bluuebunny/crossref_metadata_2025 huggingface dataset:
Biology and Molecular Characterization of Cucurbit leaf crumple virus, an Emergent Cucurbit-Infecting Begomovirus in the Imperial Valley of California
Authors: Hagen C. , Rojas M. R. , Sudarshana M. R. , Xoconostle-Cazares B. , Natwick E. T. , Turini T. A. , Gilbertson R. L.
Published: April 2008

Cucurbit leaf crumple virus (CuLCrV) is an emergent and potentially economically important bipartite begomovirus first identified in volunteer watermelon plants in the Imperial Valley of southern California in 1998. Field surveys indicated that CuLCrV has become established in the Imperial Valley; and field plot studies revealed that CuLCrV primarily infects cucurbits, including cantaloupe, squash, and watermelon. Full-length DNA-A and DNA-B clones of an Imperial Valley isolate of CuLCrV were obtained by polymerase chain reaction (PCR) with overlapping primers. These clones were infectious in various cucurbits and common bean (cv. Topcrop); symptoms included stunted growth and leaf crumple, curl, and chlorosis. CuLCrV was not sap-transmissible, and immunolocalization and DNA in situ hybridization studies revealed that it is phloem-limited. A CuLCrV agroinoculation system was generated, and host range studies revealed differential susceptibility in cucurbits, with squash, watermelon, cantaloupe, and honeydew melon being most to least susceptible, respectively. Germplasm screening studies identified a number of resistant cantaloupe and honeydew melon cultivars. The genome organization of this CuLCrV isolate (CuLCrV-CA) is similar to other bipartite begomoviruses, and phylogenetic analysis placed CuLCrV in the Squash leaf curl virus (SLCV) cluster of New World bipartite begomoviruses. A CuLCrV-specific PCR test was developed which allows for differentiation from other begomoviruses, including SLCV.
Colonization of the Residues of Diverse Plant Species by Gibberella zeae and Their Contribution to Fusarium Head Blight Inoculum
Authors: Pereyra S. A. , Dill-Macky R.
Published: April 2008

The presence of Fusarium spp. was examined in the residues of wheat, barley, corn, sunflower, pasture, and gramineous weed species common in wheat and barley cropping systems collected from no-tillage and reduced-tillage plots from February 2001 to March 2003 in Uruguay. Gibberella zeae was recovered from residues of wheat, barley, corn, sunflower, fescue, and the gramineous weeds Digitaria sanguinalis, Setaria spp., Lolium multiflorum, and Cynodon dactylon, except from birdsfoot trefoil or white clover. Of the Fusarium spp. obtained, G. zeae was the most frequently recovered from wheat and barley residues, while other species were more common in other crops. G. zeae declined over time in all residues examined. Wheat and barley residues produced more ascospores of G. zeae than corn or other gramineous residues. Sunflower residue did not support ascospore production, indicating that it probably did not contribute to primary inoculum. Wheat and barley residues supported G. zeae colonization longer in no-till than in reduced-tillage production systems and, thus, may represent major contributors to Fusarium head blight (FHB) inoculum in Uruguay. The presence of G. zeae in the gramineous components of pastures, weed species, and sunflower should be considered when implementing control strategies for FHB.
First Report of Tomato chlorotic spot virus Infecting Spilanthes oleracea in Brazil
Authors: Krause-Sakate R. , Nozaki D. N. , Rosa R. A. , Suzuki G. S. , Pavan M. A.
Published: April 2008

Spilanthes oleracea L., popularly known as toothache plant, belongs to the family Asteraceae and is a South American native plant. Fresh leaves can be eaten for their medicinal properties or used by the cosmetics industry for their spilol contents. Plants showing leaf deformation that were collected in a field in São Paulo State, Brazil in March 2005 were suspected to be infected by a virus. Electron microscopy of leaf dip preparations of symptomatic plants revealed pleiomorphic particles typical of tospoviruses. Extracts from these plants prepared with 0.01 M sodium phosphate buffer, pH 7.0, containing 1% sodium sulfite were mechanically inoculated to indicator plants. Chenopodium amaranticolor and Gomphrena globosa were symptomless. Necrotic local lesions were observed on C. quinoa. Necrotic local lesions followed by a systemic necrosis that caused the death of the plants were observed on Datura stramonium, Nicotiana glutinosa, and N. tabacum ‘TNN’ and ‘Turkish’. Concentric rings followed by systemic necrosis and plant death were induced on N. rustica, N. tabacum ‘Havana 425’, N. clevelandii, Physalis floridana, Capsicum annum ‘Magda’, and Solanum lycopersicum ‘Santa Clara’. Total RNA was extracted (1) from infected S. oleracea and N. rustica plants for reverse transcription-PCR amplification with tospovirus specific primers BR60 (5′ CCCGGATCCTGCAGAGCAATTGTGTCA 3′) and BR65 (5′ ATCAAGCCTTCTGAAAGTCAT 3′) (2), which amplified an approximate 440-bp fragment covering part of the nucleocapsid protein gene. This fragment was sequenced (EMBL Accession No. AM887766) and showed 99% nt sequence identity with Tomato chlorotic spot virus (TCSV) (GenBank Accession No. AF521102), a tospovirus species (3). To our knowledge, this is the first report of a tospovirus infecting S. oleracea in Brazil and indicates that this plant might constitute a reservoir of TCSV or other tospoviruses that could also infect tomato and pepper plants.

