship between gastrointestinal (GI) and respiratory infection and two potentially atopic outcomes in the first 12 months of life: atopic eczema and recurrent wheeze.

Methods
We carried out two case–control studies nested within a large (n=17 046) randomized trial in Belarus, with cases defined as (1) first occurrence of atopic eczema (n=819) and (2) second episode of wheezing (n=112). Incidence density sampling was used to select four matched controls born within 1 month at the same hospital as the case. Exposure was defined as one or more episodes of GI or respiratory infection (examined separately) with onset >7 days before onset of the case's atopic outcome. Analyses controlled for family atopic history, duration of exclusive breastfeeding, sex, birth weight, maternal education, and (for recurrent wheeze) maternal smoking.

Results
For atopic eczema, prior GI infection occurred in 7.4% of cases vs. 6.0% of controls [adjusted OR=1.27 (0.94–1.72)] and prior respiratory infection in 35.2% vs. 32.6% [adjusted OR=1.14 (95% CI=0.94–1.37)]. For recurrent wheeze, prior GI infection occurred in 9.8% of cases vs. 7.4% of controls [adjusted OR=1.30 (0.60–2.82)].

Conclusion
Our results do not support the hypothesis that infection protects against atopic eczema or recurrent wheezing in the first 12 months of life.
A novel promoter polymorphism in the gene encoding complement component 5 receptor 1 on chromosome 19q13.3 is not associated with asthma and atopy in three independent populations
Authors: Barnes K. C. , Caraballo L. , Muñoz M. , Zambelli‐Weiner A. , Ehrlich E. , Burki M. , Jimenez S. , Mathias R. A. , Stockton M. L. , Deindl P. , Mendoza L. , Hershey G. K. , Nickel R. , Wills‐Karp M.
Published: May 2004

Background
The inflammatory functions of complement component 5 (C5) are mediated by its receptor, C5R1, which is expressed on bronchial, epithelial, vascular endothelial and smooth muscle cells. A susceptibility locus for murine allergen‐induced airway hyper‐responsiveness was identified in a region syntenic to human chromosome 19q13, where linkage to asthma has been demonstrated and where the gene encoding C5R1 is localized.

Objective
The aim of this study was to screen for novel polymorphisms in the C5R1 gene and to determine whether any identified polymorphisms are associated with asthma and/or atopy and whether they are functional.

Methods
Single‐nucleotide polymorphism (SNP) detection in the gene encoding C5R1 was performed by direct sequencing. Genotyping was performed in three populations characterized for asthma and/or atopy: (1) 823 German children from The Multicenter Allergy Study; (2) 146 individuals from Tangier Island, Virginia, a Caucasian isolate; and (3) asthma case–parent trios selected from 134 families (N=783) in Barbados. Functional studies were performed to evaluate differences between the wild‐type and the variant alleles.

Results
We identified a novel SNP in the promoter region of C5R1 at position −245 (T/C). Frequency of the −245C allele was similar in the German (31.5%) and Tangier Island (36.3%) populations, but higher in the Afro‐Caribbean population (53.0%; P=0.0039 to <0.0001). We observed no significant associations between the −245 polymorphism and asthma or atopy phenotypes. Upon examination of the functional consequences of the −245T/C polymorphism, we did not observe any change in promoter activity.

Conclusion
This new marker may provide a valuable tool to assess the risk for C5a‐associated disorders, but it does not appear to be associated with asthma and/or atopy.
Roles of major oligosaccharides on Cry j 1 in human immunoglobulin E and T cell responses
Authors: Okano M. , Kimura Y. , Kino K. , Michigami Y. , Sakamoto S. , Sugata Y. , Maeda M. , Matsuda F. , Kimura M. , Ogawa T. , Nishizaki K.
Published: May 2004

Background
We have demonstrated that carbohydrates in Cry j 1, the major allergen of Cryptomeria japonica pollen, play a major role in promoting Cry j 1‐specific Th2 response. However, little is known as to whether the carbohydrates directly participate in allergic responses.

Objective
We sought to determine whether Cry j 1‐related oligosaccharides function as IgE and/or T cell epitopes. In addition, the regulatory effect of Cry j 1‐related oligosaccharide on Cry j 1‐specific T cell responses was investigated.

