bluuebunny/crossref_metadata_2025 huggingface dataset:
Improvement of semen quality by nocturnal scrotal cooling in oligozoospermic men with a history of testicular maldescent
Authors: JUNG A. , SCHILL W. ‐B. , SCHUPPE H. ‐C.
Published: March 2005

For infertile men with a history of testicular maldescent only few therapeutic options exist beside assisted reproduction. The aim of our study was to evaluate the influence of nocturnal scrotal cooling on semen quality in such patients presenting with oligozoospermia. Twenty infertile men with a history of testicular maldescent and oligozoospermia were included for nocturnal scrotal cooling over 12 weeks for every night. To increase nocturnal periscrotal air circulation we used a membrane pump connected via plastic tubes to receptacles placed in both groins. Semen analysis was performed at the beginning of the cooling period and at weeks 4, 8 and 12. Another 20 infertile patients with a history of testicular maldescent and oligozoospermia were followed without specific treatment and served as a retrospectively built control group. Scrotal cooling at night by means of a perigenital air stream resulted in a scrotal temperature drop by 0.8 °C (median). A significant increase in sperm concentration and total sperm count was achieved by nocturnal cooling after 8 weeks (p < 0.01; p < 0.05; respectively) and 12 weeks (p < 0.01; p < 0.01; respectively). The improvement of sperm motility and sperm morphology was statistically insignificant. The present study suggests nocturnal scrotal cooling as a therapeutic option to improve semen quality. In a further controlled prospective study the influence on pregnancy rates should be evaluated.
Characterization of a critical region in the hormone binding domain of sperm progesterone receptor
Authors: SACHDEVA GEETANJALI , GADKAR SUSHAMA , SHAH CHIRAG ASHWINKUMAR , KHOLKUTE SANJIVA DATTATRAYA , PURI CHANDER PRAKASH
Published: March 2005

Our previous studies have demonstrated that the ligand binding characteristics of the non‐genomic sperm progesterone receptor (sPR) are different from those of the conventional progesterone receptor (PR). Unlike PR, sPR does not bind efficiently to progesterone antagonists i.e. mifepristone (RU38486), onapristone (ZK98.299) etc. The present study was undertaken to determine the molecular composition of a region that plays a critical role in the interaction of the hormone‐binding domain (HBD) of sPR with progesterone and its antagonists. Detection, cloning and sequencing of the HBD region of sPR revealed its complete sequence homology to the corresponding region in the conventional PR. No nucleotide substitution/mutation/deletion, which could account for the differential antihormone binding, was observed in the HBD of sPR. The critical codon (nucleotide 3216–3218) in all three clones for the HBD of sPR was found to encode for glycine. This ruled out the possibility of steric hindrance because of the placement of amino acids with side chains at a critical position in the HBD, which may interfere in binding of sPR with antiprogestins. It is likely that the post‐transcriptional modifications contribute to the differential binding characteristics of sPR. This warrants future investigations to focus more on the characterization of the mature sPR protein.
A new and simple method to evaluate early membrane changes in frozen–thawed boar spermatozoa
Authors: PEÑA F. J. , SARAVIA F. , JOHANNISSON A. , WALGREN M. , RODRÍGUEZ‐MARTÍNEZ H.
Published: March 2005

Detection of early changes in the sperm plasma membrane during cryopreservation is of utmost importance when designing freezing protocols and has previously been studied in the pig species using annexin‐V detection of phosphatidylserine translocation. In the present study we designed a new assay to detect these changes in boar spermatozoa, based on the slight increase of sperm membrane permeability occurring during the early stages of cryoinjury, using the combination of three fluorescent probes, SNARF‐1, YO‐PRO‐1 and ethidium homodimer. Four ejaculates from five different boars were frozen–thawed and flow cytometrically (FC) evaluated as paired samples. One of the samples was assayed using the annexin‐V/propidium iodide staining and the other sample was evaluated using the new triple staining. Using this combination of probes, four sperm subpopulations were easily detected: viable, with stable membranes (SNARF‐1 positive cells), and three with compromised membranes, one of YO‐PRO‐1+/Eth− cells, one ethidium homodimer+ spermatozoa and, finally spermatozoa stained both with YO‐PRO‐1 and ethidium homodimer (YO‐PRO‐1+/Eth+). The latter three categories corresponded to dead spermatozoa, but with different degree of membrane damage, being YO‐PRO+/Eth− an earlier stage of membrane destabilization, (manifested by an increase in membrane permeability, while still maintaining membrane integrity) than YO‐PRO+/Eth+. A method agreement analysis between both methods was performed revealing good agreement, although the percentage of live cells was 9.44% larger for the triple stain than the annexin‐V assay. The new assay stained all sperm sub‐populations present in the sample, making it especially suitable for both fluorescence microscopy and flow cytometry, facilitating the exclusion of debris and egg‐yolk particles when using FC.
The impact of mitochondrial genetics on male infertility
Authors: JOHN JUSTIN C. ST. , JOKHI ROOBIN P. , BARRATT CHRISTOPHER L. R.
Published: March 2005