References: (1) Y. D. Bertheau et al. DNA amplification by polymerase chain reaction (PCR) 1998 in: Methods for the Detection and Quantification of Erwinia carotovora subsp. atroseptica on Potatoes. M. C. N. Perombelon and J. M. van der Wolf, eds. Scott. Crop Res. Inst. Occas. Publ. Dundee, Scotland, 1998. (2) M. Eiras et al. Fitopatol. Bras. 26:170, 2001. (3) F. Lovato et al. Virus Genes 29:321, 2004.
First Report of Pestalotiopsis virgatula Causing Pestalotiopsis Fruit Rot on Rambutan in Hawaii
Authors: Keith L. M.
Published: April 2008

Rambutan (Nephelium lappaceum Linn.) is a tropical, exotic fruit that has a rapidly expanding niche market in Hawaii. Diseased rambutan fruit was commonly observed in orchards in the Hilo and Kona districts of Hawaii Island during 2006. In surveys conducted in January, symptoms appeared as dark brown-to-black spots on mature fruit and blackened areas at the base of spinterns (hair-like projections) of mature and immature fruits. Pieces of infected fruit (cv. R167) were surface sterilized for 2 min in 0.5% NaOCl, plated on potato dextrose agar, and incubated at 24 ± 1°C for 7 days. The fungus growing on PDA was pale buff with sparse, aerial mycelium and acervuli containing black, slimy spore masses. All isolates had five-celled conidia. Apical and basal cells were hyaline, while the three median cells were olivaceous; the upper two were slightly darker than the lower one. Conidia (n = 40) were 20.3 ± 0.1 × 6.8 ± 0.1 μm. There were typically three apical appendages averaging 16.8 ± 0.2 μm long. The average basal appendage was 3.8 ± 0.1 μm long. The fungus was initially identified as Pestalotiopsis virgatula (Kleb.) Stey. on the basis of conidial and cultural characteristics (3). The identification was confirmed by molecular analysis of the 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA amplified from DNA extracted from single-spore cultures with the ITS1/ITS4 primers (1,4) and sequenced (GenBank Accession No. EU047943). To confirm pathogenicity, agar pieces, 3 mm in diameter, from 7-day old cultures were used as inoculum. Five mature fruit from rambutan cv. R134 were rinsed with tap water, surface sterilized with 0.5% NaOCl for 2 min, wounded with a needle head, inoculated in the laboratory, and maintained in a moist chamber for 7 days. Lesions resembling symptoms that occurred in the field were observed on fruit after 7 days. No symptoms were observed on fruit inoculated with agar media. The fungus reisolated from diseased fruit was identical to the original isolates, confirming Koch's postulates. The disease appears to be widespread in Hawaii. Preharvest symptoms may have the potential to affect postharvest fruit quality if fruits are not stored at the proper conditions. Pestalotiopsis spp. have been reported on rambutan in Malaysia, Brunei, and Australia (2). To my knowledge, this is the first report of P. virgatula causing fruit spots on rambutan in Hawaii.

References: (1) G. Caetano-Annolles et al. Curr. Genet. 39:346, 2001. (2) D. F. Farr et al. Fungal Databases. Systematic Botany and Mycology Laboratory. On-line publication. ARS, USDA, 2007. (3) E. F. Guba. Monograph of Pestalotia and Monochaetia. Harvard University Press, Cambridge, MA, 1961. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. 1990.
First Report of Powdery Mildew Elicited by Podosphaera fusca (Synonym Podosphaera xanthii) on Euryops pectinatus in Spain
Authors: Fernández-Ortuño D. , Torés J. A. , Vicente A. de , Pérez-García A.
Published: April 2008

Euryops pectinatus Cass. is an evergreen shrub that is planted extensively in Spain for landscape use. In 2007, powdery mildew outbreaks were observed on E. pectinatus in several nurseries located in the Axarquia area (Malaga, southern Spain). Fungal growth appeared as typical, white, powdery mildew colonies that were restricted to upper leaf surfaces. Initially, individual colonies were small and nearly circular, but later enlarged and coalesced to cover the whole leaf surface. With progress of the disease, all green parts (leaves, stems, and petioles) were covered with a white mycelium. Newly developed leaves especially became rapidly infected. Diseased leaves ultimately dried up and senesced, making nursery plants aesthetically unattractive and unsaleable. Conidiophores were erect, had crenate edge lines, cylindrical foot cells that measured 37.5 to 45.0 × 10.0 to 12.5 μm, and were followed by one to three shorter cells. Conidia were hyaline, ellipsoid to ovoid, borne in chains, and measured 27.5 to 35.0 × 12.5 to 17.5 μm. Conidial length-to-width ratios ranged from 1.6 to 2.4. Conidia possessed conspicuous fibrosin bodies and from their sides produced short germ tubes. No chasmothecia were found. The nuclear rDNA internal transcribed spacer (ITS) regions were amplified by PCR and sequenced (GenBank Accession No. EU424056). On the basis of morphological characteristics of the imperfect state and ITS sequence data, this powdery mildew was identified as Podosphaera fusca (Fr.) U. Braun & N. Shishkoff (1), this isolate belonging to ITS haplotype 15 (group III) (3); this group is considered a separate species, P. xanthii (Castagne) U. Braun & N. Shishkoff by some authors (2). Pathogenicity was confirmed by gently pressing diseased leaves onto leaves of healthy E. pectinatus plants. Plants were incubated in a growth chamber at 25°C, and after 14 days, powdery mildew colonies developed. A similar disease of E. pectinatus was observed in 1999 in California (4). P. fusca parasitizes a large number of asteraceous species including field marigold (Calendula arvensis) and fleabane (Erigeron sp.) weeds, as previously reported in the same area, and ornamentals such as Calendula officinalis, Chrysanthemum spp., and Gerbera spp., which are also grown in the same nurseries and frequently attacked by powdery mildew.