Methods
Two monovalent oligosaccharides largely found on Cry j 1, Manα1‐6(Manα1‐3)(Xylβ1‐2)Manβ1‐4GlcNAcβ1‐4(Fucα1‐3)GlcNAc (M3FX), and GlcNAcβ1‐2Manα1‐6(GlcNAcβ1‐2Manα1‐3)(Xylβ1‐2)Manβ1‐4GlcNAcβ1‐4(Fucα1‐3)GlcNAc (GN2M3FX) were prepared. Manα1‐2Manα1‐6(Manα1‐2Manα1‐3)Manα1‐6(Manα1‐2Manα1‐2Manα1‐3)Manβ1‐4GlcNAcβ1‐4GlcNAc (M9A) was used as control. Competitive inhibition ELISA for Cry j 1‐specific IgE was performed using these oligosaccharides as inhibitors. In addition, T cell lines specific for Cry j 1 or purified protein derivative of Mycobacterium tubecurosis (PPD) were established, and cellular responses against these oligosaccharides were investigated in the presence or absence of the respective antigens.

Results
Overall, neither M3FX nor GN2M3FX displayed inhibitory effect on the binding between IgE and Cry j 1. In addition, M3FX did not by itself stimulate Cry j 1 or PPD‐specific T cells. However, M3FX significantly inhibited Cry j 1‐induced proliferation and IL‐4 production in Cry j 1‐specific T cells. Such an inhibitory effect was not seen in PPD‐specific T cell responses.

Conclusion
These results suggest that Cry j 1‐related oligosaccharides are not major epitopes for IgE or T cells. However, these oligosaccharides have a novel potential to inhibit Cry j 1‐specific T cell responses selectively.
No apparent association between periocular and ocular microcolonization and the degree of inflammation in patients with atopic keratoconjunctivitis
Authors: Nivenius E. , Montan P. G. , Chryssanthou E. , Jung K. , Van Hage‐Hamsten M. , Van Der Ploeg I.
Published: May 2004

Background
The cause of the chronic inflammation in atopic keratoconjunctivitis (AKC), the ocular manifestation of atopic eczema/dermatitis syndrome, is largely unknown.

Objective
To investigate the possibility that microorganisms may be important in the inflammatory activity in AKC.

Methods
Fifteen patients with AKC participated in the study. The presence of aerobic bacteria and fungi was related to the severity of clinical signs, the numbers of inflammatory cells in tears and conjunctival biopsies, and the concentration of various cytokines in tears. In addition, serological evidence for IgE sensitization to Staphylococcus aureus B antigen and Malassezia sympodialis antigen was investigated. Twelve healthy subjects were included for control purposes.

Results
The patients exhibited moderate clinical signs of AKC. No relation was found between the severity of AKC and the presence of microorganisms, despite the fact that S. aureus was frequently isolated. AKC patients showed significantly higher levels of IFN‐γ, TNF‐α (tumour necrosis factor‐α), IL‐2, IL‐4, IL‐5 and IL‐10 than controls. An association was found between conjunctival signs and the levels of all cytokines except IL‐5.

Conclusion
We found no evidence to suggest that periocular and ocular microcolonization are related to inflammatory parameters in AKC. However, confirmation of the present results in a longitudinal study with repeated clinical examinations and samplings in the same individual is required before the contribution of S. aureus to on‐going inflammation in AKC can be dismissed.
Late eczematous reactions to food in children with atopic dermatitis
Authors: Breuer K. , Heratizadeh A. , Wulf A. , Baumann U. , Constien A. , Tetau D. , Kapp A. , Werfel T.
Published: May 2004

Background
Food allergy is a common problem in patients with atopic dermatitis (AD), particularly in children. While immediate reactions to food are well characterized, the importance of food as a provocation factor for late eczematous reactions has been a subject of debate for several decades.

Objective
To investigate the importance of food for the induction of late eczematous reactions in children with AD and to correlate the clinical outcome to the results of specific IgE determinations and atopy patch tests (APTs).

Methods
One hundred and six double‐blind placebo‐controlled food challenges (DBPCFCs) to cow's milk, hen's egg, wheat and soy in 64 children with AD (median age 2 years) were analysed retrospectively. Total and food‐specific IgE were determined by CAP RAST FEIA and APTs with native foodstuff were performed. The diagnostic values of specific IgE and APT results were calculated.