Human mitochondrial DNA (mtDNA) encodes 13 of the polypeptides associated with the process of oxidative phosphorylation (OXPHOS), the cells most important ATP generating pathway. Until recently, the effects of mtDNA rearrangements on male fertility have been largely ignored. However, it is becoming increasingly evident that both point mutations and large‐scale deletions may have an impact on sperm motility and morphology. We discuss the implications of these rearrangements in the context of the clinical setting. We further discuss the possible consequences resulting from the transmission of sperm mtDNA deletions to the offspring. The role of nucleo‐cytoplasmic interaction is investigated in the context of nuclear transcription and replication factors that regulate mtDNA transcription and replication.
Ontogenetic profile and thyroid hormone regulation of type‐1 and type‐8 glucose transporters in rat Sertoli cells
Authors: CAROSA ELEONORA , RADICO CARLA , GIANSANTE NADIA , ROSSI SIMONA , D'ADAMO FABIO , DI STASI SAVINO M. , LENZI ANDREA , JANNINI EMMANUELE A.
Published: March 2005

The glucose transporters (GLUTs) gene encode glycoproteins responsible for facilitating transfer of glucose across plasma membrane. In testis, different members of this family are present. In particular the main GLUT mRNA expression within the adult testis is the type 8, while type 1 is more expressed in prepubertal testis. Thyroid hormone, which receptors and function have been characterized in the testis, plays a crucial role in the cellular energetic metabolism. In fact, in the immature Sertoli cells, GLUT1 is up regulated by l‐triiodothyronine (T3). The aim of this paper is to investigate the expression profile of GLUT1 and GLUT8 in the testis during development and in adulthood and analyse the role of T3 on their expression. To analyse the expression of GLUT8 and GLUT1 we performed Northern blot and RT‐PCR experiments in the whole testis and in Sertoli cells from rats of different ages. Treatments in vivo and in vitro with T3 were used to study the effect of thyroid hormones on GLUT1 and GLUT8 expression. The activity of the rat GLUT1 promoter and its regulation by T3 was studied with transient transfections in gonadal and non‐gonadal cell lines and in primary Sertoli cell cultures. GLUT8 is expressed at a low level in the prepubertal testis and Sertoli cells and does not appear to be under T3 control. GLUT1 is the predominant form in immature Sertoli cells. The effect of T3 on its mRNA accumulation was quantified and confirmed by RT‐PCR (control: 0.65 ± 0.17; T3: 1.23 ± 0.04, arbitrary units, p < 0.05). However, transfection experiments showed that T3 does not directly regulate GLUT1 promoter in any cell line tested. This is confirmed by the evidence that, upon extensive analysis, the rat GLUT1 promoter and the first intron sequence do not shows any thyroid responsive elements. Our data demonstrate that GLUT1 and GLUT8 are both expressed in prepubertal testis, but only GLUT1 is regulated by T3. In addition, we found that the effect of T3 cannot be attributed to its action on GLUT1 promoter.
Control of Phytophthora cryptogea in the hydroponic forcing of witloof chicory with the rhamnolipid‐based biosurfactant formulation PRO1
Authors: De Jonghe K. , De Dobbelaere I. , Sarrazyn R. , Höfte M.
Published: March 2005

Rhamnolipids, extracellular metabolites of Pseudomonas aeruginosa with surfactant properties, proved to be very effective in controlling the spread of brown root rot disease caused by Phytophthora cryptogea in the hydroponic forcing system of witloof chicory (Cichorium intybus var. foliosum). The biosurfactant was applied as the product PRO1, a formulation of 25% rhamnolipids in oil. Both an in vitro screening and in vivo experiments in a mini‐hydroponic system demonstrated the ability of PRO1 to control brown root rot. A 25 µg mL−1 rhamnolipids nutrient solution was enough to obtain good control of an artificial infection with a zoospore suspension of P. cryptogea. The biosurfactant PRO1 performed well in a semicommercial system under growers’ conditions. A treatment of 25 µg mL−1 rhamnolipids (100 µg mL−1 PRO1) reduced the disease incidence significantly in two independent experiments. However, PRO1 was not effective when a mycelial suspension was used as inoculum. Rhamnolipids have good potential to limit the spread of P. cryptogea in the hydroponic forcing system of witloof chicory, and can be used as a preventive measure against brown root rot.
Comparison of populations of the wilt pathogen Ceratocystis albifundus in South Africa and Uganda
Authors: Barnes I. , Nakabonge G. , Roux J. , Wingfield B. D. , Wingfield M. J.
Published: March 2005

Ceratocystis albifundus is an important fungal pathogen of Acacia mearnsii trees in South Africa. In a previous study, a high level of gene diversity was demonstrated in a South African population of C. albifundus. This, together with the occurrence of the pathogen on native Protea species and its exclusive occurrence in South Africa, led to the hypothesis that C. albifundus is probably native to that country. More recently, C. albifundus has been reported from A. mearnsii in south‐western Uganda. The aim of this study was to compare the populations of C. albifundus from Uganda and South Africa based on genetic diversity, population structure and possible gene flow. This was achieved using codominant microsatellite markers developed for the closely related species Ceratocystis fimbriata. Available isolates for comparison were from six different areas of South Africa and six jungle stands in Uganda. Eight of the 11 available markers amplified loci in C. albifundus. Gene diversity was higher in the Ugandan population, but genotypic diversity was greater for the South African isolates. There were no common genotypes between the two populations and they shared only 22% of the total alleles. The populations were genetically isolated from each other and highly substructured within. There was no association between isolates collected from the same geographic locations, and gene flow between the two populations was low. Results suggest that C. albifundus was probably not introduced into Uganda from South Africa but rather that an ancestral population, yet to be discovered, is the source of both populations.
Ustilospores of Tilletia ehrhartae, a smut of Ehrharta calycina, are common contaminants of Australian wheat grain, and a potential source of confusion with Tilletia indica, the cause of Karnal bunt of wheat
Authors: Pascoe I. G. , Priest M. J. , Shivas R. G. , Cunnington J. H.
Published: March 2005