References: (1) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000. (2) U. Braun et al. Schlechtendalia 7:45, 2001. (3) T. Hirata et al. Can. J. Bot. 78:1521, 2000. (4) G. S. Saenz et al. Plant Dis. 84:1048, 2000.
Functional Analyses of Laccase Genes from Fusarium oxysporum
Authors: Cañero D. Cordoba , Roncero M. I. G.
Published: April 2008

Six laccase genes, lcc1, lcc2, lcc3, lcc4, lcc5, and lcc9, of the vascular wilt fungus Fusarium oxysporum were isolated and characterized. All genes have the characteristic conserved domains for copper binding of phenol oxidase enzymes. Targeted inactivation of lcc1, lcc3, and lcc5 resulted in a significant decrease of extracellular laccase activity. Reverse transcription-polymerase chain reaction showed that lcc1, lcc2, and lcc9 were constitutively expressed in culture, whereas lcc3 and lcc5 appeared down and up-regulated, respectively, by PacC. Oxidative stress conditions and phenolic compounds altered the growth rate of the Δlcc3 mutant compared with the wild-type. lcc1, lcc3, and lcc9 were expressed in roots and stems during the infection process. However, inactivation of lcc1, lcc3, and lcc5 had no detectable effects on virulence on tomato plants.
Oidium neolycopersici: Intraspecific Variability Inferred from Amplified Fragment Length Polymorphism Analysis and Relationship with Closely Related Powdery Mildew Fungi Infecting Various Plant Species
Authors: Jankovics T. , Bai Y. , Kovács G. M. , Bardin M. , Nicot P. C. , Toyoda H. , Matsuda Y. , Niks R. E. , Kiss L.
Published: April 2008

Previous works indicated a considerable variation in the pathogenicity, virulence, and host range of Oidium neolycopersici isolates causing tomato powdery mildew epidemics in many parts of the world. In this study, rDNA internal transcribed spacer (ITS) sequences, and amplified fragment length polymorphism (AFLP) patterns were analyzed in 17 O. neolycopersici samples collected in Europe, North America, and Japan, including those which overcame some of the tomato major resistance genes. The ITS sequences were identical in all 10 samples tested and were also identical to ITS sequences of eight previously studied O. neolycopersici specimens. The AFLP analysis revealed a high genetic diversity in O. neolycopersici and indicated that all 17 samples represented different genotypes. This might suggest the existence of either a yet unrevealed sexual reproduction or other genetic mechanisms that maintain a high genetic variability in O. neolycopersici. No clear correlation was found between the virulence and the AFLP patterns of the O. neolycopersici isolates studied. The relationship between O. neolycopersici and powdery mildew anamorphs infecting Aquilegia vulgaris, Chelidonium majus, Passiflora caerulea, and Sedum alboroseum was also investigated. These anamorphs are morphologically indistinguishable from and phylogenetically closely related to O. neolycopersici. The cross-inoculation tests and the analyses of ITS sequences and AFLP patterns jointly indicated that the powdery mildew anamorphs collected from the above mentioned plant species all represent distinct, but closely related species according to the phylogenetic species recognition. All these species were pathogenic only to their original host plant species, except O. neolycopersici which infected S. alboroseum, tobacco, petunia, and Arabidopsis thaliana, in addition to tomato, in cross-inoculation tests. This is the first genome-wide study that investigates the relationships among powdery mildews that are closely related based on ITS sequences and morphology. The results indicate that morphologically indistinguishable powdery mildews that differed in only one to five single nucleotide positions in their ITS region are to be considered as different taxa with distinct host ranges.
Specificity of Incomplete Resistance to Mycosphaerella graminicola in Wheat
Authors: Krenz J. E. , Sackett K. E. , Mundt C. C.
Published: April 2008

We examined interactions between wheat (Triticum aestivum) and Mycosphaerella graminicola, causal agent of Septoria tritici blotch, to determine whether specific interactions occur between host and pathogen genotypes that could be involved in eroding resistance. The moderate resistance of the wheat cultivar Madsen has eroded significantly in the Willamette Valley of Oregon since its release in 1990. Foote is a replacement cultivar expressing moderate resistance and was released in 2000. Isolates of M. graminicola were collected from Foote and Madsen in 2004 and 2005 and tested on each cultivar in growth chamber and greenhouse experiments. There was a significant (P ≤ 0.002) cultivar by isolate source interaction in all experiments when the isolates were tested individually and also when they were tested as bulks of isolates collected from the same cultivar. Though the resistance of Foote is still very effective in the field, isolates sampled from that cultivar at the end of the season in both 2004 and 2005 were of high virulence on Foote, while those collected from Madsen usually were of low virulence on Foote. Foote demonstrated qualitative reactions more typical of a major resistance gene that provides incomplete resistance. Madsen showed a more continuous variation in reaction to M. graminicola isolates. The mean of the isolates collected from Madsen caused significantly (P ≤ 0.05) more disease on Madsen than on Foote for the individual isolates collected in 2005, but not for those collected in 2004. Bulk populations collected from Madsen did not cause significantly more disease on Madsen than did isolates collected from Foote in either 2004 or 2005.
Field Management Effects on Damping-Off and Early Season Vigor of Crops in a Transitional Organic Cropping System
Authors: Baysal Fulya , Benitez Maria-Soledad , Kleinhenz Matthew D. , Miller Sally A. , Gardener Brian B. McSpadden
Published: April 2008