Results
Forty‐nine (46%) of the challenges were related to a clinical reaction. An exacerbation of AD (late eczematous reaction) commonly occurred 24 h after the ingestion of food. Isolated late eczematous reactions were seen in 12% of all positive challenges. Forty‐five percent of the positive challenges were associated with late eczematous responses, which followed immediate‐type reactions. The sensitivity of food‐specific IgE and the APT was 76% and 70%, respectively. Specific IgE and APT were often false positive, which resulted in low positive predictive values (64% and 45%, respectively).

Conclusions
Late eczematous reactions may often be observed upon food challenge in children with AD. Due to the poor reliability of food‐specific IgE and APT results DBPCFCs have still to be regarded as the gold standard for the appropriate diagnosis of food responsive eczema in children with AD.
Induction of eotaxin production by interleukin‐4, interleukin‐13 and lipopolysaccharide by nasal fibroblasts
Authors: Nonaka M. , Pawankar R. , Fukumoto A. , Ogihara N. , Sakanushi A. , Yagi T.
Published: May 2004

Background
There is growing evidence that eotaxin is a key mediator in the development of tissue eosinophilia. Fibroblasts are a major source of eotaxin. The severity of diseases with eosinophilic inflammation like nasal polyposis, atopic dermatitis and asthma, where Th2‐type cytokines (IL‐4 and IL‐13) and TGF‐β are expressed locally, was shown to correlate with bacterial factors such as lipopolysaccharide (LPS) rather than allergen.

Objective
We examined eotaxin production by nasal fibroblasts stimulated with IL‐4 or IL‐13 alone or in combination with LPS, and the effect of TGF‐β1 on it. Moreover, we compared the magnitude of eotaxin produced by nasal fibroblasts with that produced by lung or skin fibroblasts.

Methods
Fibroblast lines were established from human biopsy tissue. The expression of eotaxin mRNA was evaluated by RT‐PCR. The amount of eotaxin in the supernatants was measured by ELISA.

Results
IL‐4, but not IL‐13, synergized with LPS to produce eotaxin in a dose‐ and time‐dependent manner. Sequential treatment of nasal fibroblasts with IL‐4 and LPS did not have any effect. But when IL‐4 and LPS were added together, synergy for eotaxin production was observed. Moreover, this synergy was observed in nasal and skin fibroblasts, but not in lung fibroblasts. The production of eotaxin by IL‐4 and LPS was modulated by TGF‐β1.

Conclusion
These results suggest that a co‐stimulus like LPS is necessary for IL‐4 to make a strong induction of eotaxin in eosinophilic inflammations such as nasal polyposis. Modulation by TGF‐β1 may have important implications for the development of eosinophilic inflammation.
Association of HLA‐DQ5 and HLA‐DR1 with sensitization to organic acidanhydrides
Authors: Jones M. G. , Nielsen J. , Welch J. , Harris J. , Welinder H. , Bensryd I. , Skerfving S. , Welsh K. , Venables K. M. , Taylor A. N.
Published: May 2004

Background
Organic acid anhydrides are low molecular weight industrial chemicals, able to cause rhinoconjunctivitis and asthma associated with specific IgE against hapten–carrier protein conjugate. Only a proportion of exposed workers develop IgE‐associated allergy to acid anhydrides.

Objective
We determined whether genetic susceptibility, in particular, HLA Class II alleles may be a risk factor.

Methods
We undertook HLA typing in 52 cases who had confirmed specific IgE and in 73 referents matched on site, age and duration of acid anhydride exposure identified in cross‐sectional studies of workers exposed to hexahydrophthalic (HHPA), methylhexahydrophthalic (MHHPA) and methyltetrahydrophthalic (MTHPA) anhydrides.

Results
The linked alleles DQ5 (odds ratio [OR]=4.3; 95% confidence interval [95% CI]=1.7, 11) and DR1 (OR 3.0; 95% CI 1.2, 11) were more prevalent in cases than in referents. Within DQ5, DQB1*0501 was particularly frequent (OR 3.0; 95% CI 1.2, 7.4).