Australian wheat consigned for export from Australian ports was surveyed in March 2004 using a national diagnostic protocol for detection and identification of Tilletia indica. No ustilospores of T. indica were detected, confirming previous surveys which have failed to detect T. indica in Australia. However, the survey detected moderate levels of the common smuts Tilletia caries (syn. Tilletia tritici), Tilletia laevis and Urocystis agropyri, and very low levels (average fewer than six ustilospores per 150 g sample) of an unidentified dark, tuberculate‐spored Tilletia in ≈ 60% of samples tested. Comparison with herbarium specimens enabled identification of the majority of the tuberculate ustilospores as Tilletia ehrhartae, a smut fungus known to infect only Ehrharta calycina (perennial veldt grass) and which is common in southern Australia. A smaller number of tuberculate smut ustilospores were identified as Tilletia walkeri, a smut of Lolium spp. recorded in Australia but apparently uncommon. Both T. ehrhartae and T. walkeri bear sufficient resemblance to T. indica for misidentifications to be possible where only a very few ustilospores are seen, although T. ehrhartae ustilospores are always <25 µm in diameter. The frequent presence of ustilospores of both T. ehrhartae and T. walkeri as contaminants of Australian wheat grain exports has significance for diagnosticians testing Australian export wheat, as it demonstrates the potential for tuberculate ustilospores of species other than those covered in existing diagnostic protocols to be misidentified as T. indica. This paper describes T. ehrhartae in detail, and provides criteria for its differentiation from T. indica, T. walkeri and some other species.
Erwinia amylovora populations resistant to oxolinic acid in Israel: prevalence, persistence and fitness
Authors: Kleitman F. , Shtienberg D. , Blachinsky D. , Oppenheim D. , Zilberstaine M. , Dror O. , Manulis S.
Published: March 2005

Oxolinic acid (OA) has been the only bactericide used against fire blight in pear and quince orchards in Israel since 1998. OA‐resistant Erwinia amylovora strains (Ea‐OAR) were detected in several orchards in two restricted areas in the northern Galilee region during 1999–2001. In the following years, resistant strains could not be detected in some of these locations. Documenting the fate of Ea‐OAR strains in commercial orchards at eight sites in northern Israel during 2000/03 revealed that the resistant population appeared irrespective of the number of sprays applied and the severity of the disease. The persistence of the Ea‐OAR populations varied from site to site, ranging from 4 to 20 months; these differences could be attributed to the fire blight management activities of growers. Comparative studies on the fitness of Ea‐OAR and E. amylovora strains sensitive to OA (Ea‐OAS) were conducted in vitro and in planta using two strains of each group. In four of the six comparisons, disease incidence on detached blossoms inoculated with Ea‐OAS was significantly higher than that on blossoms inoculated with Ea‐OAR. In two experiments conducted on 8‐year‐old pear trees grown under netting, the colonization of Ea‐OAS in blossoms, annual shoots and perennial spurs was significantly higher than that of the Ea‐OAR. In two experiments conducted on 2‐year‐old trees grown under netting in an experimental station, the incidence of shoots exhibiting fire blight symptoms and the rate of symptom progress within the branches were significantly higher in trees inoculated with Ea‐OAS than in those inoculated with Ea‐OAR. The results of this study suggest that OA‐resistant E. amylovora strains have lower fitness than wild‐type strains. These findings may have implications for fire blight management.
Distribution and properties of geographically distinct isolates of sugar beet yellowing viruses
Authors: Stevens M. , Patron N. J. , Dolby C. A. , Weekes R. , Hallsworth P. B. , Lemaire O. , Smith H. G.
Published: March 2005

From a total of 261 yellow sugarbeet leaves collected from 10 countries representing three continents, the incidence and distribution of strains of Beet mild yellowing virus (BMYV), Beet chlorosis virus (BChV) and Beet yellows virus (BYV) were analysed using serological and molecular methods. BMYV was found in all countries except Greece, and more frequently in the northern and western areas of Europe, whereas BYV predominated in Turkey, Spain, Greece, the USA and Chile. BChV, originally found in the USA and the UK in 1989, was identified in France, Spain, the Netherlands and Chile. Nine sugar beet poleroviruses, plus a reference isolate of Turnip yellows virus (TuYV, syn. Beet western yellows virus), were further characterized and compared. Isolates obtained from sugar beet infected this species, but not oilseed rape or lettuce; all isolates except one infected Capsella bursa‐pastoris. The coat‐protein sequences of these isolates were highly similar, with the consensus sequence representing 89% of nucleotide residues. Within the coat‐protein gene, two regions were identified that could represent specific epitopes to which monoclonal antibody BYDV‐PAV‐IL‐1 could bind; this antibody is used to distinguish beet poleroviruses in ELISA. Comparison of the sequences at the 5′ end showed that sequence homology existed only between isolates with the same host range. The first sequence data of polerovirus isolates from Chile are presented, showing that the coat protein and the 5′ end of their genomes are highly similar to those of BMYV isolates found in Europe. Chilean polerovirus isolates may have been imported from the northern hemisphere in sugar beet breeding material.
Isolation, identification and molecular characterization of physiological races of apple powdery mildew (Podosphaera leucotricha)
Authors: Urbanietz A. , Dunemann F.
Published: March 2005