Transitioning farmland to certified organic vegetable production can take many paths, each varying in their costs and benefits. Here, the effects of four organic transition strategies (i.e., tilled fallowing, mixed-species hay, low-intensity vegetables, and intensive vegetable production under high tunnels), each with and without annual compost applications for 3 years prior to assessment, were characterized. Although transition cropping strategies differed in soil chemistry (P < 0.05), the magnitude of the changes typically were marginal and pairwise comparisons were rarely significant. In contrast, the compost amendment had a much greater impact on soil chemistry regardless of cropping strategy. For example, percent C and total P increased by 2- to 5-fold and K increased from 6- to 12-fold. Under controlled conditions, damping-off of both edamame soybean (cv. Sayamusume) and tomato (cv. Tiny Tim) was reduced from 2 to 30% in soils from the mixed-hay transition. In the field, damping-off of both crops was also significantly lower in plots previously cropped to hay (P < 0.05). Although not always significant (P < 0.05), this pattern of suppression was observed in all four of the soybean experiments and three of the four tomato experiments independent of compost application. The compost amendments alone did not consistently suppress damping-off. However, plant height, fresh weight, and leaf area index of the surviving seedlings of both crops were greater in the compost-amended soils regardless of the transitional cropping treatment used (P < 0.05 for most comparisons). These data indicate that mixed-hay cropping during the transition periods can enhance soil suppressiveness to damping-off. In addition, although compost amendments applied during transition can improve crop vigor by significantly enhancing soil fertility, their effects on soilborne diseases are not yet predictable when transitioning to certified organic production.
Ultrastructural Study on Acibenzolar-S-Methyl-Induced Scab Resistance in Epidermal Pectin Layers of Japanese Pear Leaves
Authors: Jiang S. , Park P. , Ishii H.
Published: April 2008

The infection behavior of Japanese pear scab pathogen Venturia nashicola race 1 was studied ultrastructurally in acibenzolar-S-methyl (ASM)-pretreated susceptible Japanese pear (cv. Kousui) leaves to determine the mechanism of ASM-induced scab resistance. On ASM-pretreated leaf surfaces, the infection behavior (conidial germination and appressorial formation) was similar to that on distilled water (DW)-pretreated leaves prior to cuticle penetration by the pathogen. However, after penetration, differentiated behavior was found in epidermal pectin layers and middle lamellae of the ASM-pretreated leaves. Subcuticular hyphae in epidermal pectin layers and middle lamellae of ASM-pretreated pear leaves were observed at lower frequency than in DW-treated leaves. The results indicated that fungal growth was suppressed in ASM-pretreated pear leaves. In the pectin layers of ASM- and DW-pretreated leaves, some hyphae showed morphological modifications, which were used as criteria to judge collapse of hyphal cells, including plasmolysis, necrotic cytoplasm, and cell wall destruction. More hyphae had collapsed in ASM-pretreated leaves than in DW-treated ones. In addition, the cell walls of collapsed hyphae broke into numerous fibrous and amorphous pieces, suggesting that ASM-induced scab resistance might be associated with cell-wall-degrading enzymes from pear plants. In addition, results from morphometrical analysis suggested that the activity or production of pectin-degrading enzyme from hyphae were inhibited by ASM application when compared with DW treatment.
In Planta Distribution of ‘Candidatus Liberibacter asiaticus’ as Revealed by Polymerase Chain Reaction (PCR) and Real-Time PCR
Authors: Tatineni Satyanarayana , Sagaram Uma Shankar , Gowda Siddarame , Robertson Cecile J. , Dawson William O. , Iwanami Toru , Wang Nian
Published: April 2008

Huanglongbing (HLB) is one of the most devastating diseases of citrus worldwide, and is caused by a phloem-limited fastidious prokaryotic α-proteobacterium that is yet to be cultured. In this study, a combination of traditional polymerase chain reaction (PCR) and real-time PCR targeting the putative DNA polymerase and 16S rDNA sequence of ‘Candidatus Liberibacter asiaticus,’ respectively, were used to examine the distribution and movement of the HLB pathogen in the infected citrus tree. We found that ‘Ca. Liberibacter asiaticus’ was distributed in bark tissue, leaf midrib, roots, and different floral and fruit parts, but not in endosperm and embryo, of infected citrus trees. Quantification analysis of the HLB bacterium indicated that it was distributed unevenly in planta and ranged from 14 to 137,031 cells/μg of total DNA in different tissues. A relatively high concentration of ‘Ca. Liberibacter asiaticus’ was observed in fruit peduncles. Our data from greenhouse-infected plants also indicated that ‘Ca. Liberibacter asiaticus’ was transmitted systemically from infection site to different parts of the plant. Understanding the distribution and movement of the HLB bacterium inside an individual citrus tree is critical for discerning its virulence mechanism and to develop management strategies for HLB.
Disease–Weather Relationships for Powdery Mildew and Yellow Rust on Winter Wheat
Authors: Te Beest D. E. , Paveley N. D. , Shaw M. W. , van den Bosch F.
Published: April 2008

Key weather factors determining the occurrence and severity of powdery mildew and yellow rust epidemics on winter wheat were identified. Empirical models were formulated to qualitatively predict a damaging epidemic (>5% severity) and quantitatively predict the disease severity given a damaging epidemic occurred. The disease data used was from field experiments at 12 locations in the UK covering the period from 1994 to 2002 with matching data from weather stations within a 5 km range. Wind in December to February was the most influential factor for a damaging epidemic of powdery mildew. Disease severity was best identified by a model with temperature, humidity, and rain in April to June. For yellow rust, the temperature in February to June was the most influential factor for a damaging epidemic as well as for disease severity. The qualitative models identified favorable circumstances for damaging epidemics, but damaging epidemics did not always occur in such circumstances, probably due to other factors such as the availability of initial inoculum and cultivar resistance.
Phenotypic Expression, Stability, and Inheritance of a Recessive Resistance to Monopartite Begomoviruses Associated with Tomato Yellow Leaf Curl Disease in Tomato
Authors: García-Cano E. , Resende R. O. , Boiteux L. S. , Giordano L. B. , Fernández-Muñoz R. , Moriones E.
Published: April 2008