Conclusion
DQB1*05 gene confers susceptibility to develop specific IgE antibodies against HHPA, MHHPA and a non‐significant trend with MTHPA. DQB1*0501 is protective for other low molecular chemical sensitizers (isocyanates and plicatic acid) which may indicate varying affinities for the corresponding specific class II molecules.
Identification of systemically expanded activated T cell clones in MRL/lpr and NZB/W F1 lupus model mice
Authors: ZHOU G , FUJIO K , SADAKATA A , OKAMOTO A , YU R , YAMAMOTO K
Published: May 2004

CD4+ T lymphocytes play an important role in the pathogenesis of systemic lupus erythematosus (SLE). To characterize the clonal expansion of CD4+ T cells in murine lupus models, we analysed the T cell clonality in various organs of young and nephritic MRL/lpr and NZB/W F1 mice using reverse transcription–polymerase chain reaction (RT-PCR) and subsequent single-strand conformation polymorphism (SSCP) analysis. We demonstrated that some identical T cell clonotypes expanded and accumulated in different organs (the bilateral kidneys, brain, lung and intestine) in nephritic diseased mice, and that a number of these identical clonotypes were CD4+ T cells. In contrast, young mice exhibited little accumulation of common clones in different organs. The T cell receptor (TCR) Vβ usage of these identical clonotypes was limited to Vβ2, 6, 8·1, 10, 16 and 18 in MRL/lpr mice and to Vβ6 and 7 in NZB/W F1 mice. Furthermore, some conserved amino acid motifs such as I, D or E and G were observed in CDR3 loops of TCRβ chains from these identical CD4+ clonotypes. The existence of systemically expanding CD4+ T cell clones in the central nervous system (CNS) suggests the involvement of the systemic autoimmunity in CNS lesions of lupus. FACS-sorted CD4+CD69+ cells from the kidney displayed expanded clonotypes identical to those obtained from the whole kidney and other organs from the same individual. These findings suggest that activated and clonally expanded CD4+ T cells accumulate in different tissues of nephritic lupus mice, and these clonotypes might recognize restricted T cell epitopes on autoantigens involved in specific immune responses of SLE, thus playing a pathogenic role in these lupus mice.
Decrease of CD154 intensity on peripheral CD4+ T cells in autoimmune thyroid disease
Authors: WATANABE M , YAMAMOTO N , MATSUZUKA F , MIYAUCHI A , IWATANI Y
Published: May 2004

To clarify immunological differences among patients with Graves’ disease (GD) and Hashimoto's disease (HD) at various levels of severity, we examined the expression of the CD154 molecules on peripheral T cells, which regulate B cell activation, B cell differentiation, and T-cell survival. We found decreases in the intensities of CD154 on peripheral CD4+ cells from euthyroid patients with GD and HD, but we did not find any differences between patients with different disease severities. CD8+ cells did not express CD154 molecules. Thus, CD154 expression on CD4+ cells may be related to the pathogenesis of the autoimmune thyroid diseases, not to the disease severity.
Impaired chemokine‐induced migration during T‐cell development in the absence of Jak 3
Authors: Soldevila Gloria , Licona Ileana , Salgado Alfonso , Ramírez Marcela , Chávez Ramsés , García‐Zepeda Eduardo
Published: May 2004

The arrival of bone marrow T‐cell progenitors to the thymus, and the directed migration of thymocytes, are thought to be regulated by the expression of chemokines and their receptors. Recent data has shown that the Jak\Stat signalling pathway is involved in chemokine receptor signalling. We have investigated the role of Jak 3 in chemokine‐mediated signalling in the thymus using Jak 3–\– mice. These mice show defects in T‐cell development, as well as in peripheral T‐cell function, resulting in a hypoplastic thymus and an altered T‐cell homeostasis. Here we demonstrate, for the first time, that bone marrow progenitors and thymocytes from Jak 3–\– mice have decreased chemotactic responses to CXCL12 and CCL25. We also show that Jak 3 is involved in signalling through CCR9 and CXCR4, and that specific inhibition of Jak 3 in wild‐type progenitors and thymocytes decreases their chemotactic responses towards CCL25 and CXCL12. Finally, quantitative reverse transcription–polymerase chain reaction analysis showed that thymocytes from Jak 3–\– mice express similar levels of CXCR4 and CCR9 compared to wild‐type mice. Altogether, deficient CCL25‐ and CXCL12‐induced migration could result in a homing defect of T‐cell progenitors to the thymus, as well as in a deficient thymocyte migration through the thymic stroma. Our results strongly suggest that the absence of Jak 3 affects T‐cell development, not only through an impaired interleukin‐7 receptor (IL‐7R)‐mediated signalling, but also through impaired chemokine‐mediated responses, which are crucial for thymocyte migration and differentiation.
Classical and alternative pathway complement activation are not required for reactive systemic AA amyloid deposition in mice
Authors: Hutchinson Winston L. , Herbert Jeff , Botto Marina , Pepys Mark B.
Published: May 2004