The development of apple varieties displaying durable resistance against powdery mildew is one of the major aims in apple breeding programmes worldwide. For a reliable judgment of the resistance of different Malus genotypes, an extended knowledge about the virulence of the pathogen is necessary. To prove the existence of physiological races of Podosphaera leucotricha, 31 monoconidial isolates of the obligate biotrophic fungus representing five locations within Europe have been established and maintained over a period of 3–4 years. The isolates were maintained on in vitro shoots of the highly susceptible apple cv. Gibb's Golden Gage. An AFLP‐based DNA fingerprinting protocol was developed and, using 54 stably reproducible AFLP markers, a dendrogram revealed genetic variability among different isolates of P. leucotricha. Although the molecular characterization of the isolates showed an overall low level of genetic variability, the high phenotypic diversity among European isolates suggest that sexual reproduction may also be involved in the disease cycle of the pathogen in Europe. Phytopathological tests using detached leaves of a collection of 36 Malus genotypes allowed the differentiation of five selected isolates by their virulence patterns. A high level of diversity in terms of virulence was obtained in P. leucotricha. From the present study, based on apple breeding germplasm, cultivars and Malus species, it can be concluded that physiological races of P. leucotricha do indeed exist in Europe.
Effect of temperature on head blight of wheat caused by Fusarium culmorum and F. graminearum
Authors: Brennan J. M. , Egan D. , Cooke B. M. , Doohan F. M.
Published: March 2005

The effect of small temperature differentials (16 vs. 20°C) on the pathogenicity of deoxynivalenol producing single isolates of Fusarium culmorum and F. graminearum and on the fusarium head blight (FHB) response of eight wheat cultivars was examined. Fusarium culmorum inoculation caused greater visual disease symptoms at 20°C than at 16°C, both overall and on an individual cultivar basis (overall AUDPC = 13·5 and 9·6, respectively) (P < 0·05). In contrast, F. graminearum inoculation caused greater overall visual disease symptoms at 16°C than at 20°C, both overall and at the individual cultivar level (overall AUDPC = 12·8 and 10·9, respectively) (P < 0·05). Results showed both F. culmorum and F. graminearum inoculations caused a greater loss in yield at 20°C (54·3 and 46·9% relative 1000‐grain weight, respectively) compared with 16°C (73·3 and 66·9% relative 1000‐grain weight, respectively) (P < 0·05). Fusarium culmorum‐inoculated heads contained similar amounts of fungal DNA at both 16 and 20°C (1·9 and 1·7 ng mg−1 of plant material, respectively) (not significant), while for F. graminearum inoculation, plants contained higher amounts of fungal DNA at 20°C (2·0 and 1·0 ng mg−1 of plant material, respectively) (P < 0·05). Overall, there was a significant negative correlation between AUDPC and percentage relative 1000‐grain weight at both 16 and 20°C (r =−0·693 and −0·794, respectively, P < 0·01).
Effect of formulation on the rhizosphere competence and biocontrol ability of Trichoderma atroviride C52
Authors: McLean K. L. , Swaminathan J. , Frampton C. M. , Hunt J. S. , Ridgway H. J. , Stewart A.
Published: March 2005

The rhizosphere competence of the biological control agent Trichoderma atroviride isolate C52 was studied on onion roots both in the glasshouse and in the field when introduced into soil in a range of formulations. Proliferation of T. atroviride in the rhizosphere was formulation‐dependent. A pellet formulation maintained the fungal concentration at 105 cfu per g soil, whereas solid‐substrate and seed‐coating formulations gave concentrations of 104 and 101 cfu per g soil, respectively. To facilitate rhizosphere‐competence studies, a UP‐PCR band profile generated with primer L45 for isolate C52 was used to enable conclusive identification of T. atroviride C52 when recovered from soil. When isolate C52 was introduced into Sclerotium cepivorum‐infested soil as both pellet and solid‐substrate formulations, there was no statistically significant difference in the disease control between these treatments, but the pellet treatment doubled the percentage of healthy plants compared with the control treatment.
Double‐stranded RNA: distribution and analysis among isolates of Rhizoctonia solani AG‐2 to ‐13
Authors: Bharathan N. , Saso H. , Gudipati L. , Bharathan S. , Whited K. ,   K. Anthony
Published: March 2005

The occurrence, distribution, and genetic relatedness of double‐stranded RNA (dsRNA) components from 36 isolates of Rhizoctonia solani belonging to nine anastomosis groups (AGs) were studied using electrophoretic analysis and RNA–RNA blot hybridization. DsRNA was consistently detected in all 36 isolates. The size of the dsRNA components varied considerably, ranging from 0·74 to 23 kb. Two thirds of isolates possessed different size dsRNA components. Only two of the isolates had small size dsRNA between 0·5 and 1·0 kb. The biotin‐labelled dsRNA probes provided the sensitivity and specificity required to study genetic relationships of dsRNA. As little as 10 pg of ‘hybridizing’ dsRNA could be detected using biotin‐labelled total dsRNA with no detectable nonspecific‐hybridization. Results from several dot‐spot as well as RNA–RNA gel hybridization experiments revealed considerable sequence heterogeneity among dsRNA components within each isolate or isolates from the same AG.
Cultivar preference exhibited by two sympatric and genetically distinct populations of the soybean fungal pathogen Phialophora gregata f.sp. sojae
Authors: Meng X. , Grau C. R. , Chen W.
Published: March 2005