Tomato-infecting begomoviruses comprise a complex of monopartite and bipartite virus species that cause severe yield and quality losses worldwide. Therefore, the availability of wide spectrum resistance for begomovirus control is desirable. However, limited sources of resistance are available. In this study, three tomato inbred lines with resistance to bipartite begomoviruses of Brazil were tested for resistance to monopartite begomoviruses associated with the tomato yellow leaf curl disease (TYLCD). Stable resistance to Tomato yellow leaf curl virus was observed either by inoculation with Bemisia tabaci or with Agrobacterium tumefaciens using an infectious clone. The resistance resulted in a complete absence of TYLCD symptoms and restricted virus accumulation. Further studies performed with the line ‘468-1-1-12’ indicated that the resistance was also effective against three other virus species associated with TYLCD, indicating wide spectrum resistance of this source. Quantitative genetics analyses suggested that a major recessive locus with epistatic interactions is controlling the resistance to TYLCD in ‘468-1-1-12’, which could facilitate introgression of this trait into elite tomato lines. The resistance was stable under field conditions with high TYLCD pressure. Mild symptoms could be observed in these conditions, and recovery from disease and from virus infection suggested an active host antiviral defense mechanism. The differential reaction of ‘468-1-1-12’ against a number of TYLCD-associated viruses and artificial chimeras between them allowed to identify a region of the virus genome that presumably contains a virus determinant for breaking the resistance to infection observed in ‘468-1-1-12’.
Enhancement of factor VIIa haemostatic efficacy by formulation with PEGylated liposomes
Authors: YATUV R. , DAYAN I. , CARMEL‐GOREN L. , ROBINSON M. , AVIV I. , GOLDENBERG‐FURMANOV M. , BARU M.
Published: April 2008

Summary.  Recombinant activated factor VII (rFVIIa) is an effective treatment of the haemophilia patient with inhibitors and acquired haemophilia. However, on account of its relatively short half‐life (HL), achieving therapeutic efficacy with FVIIa requires repeated injections. The development of a long‐acting FVIIa product would therefore be beneficial. The formulation of factor VIII with PEGylated liposomes (PEGLip) was previously shown to extend the bleeding‐free period in haemophilia patients. We report here an enhancement of haemostatic efficacy by similarly formulating FVIIa with PEGLip. Surface plasmon resonance analysis indicated that FVIIa binds non‐covalently but with high affinity and specificity to PEGLip. A one‐stage clotting assay demonstrated that formulation of FVIIa with PEGLip does not affect FVIIa activity and stability. A pharmacokinetic study in rats demonstrated that PEGLip formulation of FVIIa extends circulation time and results in higher FVIIa levels several hours after injection. Thromboelastography experiments indicated that PEGLip‐FVIIa induces faster clot formation and higher clot stability than standard formulated FVIIa. These results suggest that formulation of FVIIa with PEGLip may lead to a safe and effective long‐acting FVIIa that improves the care of haemophilic patients with inhibitors and acquired haemophilia.
Nitric oxide plays a key role in the platelet‐activating factor‐induced enhancement of resistance against systemic candidiasis
Authors: Kim Han‐A , Kim So‐Hee , Ko Hyun‐Mi , Choi Jung‐Hwa , Kim Kyoung‐Jin , Oh Sin‐Hye , Cho Kyoung‐Oh , Choi Il‐Whan , Im Suhn‐Young
Published: April 2008

Platelet‐activating factor (PAF) has been demonstrated to augment resistance against Candida albicans infection. In this study, the role of nitric oxide (NO) in PAF‐induced resistance in the kidneys was investigated. Pretreatment of the C. albicans‐infected mice with PAF resulted in strong expression of messenger RNA (mRNA) and the protein synthesis of inducible nitric oxide synthase (iNOS). These PAF effects were inhibited to a significant degree by pretreatment with the nuclear factor‐κB inhibitor, pyrrolidinedithiocarbamate. Pretreatment with PAF protected the mice from death caused by C. albicans infection and reduced the growth of fungus in the kidneys. The protective activity of PAF was abrogated by pretreatment with the iNOS inhibitor, aminoguanidine, and in the iNOS−/− mice. The PAF markedly increased the infiltration of neutrophils, but not macrophages, and also enhanced the mRNA expression levels of the CXC chemokine, keratinocyte‐derived chemokine, in C. albicans‐infected kidneys. These effects of PAF were attenuated in the aminoguanidine‐treated mice and the iNOS−/− mice. These data show that NO plays an important role in PAF‐induced protection against C. albicans.
TcR‐induced regulated secretion leads to surface expression of CTLA‐4 in CD4+CD25+ T cells
Authors: Catalfamo Marta , Tai Xuguang , Karpova Tatiana , McNally James , Henkart Pierre A.
Published: April 2008