During induction of reactive systemic amyloid A protein (AA) amyloidosis in mice, either by chronic inflammation or by severe acute inflammation following injection of amyloid enhancing factor, the earliest deposits form in a perifollicular distribution in the spleen. Because the splenic follicular localization of immune complexes and of the scrapie agent are both complement dependent in mice, we investigated the possible complement dependence of AA amyloid deposition. In preliminary experiments, substantial depletion of circulating C3 by cobra venom factor had little effect on experimental amyloid deposition. More importantly, mice with targeted deletion of the genes for C1q or for both factor B and C2, and therefore unable to sustain activation, respectively, of either the classical complement pathway or both the classical and alternative pathways, showed amyloid deposition similar to wild type controls. Complement activation by either the classical or alternative pathways is thus not apparently necessary for the experimental induction of systemic AA amyloid in mice.
The mechanism for activation of the neutrophil NADPH‐oxidase by the peptides formyl‐Met‐Leu‐Phe and Trp‐Lys‐Tyr‐Met‐Val‐Met differs from that for interleukin‐8
Authors: Fu Huamei , Bylund Johan , Karlsson Anna , Pellmé Sara , Dahlgren Claes
Published: May 2004

Neutrophil chemotaxis has been shown to be regulated by two different signalling pathways that allow strong chemoattractants, such as bacterial‐derived formylated peptides, to dominate over endogenous attractants, such as interleukin‐8 (IL‐8). Here we show that triggering of the formyl peptide receptor (FPR) with f‐Met‐Leu‐Phe (fMLF) substantially reduced the neutrophil superoxide production induced by activation of the CXC receptors with IL‐8. When the order of agonists was reversed, the cells were primed in their response to fMLF, suggesting that the signalling hierarchy between strong, so‐called end‐type (i.e. fMLF) and weak or intermediate‐type (i.e. IL‐8) chemoattractants, is also operating during activation of the NADPH‐oxidase. The same result was obtained when fMLF was replaced with the hexapeptide, WKYMVM, specific for the formyl peptide‐like receptor 1 (FPRL1). There were additional differences between the agonist receptor pairs fMLF/FPR, WKYMVM/FPRL1 and IL‐8/CXCR. In contrast to FPR and FPRL1, no reserve pool of CXCR was present in subcellular granules and it was impossible to prime the oxidative response transduced through CXCR by the addition of priming agents such as tumour necrosis factor‐α and platelet‐activating factor. Moreover, the cytoskeleton‐disrupting substance, cytochalasin B, had no effect either on IL‐8‐triggered oxidase activation or on CXCR reactivation. A pertussis toxin‐sensitive G‐protein is involved in signalling mediated through both FPR and CXCR, and the signalling cascades include a transient intracellular calcium increase, as well as downstream p38 MAPK and phosphoinositide 3‐kinase activation. The data presented in this study provide support for two different signalling pathways to the neutrophil NADPH‐oxidase, used by ligand binding to FPR/FPRL1 or CXCR, respectively.
A Histological Study of the hph‐1 Mouse Mutant: an Animal Model of Phenylketonuria and Infantile Hypertrophic Pyloric Stenosis
Authors: Abel R. M. , Dorè C. J. , Bishop A. E. , Facer P. , Polak J. M. , Spitz L.
Published: May 2004