Phialophora gregata f.sp. sojae, a soilborne vascular pathogen causing brown stem rot of soybean, has been divided into A and B populations based on variation in the intergenic spacer region of nuclear rDNA (rDNA marker). The A and B populations correlate with defoliating and nondefoliating pathotypes, respectively. In this study, eight additional polymorphic anonymous marker loci (five inter simple sequence repeat loci and three long‐primer random amplified polymorphic DNA loci) were identified and applied to a total of 189 isolates. Alleles of these eight loci were invariant within, but different between the A and B populations, providing further evidence that the rDNA marker identifies genetically distinct populations. The two populations were sympatric, residing not only in the same field, but also in the same plants under field conditions. Representative strains of the two populations, when used individually in inoculations, infected both resistant cv. Bell and susceptible cv. Sturdy. However, when the same representatives of the two populations were mixed in a 1 : 1 ratio and used as a mixed inoculum in a competitive bioassay, differential cultivar preference was revealed using PCR detection of populations in infected plants. Population A was detected significantly more often (18 out of 24 plants) in the susceptible cv. Sturdy, whereas population B was detected significantly more often (17 out of 24 plants) in the resistant cv. Bell, corroborating earlier field studies. This is the first controlled experiment to demonstrate a differential host preference of P. gregata f.sp. sojae toward different cultivars of the same host species. Unification of terminologies used in P. gregata f.sp. sojae is discussed.
Potential role for salicylic acid in induced resistance of asparagus roots to Fusarium oxysporum f.sp. asparagi
Authors: He C. Y. , Wolyn D. J.
Published: March 2005

Pre‐inoculation of asparagus (Asparagus officinalis) roots with selected nonpathogenic isolates of Fusarium oxysporum (npFo) has previously been shown to induce systemic resistance against infection by F. oxysporum f.sp. asparagi (Foa) through activation of plant‐defence mechanisms. To elucidate the putative npFo‐mediated defence pathways, the effect of salicylic acid (SA) was examined in a split‐root system of asparagus where one half of the seedling root system was drenched with SA and the activation of defence responses was measured subsequently on the remaining roots. SA‐treated plants exhibited enhanced systemic resistance, with a significant reduction in disease severity of the roots inoculated with Foa, compared with untreated plants. SA activated peroxidase and phenylalanine ammonia‐lyase, as well as lignification, upon Foa attack, in a manner similar to that observed with npFo pretreatment. In addition, application of diphenyleneiodonium, an SA biosynthesis inhibitor, led to failure of npFo to induce lignin deposition and systemic resistance. Treatment of fungal spores with SA did not affect germination and growth of either npFo or Foa in in vitro antifungal assays. Production of SA at the site of npFo infection may be involved in the induction of Foa resistance in asparagus roots.
Presence of the Stb6 gene for resistance to septoria tritici blotch (Mycosphaerella graminicola) in cultivars used in wheat‐breeding programmes worldwide
Authors: Chartrain L. , Brading P. A. , Brown J. K. M.
Published: March 2005

Fifty‐two wheat cultivars and breeding lines, most of which have been used in breeding programmes worldwide, were tested for isolate‐specific resistance to Mycosphaerella graminicola isolate IPO323, which interacts with the Stb6 gene of wheat (first identified in cvs Flame and Hereward) via a gene‐for‐gene relationship. Twenty‐three lines were specifically resistant to this isolate. Sixteen resistant lines were crossed with Flame for a test of allelism. All progeny lines were resistant, suggesting that the 16 parental lines had Stb6, a gene allelic to it or a gene closely linked to it. In 14 lines, resistance to IPO323 was controlled by Stb6 only. An exception was Kavkaz‐K4500 L6.A.4., which has two genes for resistance to IPO323, one of which is Stb6. The microsatellite marker Xgwm369 was used to examine genetic diversity in the region of the genome containing Stb6, to which it is closely linked. Eleven alleles of Xgwm369, with amplified fragments of 10 different sizes, as well as apparent nonamplification of this marker in Bulgaria 88, were detected. Through the use of information about lines’ ancestry, combined with Xgwm369 alleles, it was shown that Stb6 entered world wheat‐breeding programmes on a minimum of six occasions, and possibly from as many as 11 sources. The presence of Stb6 in both European and Chinese landraces suggests that this gene has been present in cultivated wheat since the earliest times of agriculture.
Development ofOculimacula yallundaeandO. acuformis(eyespot) on leaf sheaths of winter wheat in the UK in relation to thermal time
Authors: Wan A. M. , Bock C. H. , Fitt B. D. L. , Harvey J. L. , Jenkyn J. F.
Published: March 2005

In a controlled environment (15/10°C) (day/night) container experiment on winter wheat (cv. Avalon), eyespot incidence (percentage of plants affected) and number of leaf sheaths penetrated after 6 weeks increased with inoculum concentration (102−106conidia mL−1) ofOculimacula yallundae(OY) orOculimacula acuformis(OA), but there was no difference between the two species. In an outdoor container experiment, seedlings inoculated with OY 2 weeks after sowing had a greater incidence of eyespot than those inoculated with OA, when assessed 7 weeks after inoculation. Seedlings inoculated with OA at 10 or 20 weeks after sowing developed more severe eyespot by maturity than those inoculated with OY. In an experiment at 15/10°C with seedlings inoculated with OY + OA 2 weeks after sowing, more leaf sheaths were penetrated by OY (3·0 per plant) than OA (2·3 per plant) 6 weeks after inoculation. Field experiments with winter wheat consistently showed leaf sheath production, leaf sheath death, and number of leaf sheaths infected or penetrated by OA or OY were related linearly to thermal time (°C days) after sowing. Depending on cultivar, season and sample, a new leaf sheath was produced in 116–216°C days; a leaf sheath died in 221–350°C days; and infection of a new leaf sheath occurred in 129–389°C days. The mean number of living leaf sheaths infected differed between samples, cultivars and seasons for both OY and OA. Regression analysis of the 1985/86 data suggested that OY progressed more rapidly than OA through the leaf sheaths, and that both the pathogens progressed more rapidly than the rate of leaf sheath death, but more slowly than the rate at which leaf sheaths were produced. It also suggested that OA progressed more slowly than the rate at which leaf sheaths died in 1987/88, but OY did not.
Evaluation of foliar resistance in potato to Phytophthora infestans based on an international field trial network
Authors: Hansen J. G. , Koppel M. , Valskyte A. , Turka I. , Kapsa J.
Published: March 2005