In this study we show that CD4+ T cells develop a functional regulated secretory compartment after differentiation into effector cells, as shown by their increased expression and T‐cell receptor‐induced exocytosis of lysosomal and cytotoxic effector proteins. We tested the hypothesis that activation‐induced surface cytotoxic T‐lymphocyte‐associated antigen (CTLA‐4) expression in CD4+CD25+ regulatory T cells occurs via a similar regulated secretory pathway. Fluorescence microscopy showed that internal CTLA‐4 in these cells was stored in a vesicular compartment distinct from lysosomal vesicles. Rapid activation‐induced CTLA‐4 surface expression in mouse CD4+CD25+ T cells is independent of protein synthesis and Rab‐27a. When antigen‐dependent T‐cell–antigen‐presenting cell (APC) conjugates were analysed for surface distribution of CD86 on APC, a higher concentration of CD86 molecules was observed in the synapse of APC conjugated to CD4+CD25+ cells than APC conjugated to CD4+CD25− cells. These results demonstrate that fast delivery of mediators by the regulated secretory pathway in CD4+ T cells can be used to perform other functions that are not involved in cytotoxic function but that can influence/regulate other cells.
Alterations in the thymocyte phenotype of EphB‐deficient mice largely affect the double negative cell compartment
Authors: Alfaro David , Muñoz Juan José , García‐Ceca Javier , Cejalvo Teresa , Jiménez Eva , Zapata Agustín
Published: April 2008

In the present study, we have analysed the phenotype of EphB2 and/or EphB3 deficient thymocytes confirming and extending previous studies on the role of this family of molecules in T‐cell differentiation. In all mutant thymuses statistically significant reduced cell contents were observed. This reduction of thymic cellularity correlated with increased proportions of apoptotic cells, largely both double negative (DN; CD4− CD8−) and double positive (CD4+ CD8+) cells, and decreased proportions of DN cycling cells. Adult deficient thymuses also showed increased proportions of DN cells but not significant variations in the percentages of other thymocyte subsets. In absolute terms, the thymocyte number decreased significantly in all thymocyte compartments from the DN3 (CD44− CD25+) cell stage onward, without variations in the numbers of both DN1 (CD44+ CD25−) and DN2 (CD44+ CD25+) cells. Remarkably, all these changes also occurred from the 15‐day fetal EphB2 and/or EphB3 deficient mice, suggesting that adult phenotype results from the gradual accumulations of defects appearing early in the thymus ontogeny. As a reflection of thymus condition, a reduction in the number of T lymphocytes occurred in the peripheral blood and mesenteric lymph nodes, but not in spleen, maintaining the proportions of T‐cell subsets defined by CD4/CD8 marker expression, in all cases.
Technologies for value creation: an exploration of remote diagnostics systems in the manufacturing industry
Authors: Jonsson Katrin , Westergren Ulrika H , Holmström Jonny
Published: April 2008

Abstract.  With firms increasingly relying on ubiquitous computing to implement major business initiatives, it is becoming ever more necessary to understand the technological aspects of business developments. This paper analyzes the use of remote diagnostics systems in the manufacturing industry and discusses the opportunities and challenges for the early adopters. It pays specific attention to the impact on business aspects such as the value creation process consisting of relationships, roles, and architecture and the value proposal consisting of a business offer and customer value. The study shows how ubiquitous computing allows manufacturers to become remote service providers while customers can either become co‐creators of value or passive receivers of created value. Ubiquitous computing also creates possibilities for the manufacturing industry to design new kinds of business offers based on remote presence. Studying remote diagnostics systems shows that ubiquitous computing creates value when deployed in products, and not just in relation to individuals. Moreover, the design of the value‐creation process should not be limited to the single supplier or customer organization, as ubiquitous computing applications take no notice of organizational boundaries.
20‐year trends in clinical characteristics, therapy and short‐term prognosis in acute myocardial infarction according to presenting electrocardiogram: the MONICA / KORA AMI Registry (1985–2004)
Authors: Kuch B. , Heier M. , Von Scheidt W. , Kling B. , Hoermann A. , Meisinger C.
Published: April 2008

Objectives.  To examine the extent to which evidence‐based beneficial therapy is applied in practice, whether this is changing over time and is associated with improved outcomes.

Background.  Randomized trials have proved efficacy of several treatments for acute myocardial infarction (AMI) with ST‐elevation (STEMI), non‐ST‐elevation (NSTEMI) and bundle branch block (BBB).

Design and Setting.  We prospectively examined all 6748 consecutive patients with AMI aged 25–74 years hospitalized in the study region’s major clinic stratified into four time‐periods: 1985–1989 (n = 1622), 1990–1994 (n = 1588), 1995–1999 (n = 1450) and 2000–2004 (n = 2088).

Results.  The increase in numbers of AMI in the last period was mainly, but not exclusively driven by NSTEMI cases. Evidence‐based pharmacological therapy increased steeply over time. Invasive procedures increased mainly in the last period with percutaneous coronary intervention and coronary artery bypass graft performed in 30% and 15% in 1998 and 66.0% and 22%, respectively, in 2004. In‐hospital complications and 28‐day‐case fatality decreased significantly from period 1 to period 4 in all patients with AMI. Marked reductions in 28‐day‐case fatality were mostly seen in BBB patients during the last period (25.3% vs. 10.3%, P < 0.001). Of interest, the odds in 28‐day‐case fatality reduction was diminished after correction for recanalization therapy (from 0.35, 95% CI: 0.16–0.74 to 0.52, 95% CI: 0.19–1.45).