Aim: To quantify the chronological sequence of changes in the morphology and immunoreactivity for neurotransmitters in the pylorus of an animal model of infantile hypertrophic pyloric stenosis and phenylketonuria. Method: Thirty specimens of pylorus from hph‐1 mice and age/sex matched controls (age range: 10–180 days) were examined using conventional histology and immunohistochemistry for a variety of antigens: protein gene product 9.5, a pan neuronal marker; vasoactive intestinal polypeptide; nitric oxide synthase two antigens coalesced to the same inhibitory neurons in humans; substance P, a potent excitatory neurotransmitter; and calcitonin gene related peptide, a neurotransmitter implicated in the somatic afferent innervation of the stomach. The changes in the morphology of the muscle layers were quantified and statistically analysed for each age group (10, 20, 40, 90 and 180 days). Results: Between 10 and 90 days of age, all muscle layers of the hph‐1 mice were hypertrophied, for example, 10 days, hph‐1 longitudinal muscle mean diameter = 3.4, control = 1.8; hph‐1 circular muscle width = 11.5, control = 4.7. The hph‐1 mice were significantly smaller during this period (40 days, hph‐1 weight = 10 g, control = 25 g). There was no change in the pattern of expression of the antigens examined within the hph‐1 mice compared with the controls. Conclusion: Hph‐1 mice develop a transient smooth muscle hypertrophy of the pylorus attended by gastric distension and failure to gain weight. These changes resolve as the pyloric muscle hypertrophy resolves.
Some Segmental Features on the Structure of the Aortic Wall of the Dog
Authors: Orsi A. M. , Stefanini M. A. , Crocci A. J. , Simões K. , Ribeiro A. A. C. M.
Published: May 2004

Structural features of segmental parts of the aorta of the dog were studied by light microscopy (LM) and scanning electron microscopy (SEM). The variability in the wall architecture composition and vascular thickness of the ascending (T2‐3 level), thoracic (T7‐8 level) and abdominal (L6‐7 infrarenal level) segments of the aorta was analysed. Morphological features such as presence of intimal folds, pattern of the medial myoconnective components with segmental variations in the number of elastic lamellae, whose relative number was higher in the thoracic aorta (ascending and descending parts), compared with the abdominal aorta, and a network of connective (stromal) elements formed by elastic and collagen lamellae and fibres in the adventitia were observed. The results were discussed on a histophysiological basis, because small but significant segmental differences had been characterized in the aortic wall structure of the dog.
Immunocytochemical Detection of Synaptophysin in Enteric Neurones during Prenatal Development in the Rat Stomach
Authors: Asar M. , Bayram Z. , Korgun E. T. , Tertemiz F. , Akkoyunlu G. , Demir R.
Published: May 2004

In this study, the localization and appearance of synaptophysin‐immunoreactive (IR) nerve cells and their relationships with the developing gastric layers were studied by immunocytochemistry and light microscopy in the embryonic rat stomach. The stomachs of Wistar rat embryos aged 13–21 days were used. The first neuronal bodies and their processes containing synaptophysin‐immunoreactivity were observed on embryonic day 13. In contrast, synaptophysin‐IR nerve terminals were first observed between mesenchymal cells on embryonic day 14. These results indicate that synaptophysin is expressed in growing neurits and neuronal cell bodies before these neurones have established synaptic connections. The occurrences of mesenchymal cell condensation near synaptophysin‐IR neuroblasts on embryonic day 15 reflect an active nerve element‐specific mesenchymal cell induction resulting in the morphogenesis of muscle cells. Similarly, the appearance of glandular structures after synaptophysin‐IR neuroblasts, on embryonic day 18, suggests that the epithelial differentiation may be closely related to the neuronal maturation as well as other factors. Finally, synaptophysin is functionally important in neuronal development and maturation, together with the establishment of neuroneuronal and neuromuscular contacts and in epithelial differentiation.
Histo‐anatomical Studies on the Accessory Reproductive Glands of the Anatolian Souslik (Spermophilus xanthoprymnus) (Mammalia: Sciuridae)
Authors: Çakir M. , Karataş A.
Published: May 2004