During the period 2000–03, local potato cultivars in Estonia, Latvia, Lithuania, Poland and Denmark were tested for foliar resistance to Phytophthora infestans (late blight) in an international field trial network. Four standard cultivars were included in the trials: Sava, Oleva, Danva and Kuras. Primary disease‐assessment data were entered into a common database, and parameters from the disease progress curves were calculated and made available on interactive web pages. A regression model, using relative area under disease progress curve (RAUDPC) values for cv. Oleva as a reference, was developed for the estimation of 1–9 scale values, where 1 = most susceptible. Standard deviations for the estimated 1–9 scale values and a nonparametric rank stability analysis of RAUDPC were used to evaluate the stability of resistance of the cultivars. Overall, the results showed stability of resistance for cvs Sava, Oleva and Danva, but not for Kuras. Use of the Internet‐based Web‐Blight service in this study facilitated comparison of results among countries for the level and stability of resistance. The estimated 1–9 scale values were similar to, or slightly lower than, those from official cultivar lists or from the European Cultivated Potato Database, especially for the more resistant cultivars. Possible reasons for discrepancies from different sources and locations are discussed. It is concluded that RAUPDC and the derived 1–9 scale values are useful for ranking cultivars for resistance to P. infestans, but this information is not detailed enough for use in a decision support system for late blight control.
Meconium Aspiration Syndrome Induces Complement‐Associated Systemic Inflammatory Response in Newborn Piglets
Authors: Castellheim A. , Lindenskov P. H. H. , Pharo A. , Aamodt G. , Saugstad O. D. , Mollnes T. E.
Published: March 2005

The pathophysiology of meconium aspiration syndrome (MAS) is complex. We recently showed that meconium is a potent activator of complement. In the present study, we investigated whether the complement activation occurring in experimental MAS is associated with a systemic inflammatory response as judged by granulocyte activation and cytokine and chemokine release. MAS was induced by the instillation of meconium into the lungs of newborn piglets (n = 8). Control animals (n = 5) received saline under otherwise identical conditions. Haemodynamic and lung dynamic data were recorded. Complement activation, revealed by the terminal sC5b‐9 complex (TCC), and cytokines [interleukin (IL)‐6 and IL‐8] were measured in plasma samples by enzyme immunoassays. The expression of CD18, CD11b and oxidative burst in granulocytes was measured in whole blood by flow cytometry. Plasma TCC increased rapidly in the MAS animals in contrast with controls (P < 0.0005). The TCC concentration correlated closely with oxygenation index (r = 0.48, P < 0.0005) and ventilation index (r = 0.57, P < 0.0005) and inversely with lung compliance (r = −0.63, P < 0.0005). IL‐6 and IL‐8 increased in MAS animals compared with the controls (P = 0.002 and P < 0.001, respectively). Granulocyte oxidative burst declined significantly in the MAS animals compared with the controls (P < 0.02). TCC correlated significantly with IL‐6 (r = 0.64, P < 0.0005) and IL‐8 (r = 0.32; P = 0.03) and inversely with oxidative burst (r = −0.37; P = 0.02). A systemic inflammatory response associated with complement activation is seen in experimental MAS. This reaction may contribute to the pathogenesis of MAS.
Increased Frequency of C4B*Q0 Alleles in Patients with Henoch–Schönlein Purpura
Authors: Stefansson Thors V. , Kolka R. , Sigurdardottir S. L. , Edvardsson V. O. , Arason G. , Haraldsson A.
Published: March 2005

Henoch–Schönlein purpura (HSP) is a vasculitis of unknown aetiology, possibly involving immune complexes. The complement system is essential for the clearance of immune complexes. Our aim was to explore the hypothesis that patients with HSP have abnormal complements, contributing to the development of the disease. The study included 56 patients diagnosed with HSP at the Children's Hospital, Iceland between 1984 and 2000, and 98 blood donors as controls. Serum levels of immunoglobulin A, C4A, C4B and mannan‐binding lectin were measured and compared between the two groups. C4 null alleles were significantly more common in HSP patients than in controls (P = 0.018) and were carried by 66.1% of the patients compared with 41.2% of the controls. This difference was due to an increased frequency of C4B*Q0 allele in the HSP group (0.25 versus 0.11 in the control group; P = 0.002). The fact that the majority of our patients carried a C4 null allele indicates that children with C4 deficiencies may have an increased risk of developing HSP. This may reflect inadequate complement activity and possibly present an opportunity to identify patients at risk of developing serious morbidity associated with HSP.
The Inhibitory Effect of Altered Collagen II Peptide on HLA‐DRB1‐Restricted T‐Cell Activation
Authors: Cheng Y. J. , Zhou Q. , Li Z. G.
Published: March 2005