Conclusions.  Over the past 20 years, there were substantial changes in pharmacological and interventional therapies in AMI accompanied by reductions in in‐hospital complications and 28‐day‐case fatality in all infarction types with marked reductions in 28‐day‐case fatality in BBB patients. The latter observation may mainly be because of the increased use of interventional therapy.
Habitat fragmentation and genetic diversity of an endangered, migratory songbird, the golden‐cheeked warbler (Dendroica chrysoparia)
Authors: LINDSAY DENISE L. , BARR KELLY R. , LANCE RICHARD F. , TWEDDALE SCOTT A. , HAYDEN TIMOTHY J. , LEBERG PAUL L.
Published: April 2008

Landscape genetic approaches offer the promise of increasing our understanding of the influence of habitat features on genetic structure. We assessed the genetic diversity of the endangered golden‐cheeked warbler (Dendroica chrysoparia) across their breeding range in central Texas and evaluated the role of habitat loss and fragmentation in shaping the population structure of the species. We determined genotypes across nine microsatellite loci of 109 individuals from seven sites representing the major breeding concentrations of the species. No evidence of a recent population bottleneck was found. Differences in allele frequencies were highly significant among sites. The sampled sites do not appear to represent isolated lineages requiring protection as separate management units, although the amount of current gene flow is insufficient to prevent genetic differentiation. Measures of genetic differentiation were negatively associated with habitat connectivity and the percentage of forest cover between sites, and positively associated with geographic distance and the percentage of agricultural land between sites. The northernmost site was the most genetically differentiated and was isolated from other sites by agricultural lands. Fragmentation of breeding habitat may represent barriers to dispersal of birds which would pose no barrier to movement during other activities such as migration.
Mitochondrial DNA under siege in avian phylogeography
Authors: ZINK ROBERT M. , BARROWCLOUGH GEORGE F.
Published: April 2008

Mitochondrial DNA (mtDNA) has been the workhorse of research in phylogeography for almost two decades. However, concerns with basing evolutionary interpretations on mtDNA results alone have been voiced since the inception of such studies. Recently, some authors have suggested that the potential problems with mtDNA are so great that inferences about population structure and species limits are unwarranted unless corroborated by other evidence, usually in the form of nuclear gene data. Here we review the relative merits of mitochondrial and nuclear phylogeographical studies, using birds as an exemplar class of organisms. A review of population demographic and genetic theory indicates that mitochondrial and nuclear phylogeographical results ought to concur for both geographically unstructured populations and for populations that have long histories of isolation. However, a relatively common occurrence will be shallow, but geographically structured mtDNA trees—without nuclear gene corroboration—for populations with relatively shorter periods of isolation. This is expected because of the longer coalescence times of nuclear genes (approximately four times that of mtDNA); such cases do not contradict the mtDNA inference of recent isolation and evolutionary divergence. Rather, the nuclear markers are more lagging indicators of changes in population structure. A review of the recent literature on birds reveals the existence of relatively few cases in which nuclear markers contradict mitochondrial markers in a fashion not consistent with coalescent theory. Preliminary information from nuclear genes suggests that mtDNA patterns will prove to be robust indicators of patterns of population history and species limits. At equilibrium, mitochondrial loci are generally a more sensitive indicator of population structure than are nuclear loci, and mitochondrial estimates of FST‐like statistics are generally expected to exceed nuclear ones. Hence, invoking behavioural or ecological explanations of such differences is not parsimonious. Nuclear genes will prove important for quantitative estimates of the depths of haplotype trees, rates of population growth and values of gene flow.
Mountain coniferous forests, refugia and butterflies
Authors: VARGA ZOLTÁN
Published: April 2008

The boreal coniferous forests form the most extended vegetation zone of the Northern Hemisphere. As opposed to North America, they are disconnected from the mountain coniferous forests in Europe, because of the dominant east–west direction of the mountain chains. Consequently, the mountain forests show some unique characteristic features of glacial survival and postglacial history, as well. The mountain coniferous forests have numerous common floral and faunal elements with the boreal zone. However, the few unique faunal elements of the European mountain coniferous forests can be used to unravel the peculiar patterns and processes of this biome. In this issue of Molecular Ecology, Thomas Schmitt and Karola Haubrich (2008) use the relatively common and taxonomically well‐studied butterfly, the large ringlet (Erebia euryale) to identify the last glacial refugia and postglacial expansion routes.
Antifungal Immunotherapy and Immunomodulation: A Double‐hitter Approach to Deal with Invasive Fungal Infections
Authors: Hamad M.
Published: April 2008

In recent years, the incidence of life‐threatening fungal infections has dramatically increased. Despite significant developments in antifungal chemotherapy, its efficacy remains limited by the inability to sterilize infected organs and the tendency to induce resistance and cause side effects. In response to these challenges, it is now recognized that several aspects of antifungal immunity can be modulated to better deal with fungal infections. Extensive work was carried out on the development and testing of preventive and therapeutic antifungal vaccines. The potential use of cytokines, adoptive T‐cell transfer, monoclonal antibodies (MoAb) and antimicrobial peptides (AMP) as solo or adjunctive therapies is also receiving much attention. Although each of these immune‐based treatment strategies has many advantages and some shortcomings, none on its own, has proven satisfactorily effective to deal with invasive fungal infections. Appropriate combinations that optimize the advantages and minimize the disadvantages of immune‐based antifungals are still lacking mainly due to the immense difficulty in sorting out candidate combinations given the long list of choices. In this review, immune‐based antifungals are divided into two general categories on the basis of the intended target being the host (immunomodulation through vaccines, cytokines, adoptive T‐cell transfer or MoAb) or the pathogen (immunotherapy through MoAb or AMP). Potential advantages and disadvantages of immunotherapy and immunomodulation are tentatively discussed so as to facilitate the design of future studies that aim at devising more potent immune‐based antifungal treatment combinations.
The Tuberculin Skin Test Increases the Responses Measured by T Cell Interferon‐Gamma Release Assays
Authors: Vilaplana C. , Ruiz‐Manzano J. , Gil O. , Cuchillo F. , Montané E. , Singh M. , Spallek R. , Ausina V. , Cardona P. J.
Published: April 2008