In this study, the histo‐anatomy of the accessory reproductive glands of the Anatolian souslik (Spermophilus xanthoprymnus) is investigated, using 102 males of the vouchers which were captured from central Anatolia and eastern Mediterranean regions. Averages of measurements taken from each part of the accessory reproductive glands were as follows: length of vesicular gland [right: 3.96 (±0.48) mm; left: 3.70 (±0.55) mm]; prostate gland [length: 2.55 (±0.38) mm; width: 2.63 (±0.45) mm]; length of bulbourethral gland [right: 2.32 (±0.33) mm; left: 2.37 (±0.39) mm], and the length of the pars pelvina of the urethra was 12.31 (±0.76) mm. Histological sections showed that vesicular gland was composed of typically branched tubulo‐alveolar glands. As the fibrous connective tissue that was found among alveoles in the prostate was dense and relatively common, only few tubulo‐alveolar glands occurred. The bulbourethral gland (Cowper's gland) was a tubulo‐alveolar gland with a large and serrated lumen and septa between the alveoles consisted of dense fibro‐muscular fibres.
Neurosteroidogenesis in Oligodendrocytes and Purkinje Neurones of Cerebellar Cortex of Dogs
Authors: Yarim M. , Kabakci N.
Published: May 2004

The cerebellum is a steroidogenic organ that expresses steroidogenic enzymes and produces neurosteroids. Purkinje neurones appear to be the most active steroidogenic cells in the cerebellar cortex. These neurones express 3β‐hydroxysteroid dehydrogenase (3β‐HSD), P450 side‐chain cleavage (P450scc), 17α‐hydroxylase/c17, 20lyase (P450c17), P450 aromatase (P450arom) and produce pregnenolone, progesterone, dehydroepiandrosterone, androstenedion, oestradion and oestrone. Oligodendrocytes are predominantly the producer of myeline protein. The oligodendrocytes were identified by immunohistochemistry using a monoclonal antibody against myeline 2′, 3′‐cyclic nucleotide 3′‐phosphodiesterase (CNPase), a myeline specific enzyme. In this study we have examined the distribution of 3β‐HSD and CNPase by immunohistochemistry using monoclonal antibody in canine cerebellar cortex. The localization of oligodendrocytes within the cerebellar cortex was determined to be close to Purkinje neurones. This result suggests that endogenous progesterone synthesized de novo in the Purkinje neurone can promote myeline protein synthesis in oligodentrocytes.
Quantitative Analysis of Cells in the Ganglion Cell Layer of the Chick Retina: Developmental Changes in Cell Density and Cell Size
Authors: Chen Y. , Wang Z. , Shibata H. , Naito J.
Published: May 2004

Changes in cell density and size in the ganglion cell layer (GCL) of the retina were studied in chick embryos and post‐hatching chicks. The total number of cells in the GCL increased from 3.64 million at embryonic day 8 (E8) to the maximal 7.85 million at E14. After E14, the number of cells decreased to 6.08 million at post‐hatching day 1 (P1) and 4.87 million at P8. Cell density in the GCL decreased unevenly according to retinal regions; cell density in the presumptive central area (pCA) of P8‐chicks decreased to approximately 45% of that in E8‐embryos. Densities of the nasal peripheral retina (NP) and temporal peripheral retina (TP) of P8‐chicks decreased to 23 and 18% of E8‐embryos, respectively. Differentiation of the central (44 000 cells/mm2 in pCA) – peripheral (28 000 cells/mm2 in TP) gradient in cell density was formed by E8. The presumptive dorsal area (pDA) was shaped by E11, but became obscure with age. Although ganglion cell sizes were basically uniform at E8, differentiation occurred with the appearance of larger ganglion cells after E14. Mean size of retinal ganglion cells increased 2.8‐fold in the pCA and 3.8‐fold in the TP between E8 and P8, accompanying a similar scale of decreases in cell densities.
Stereology of the Liver in Three Species ofLeontopithecus(Lesson, 1840) Callitrichidae – Primates
Authors: Burity C. H. F. , Pissinatti A. , Mandarim‐de‐Lacerda C. A.
Published: May 2004