It has been known that rheumatoid arthritis (RA)‐associated antigenic peptides CII263–272 are coupled with human leucocyte antigen (HLA)‐DRB1 and recognized by T‐cell receptor (TCR), which in turn induced T‐cell proliferation and pathogenesis of RA. Non‐T‐cell‐stimulating type II collagen (CII) peptides might be generated by removing the amino acids responsible for TCR contact and keeping the HLA‐DR‐binding residues intact. In this study, a panel of altered CII peptides (APs) with consecutive or single substitutions of the TCR‐contacting residues were synthesized. Through peptide binding and T‐cell activation assays, we demonstrated that altered CII263–272 peptides with substitution of the TCR‐contacting residues did not or barely induced T‐cell activation; one of the best non‐T‐cell‐stimulating peptide AP268–270 inhibited the binding of wild‐type CII263–272 to HLA‐DR1 and T‐cell activation triggered by wild‐type CII263–272 and HA306–318 in a dose–response manner. These data suggest that removal of the TCR‐contacting residues of CII263–272 leads to HLA‐DRB1 binding and low T‐cell‐stimulating peptides, which could potentially inhibit the T‐cell response induced by HLA‐DRB1‐binding antigenic peptides.
Mycobacterium avium ssp. paratuberculosis Recombinant Heat Shock Protein 70 Interaction with Different Bovine Antigen‐Presenting Cells
Authors: Langelaar M. F. M. , Hope J. C. , Rutten V. P. M. G. , Noordhuizen J. P. T. M. , Van Eden W. , Koets A. P.
Published: March 2005

Heat shock proteins (Hsp) can deliver antigen into the major histocompatibility complex class I presentation pathway of antigen‐presenting cells (APC), a process called cross priming, thus stimulating antigen‐specific CD8+ T‐cell reactions. Hsp were shown to elicit proinflammatory responses in APC. Both processes require interaction of Hsp with APC via specific receptors. This study describes the interaction of recombinant Hsp70 (rHsp70) of Mycobacterium avium subspecies paratuberculosis with bovine peripheral blood mononuclear cells that was restricted to CD14+ cells. Characterized monocyte‐derived macrophages, monocyte‐derived dendritic cells (DC) and BoMac, an immortalized bovine macrophage cell line, were used to investigate the interaction of rHsp70 with different bovine APC. Saturation of immature DC with high concentrations of rHsp70 is demonstrated, and it was found that interaction of rHsp70 with DC was related to the maturation stage of the DC. Involvement of CD91 as a cellular receptor for rHsp70 was demonstrated; however, competition studies with immature DC demonstrated that other receptors exist on bovine APC. These data suggest that rHsp70‐based vaccines may be useful for the successful immunization of cattle.
Inhibition by Artocarpus tonkinensis of the Development of Collagen‐Induced Arthritis in Rats
Authors: Ngoc D. D. T. , Catrina A. I. , Lundberg K. , Harris H. E. , Ha N. T. , Anh P. T. , Larsson P.
Published: March 2005

Extracts of the leaves and roots from the tree Artocarpus tonkinensis A Cheval (family Moraceae) are used in traditional Vietnamese medicine in order to treat backache as well as rheumatic joint diseases. We prepared an ethyl acetate (EtOAc) extract from this plant and tested its anti‐inflammatory properties in an experimental arthritis model, collagen‐induced arthritis (CIA). CIA was induced in Dark Agouti rats by means of immunization with collagen type II (CII) emulsified in incomplete Freund's adjuvant. Starting at the day of immunization, the rats were treated daily with intraperitoneal injections of Artocarpus extract. Arthritis progression was measured by means of clinical scoring of paws and anti‐CII antibody titres were measured by means of ELISA. In vitro, lymph node (LN) cell cultures were treated with Artocarpus extract and the apoptosis‐inducing effect was determined with FACS staining by using annexin V and propidium iodide as well as the TUNEL method. Treatment of the rats with Artocarpus extract decreased arthritis incidence and severity and delayed disease onset. When treatment was started after the onset of arthritis, a tendency towards arthritis amelioration was observed. In vitro, Artocarpus extract acted as a T‐cell modulator, inhibiting mitogen‐induced T‐cell proliferation and inducing apoptosis of activated LN‐derived lymphocytes. Thus, we have demonstrated that an EtOAc extract of Artocarpus, a plant traditionally used in Vietnamese folk medicine for treating arthritic conditions, has beneficial effects in an experimental arthritis model. This effect is likely to be T cell‐dependent and mediated through apoptosis induction in activated cells.
T‐Cell Receptor Vβ Repertoire after Myeloablative and Reduced Intensity Conditioning Allogeneic Haematopoietic Stem Cell Transplantation
Authors: Hentschke P. , Omazic B. , Mattsson J. , Näsman‐Björk I. , Lundkvist I. , Gigliotti D. , Barkholt L. , Ringdén O. , Remberger M.
Published: March 2005