RUTI is a vaccine consisting of Mycobacterium tuberculosis bacilli grown in stress conditions that is fragmented, detoxified and liposomed. RUTI was designed to shorten the treatment of latent tuberculosis infection (LTBI) with isoniazid from 9 months to just 1 month, by additional treatment with two inoculations of RUTI 4 weeks apart. During the validation process for monitoring the immunogenicity of administration of RUTI in a Phase I clinical trial, the question arose whether to introduce the tuberculin skin test (TST) in the screening of non‐LTBI volunteers. This study was designed to evaluate the effect of TST on subsequent different T‐cell interferon‐gamma release assay (TIGRA) responses, using a spectrum of M. tuberculosis‐related antigens (ESAT‐6, CFP‐10, 16 kDa, 19 kDa, MPT64, Ag 85B, 38 kDa, hsp65, PPD and BCG). The results showed an increase in post‐TST response even in non‐LTBI subjects for most antigens tested, as measured both by whole blood assay (WBA) and ELISPOT. Increased ELISPOT response decreased toward pre‐TST levels within 1 month whereas the WBA response did not. Taking into account that there is no definitive correlation between TST and TIGRA tests to diagnose LTBI and the feasibility that TST might alter the immune monitoring included in clinical trials, these data suggest that TST determination should be carefully planned to avoid any interference with TIGRA.
TLR7/8 Ligand, R‐848, Inhibits IgE Synthesis by Acting Directly on B Lymphocytes
Authors: Shen E. , Lu L. , Wu C.
Published: April 2008

TLRs are involved in the regulation of immune responses. R‐848, a TLR7/8 ligand, has potent anti‐viral and anti‐tumour properties and has been used as a new immune response modifier for enhancing Th1 immune response. In this study, we found that R‐848 significantly inhibited IgE synthesis from murine B cells at the single cell levels by anti‐CD40 plus IL‐4‐stimulated splenocytes, in which R‐848 acted on the early stage of B cell differentiation to modulate IgE synthesis. This inhibitory effect of R‐848 on IgE synthesis was not isotype specific as it also inhibited IgG1 synthesis. Moreover, R‐848 had no significant effect on the production of IgG2a by anti‐CD40 plus IL‐4‐stimulated splenocytes. Further studies showed that R‐848 markedly promoted murine B cell activation induced by anti‐CD40 plus IL‐4 by up‐regulating the expression of B cell activation markers CD25, CD69 and co‐stimulatory molecule CD80. In contrast, R‐848 inhibited the proliferation and division of murine B cells in anti‐CD40 plus IL‐4‐stimulated splenocytes. R‐848 promoted the production of IFN‐γ and IL‐12 that were partially responsible for its inhibitory effect on IgE production by anti‐CD40 plus IL‐4 because the addition of anti‐IFN‐γ or anti‐IL‐12 mAbs to the cultures could significantly restore IgE production by splenocytes. Importantly, R‐848 had a direct effect on purified B cells to inhibit IgE production induced by anti‐CD40 plus IL‐4. Taken together, these results demonstrate that R‐848 markedly inhibits IgE synthesis, and suggest that R‐848 could be used to treat allergic diseases.
Gender Variation in Interleukin‐13 Production: A Possible Mechanism of Differential In Vivo Growth of a T‐cell Lymphoma
Authors: Deepak P. , Kumar S. , Acharya A.
Published: April 2008

Interleukin (IL)‐13 is a TH2 type of cytokine that plays a crucial role in the pathophysiology of different type of infections, autoimmune diseases and malignancies. It has been shown to dampen the TH1 type of immune responses and favours tumour growth. In the present investigation, we have determined IL‐13 level in serum and ascitic fluid in both the sexes of BALB/c strain of mice bearing a T‐cell lymphoma of spontaneous origin, designated as Dalton’s Lymphoma (DL). Further, we have studied the involvement of gender hormones on the IL‐13 level and NKT‐cell production of IL‐13. It has been observed that there exists a gender variation or gender dimorphism in the IL‐13 production. IL‐13 level in serum is directly correlated with in vivo progressive growth of DL cells. We observed a tendency in female DL‐bearing mice to have higher serum IL‐13 level and faster growth of DL cells. This study, therefore, indicates that sex hormones are directly involved in the differential production of IL‐13 that may be a factor responsible, at least in part, for the differential in vivo progressive growth of a T‐cell lymphoma.
Defective in Tapetal Development and Function 1 is essential for anther development and tapetal function for microspore maturation in Arabidopsis
Authors: Zhu Jun , Chen Hui , Li Hui , Gao Ju‐Fang , Jiang Hua , Wang Chen , Guan Yue‐Feng , Yang Zhong‐Nan
Published: April 2008

In Arabidopsis, the tapetum plays important roles in anther development by providing enzymes for callose dissolution and materials for pollen‐wall formation, and by supplying nutrients for pollen development. Here, we report the identification and characterization of a male‐sterile mutant, defective in tapetal development and function 1 (tdf1), that exhibits irregular division and dysfunction of the tapetum. The TDF1 gene was characterized using a map‐based cloning strategy, and was confirmed by genetic complementation. It encodes a putative R2R3 MYB transcription factor, and is highly expressed in the tapetum, meiocytes and microspores during anther development. Callose staining and gene expression analysis suggested that TDF1 may be a key component in controlling callose dissolution. Semi‐quantitative and quantitative RT‐PCR analysis showed that TDF1 acts downstream of DYT1 and upstream of AMS and AtMYB103 in the transcriptional regulatory networks that regulate tapetal development. In conclusion, our results show that TDF1 plays a vital role in tapetal differentiation and function.