Studies on liver morphology and stereology are relevant to the comparative anatomical and pathological research. They also facilitate the use of non‐human primates in basic research, which has substantially supported studies in human medicine. Quantitative studies of liver structures have also been more extensive in Old World primates and other vertebrates. Twenty‐three livers of adult lion tamarins were studied (sixLeontopithecus rosalia, sevenLeontopithecus chrysomelas, and 10Leontopithecus chrysopygus), dissected, and fixed in 10% neutral buffered formalin solution. For stereological quantification, the liver was regarded as consisting of parenchyma (hepatocytes) and stroma (non‐hepatocytes). The volume density (Vv) was determined by point counting, and the disector method was used to obtain the numerical density of hepatocytes (Nv). Hepatic stereological differences among the three species of lion tamarins were not statistically significant. Therefore, the pooledVv[hepatocyte]andVv[stroma]could be determined as 96.2 and 7.4%, respectively, andNv[hepatocyte]as 500.33 × 106 cm−3. Significantly different, the values found forVv[hepatocyte]andNv[hepatocyte]in lion tamarins were, respectively, 0.09 and 2.8 times greater than those in baboons, and 0.17 and 3.8 times greater than those in man. However, theVv[stroma]was 1.04 times smaller than that in baboons and 1.79 times smaller than that in man.
Influence of Water Deficit on Uptake and Distribution of Nitrogen in Soybeans Monitored by Soil Injected 15N
Authors: Herzog H. , Götz K.‐P.
Published: May 2004

A pot experiment was carried out in a climatic chamber to study 15N‐uptake, translocation and recovery in soybeans [Glycine max (L.) Merr.] as affected by a 13 day interval of water deficits (wd) starting at flowering. Ten or 40 mg 15N (500 or 2000 p.p.m. 15N) was injected at 0.4 m depth into the soil 3 days after flowering (3 DAF). A dose of 10 mg 15N, resulted in a translocation of 50 and 80 % to the shoot at 7 and 13 DAF, respectively; thus the uptake of 15N into the plant organs was limited by the low remaining 15N in the soil after 7 DAF. Therefore, only results referring to the 40 mg 15N application are presented here. The daily 15N‐uptake of well‐water (ww) plants during the 10 days of investigations was about c. 2.5 mg 15N, it was significantly higher than under wd at 7 DAF and tended to be higher at 13 DAF. The reduction by wd was also evident by a lower recovery of 15N (55 % instead of 75 %) at 13 DAF. In almost all leaves and pods there was a trend to a lowered translocation (mg 15N per organ) under wd when compared with the ww plants at 7 and 13 DAF; lowering due to wd was strongest and significant in upper leaves and pods at 13 DAF, whereas translocation to stems was even slightly increased. Relative accumulation of 15N (mg g−1 dry matter) in leaves, stems and pods increased from the bottom to the top indicating an higher sink activity of younger organs. Water deficits tended to lower this relative 15N‐accumulation in all leaves and pods at 7 and 13 DAF, and caused a significant reduction in upper leaves and pods at 7 DAF. Absolute and relative 15N‐accumulation in stems tended to be affected differently by wd, being unaffected or slightly decreased at 7 DAF, but increased at 13 DAF. These differential effects of wd are discussed.
Influence of Seeding Depth and Seedbed Preparation on Establishment, Growth and Yield of Fibre Flax (Linum usitatissimum L.) in Eastern Canada
Authors: Couture S. J. , DiTommaso A. , Asbil W. L. , Watson A. K.
Published: May 2004

Research was conducted at the Macdonald Campus of McGill University (Québec, Canada) at three sites in 1997 and one site in 1998 to determine the effects and interactions of seeding depth (0, 1, 2, 4 or 6 cm) and seedbed preparation (i.e. soil rolling): none, rolling before or rolling after seeding on fibre flax (cv. Ariane) establishment, growth and yield. Seedbed preparation had little impact on the parameters measured while seeding depth had a variable effect on plant density, plant height, stem diameter and retted straw yield. Seeding depths of 1–4 cm provided consistently good establishment, growth and yield results. In 1997, there was an interaction between seeding depth and seedbed preparation on plant height, branching ratio and retted straw yield, although results were generally variable and tended to be site‐specific. In 1998, there was an interaction between seeding depth and seedbed preparation on plant height and stem diameter prior to harvest, with the results varying for all seeding depth‐seedbed preparation treatment combinations except for the 2‐cm depth treatment. Rolling of the seedbed before seeding on lighter soils and at a depth of 2 cm on most soils can improve establishment, growth and yields of fibre flax under eastern Canadian growing conditions.