T cells play an important role in the adaptive immune system. After haematopoietic stem cell transplantation (HSCT), T‐cell function is impaired. This is reflected by the emergence of opportunistic infections, infections that are often difficult to treat because of the patient's insufficient immune function. T‐cell receptor reconstitution was studied using CDR3 spectratyping to analyze the diversity of the T‐cell repertoire at 3, 6 and 12 months after myeloablative and reduced intensity conditioning (RIC) HSCT in 23 patients. Immune function in vitro was tested by lymphocyte stimulation at 3, 6 and 12 months after HSCT. Lower diversity in the CDR3 repertoire was demonstrated in CD4+ cells after RIC HSCT at 3 and 6 months and in CD8+ cells at 3 months compared with healthy donors. After myeloablative HSCT, lower diversity was seen at 3, 6 and 12 months in CD4+ cells and at 6 and 12 months in CD8+ cells after HSCT. Acute and chronic graft‐versus‐host‐disease (GVHD) did not affect diversity. Responses to phytohaemagglutinin (PHA), Concanavalin A (Con A) and Staphylococcus aureus protein A were significantly lower compared with healthy donors during the first 6 months after RIC HSCT. After myeloablative HSCT, lymphocyte response to Con A was significantly lower at 3 months compared with healthy donors. Decreased responses to cytomegalovirus and varicella zoster virus antigens were seen in patients suffering from acute GVHD grade II or chronic GVHD. The T‐cell repertoire is skewed under the first year after HSCT, and immune reconstitution after HSCT with myeloablative and RIC conditioning seems to be comparable. GVHD, infections and age are more important for immune reconstitution than type of conditioning.
Current status of ABO‐incompatible kidney transplantation in children
Authors: Shishido S. , Hasegawa A.
Published: March 2005

Abstract:  This article reviews the current status of ABO‐incompatible kidney transplantation in the pediatric population. ABO blood type incompatibility between a donor and recipient was generally considered a contraindication to kidney transplantation because of the associated high risk for hyperacute rejection. However, due to a severe shortage of suitable cadaveric allografts, much effort has been made over the last decade to investigate whether successful and effective kidney transplantation is possible across the ABO blood group barrier. At present, ABO‐incompatible kidney transplantation has been shown to be a valid alternative even for children with end stage renal disease. In this review, we will discuss protocols available for successfully performing ABO‐incompatible kidney transplantation in children: (1) pre‐transplant extracorporeal immunomodulation with removal of pre‐existing anti‐A and/or anti‐B antibodies; (2) immunosuppressive therapy and anti‐rejection therapy; (3) splenectomy and the associated infectious complication in asplenic children. Also, we will speculate regarding the mechanisms underlying accommodation following transplantation.
An unusual case of pyelonephritis in a pediatric renal transplant recipient
Authors: Al‐Makadma Abdulkarim S. , Al‐Akash Samhar I.
Published: March 2005

Abstract:  Emphysematous (Gas‐forming) pyelonephritis (EPN) is a rare and life‐threatening infection of the renal parenchyma with or without involvement of the peri‐renal tissues. Diagnosis of this condition is usually made on clinical and radiological grounds. Risk factors for this condition include diabetes mellitus, urinary tract obstruction, renal anomalies such as polycystic kidneys, stones, old age and others. Review of the literature did not reveal any reports of (EPN) in the pediatric age group especially in transplanted patients. We report the first case of EPN in a child, a 12‐yr‐old boy with a renal transplant, and discuss the various diagnostic and management issues of this particular case and EPN in general.
Open lung biopsy in pediatric patients with respiratory failure after abdominal transplantation
Authors: Vendryes Christopher , McLaughlin Gwenn E. , Romaguera Rita L. , Sola Juan , Kato Tomoaki
Published: March 2005

Abstract:  To understand the utility of open lung biopsy (OLB) in the evaluation of respiratory failure in pediatric abdominal organ transplant we reviewed the records of nine children in this patient population who underwent an OLB. Eight of nine patients had undergone a previous non‐diagnostic bronchoalveolar lavage. Biopsies were performed at the bedside in the pediatric intensive care unit and tissue was processed by the Department of Pathology with special stains for infectious agents. There were no significant complications of OLB. A specific treatable etiology was identified in four patients (respiratory syncytial virus, adenovirus, graft‐vs.‐host disease and post‐transplant lymphoproliferative disease), leading to a change in therapy and survival in two. Overall survival was 44%. Given the low morbidity, OLB as performed in this study appears appropriate in this patient population.
Carcinoma of donor origin after liver‐intestine transplantation in a child
Authors: Neto Joao Seda , Macedo Camila , Jaffe Ronald , Mazariegos George V. , Sindhi Rakesh , Bond Geoffrey J. , Abu‐Elmagd Kareem , Reyes Jorge
Published: March 2005

Abstract:  Tumor‐related complications after intestinal transplantation in children have been principally EBV driven post‐transplant disorders. We describe the clinical course of a child, with a diagnosis of microvillus inclusion disease who received a liver and intestine allograft at the age of 9 months. His postoperative course was significant for multiple episodes of acute intestinal allograft rejection and eventually the development of post‐transplant lymphoproliferative disorder (PTLD), which resolved. At 8 yr post‐transplant he presented with masses in the intestine allograft mesentery and in the right lobe of the allograft liver, biopsy of which revealed a relatively undifferentiated tumor, suggestive of a carcinoma. In situ hybridization for X and Y chromosomes, revealed his tumor to be of donor origin. Treatment included debulking of the mesenteric mass with segmental enterectomy of the intestinal allograft, and stopping his immunosuppression for a period of 4 months; this resulted in complete resolution of his malignancy. Immunosuppression with tacrolimus and steroids was restarted because of intestinal allograft rejection; he died suddenly of unknown causes at 17 months post‐diagnosis of carcinoma.

The severely immunosuppressed state produced in this patient allowed for the development of an unusual donor derived carcinoma, which resolved spontaneously with withdrawal of immunosuppression. The mechanism of such regression of tumor may be related to restitution of immunologic competence, but is yet to be determined.