bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of a Warning System for Controlling Primary Infections of Grapevine Downy Mildew
Authors: Caffi T. , Rossi V. , Bugiani R.
Published: May 2010

A warning system based on (i) a model that simulates the development of all cohorts of Plasmopara viticola oospores, from oospore germination to infection; (ii) short-term weather forecasts; and (iii) a mobile phone short message system was tested in Northern Italy, from 2006 to 2008. An unsprayed control was compared with a “Warning A” treatment (WA, fungicides were applied whenever the warning system predicted an infection period), a “Warning B” treatment (WB, fungicides were applied as in the WA treatment but only when the relative dimension of any oospore cohort predicted by the model exceeded a threshold), and a “grower” treatment (fungicides were applied according to a conventional schedule). Average disease incidence on leaves was reduced by up to 90% in sprayed plots compared with unsprayed plots. On bunches, efficacy was always >90% at fruit set; when most berries were touching, efficacy was higher for the WA (96%) than for grower (89%) and WB (85%) treatments. On average, 6.8 fungicide sprays were applied following the grower's schedule; use of the warning system reduced applications by about one-half (WA treatment) or two-thirds (WB treatment). The grower's schedule had an average cost of 337 €/ha; the average saving with the WA and the WB treatments was 174 and 224 €/ha, respectively.
BotryosphaeriaceaeSpecies Spore-Trapping Studies in California Vineyards
Authors: Úrbez-Torres J. R. , Battany M. , Bettiga L. J. , Gispert C. , McGourty G. , Roncoroni J. , Smith R. J. , Verdegaal P. , Gubler W. D.
Published: May 2010

The seasonal abundance of Botryosphaeriaceae spp. spores was studied in California vineyards by using glass microscope slides covered with petroleum jelly placed on grapevine cordons and Burkard volumetric spore traps at seven and two different locations, respectively. Correlation analysis was used to determine which meteorological variables (precipitation, relative humidity, temperature, and wind speed) influenced Botryosphaeriaceae spp. spore release. Among all variables, regression analysis resulted in a strong relationship between spore release and precipitation. Additionally, a positive relationship between irrigation and spore release was also observed in the Riverside County vineyard. During the study period, spore discharge of Botryosphaeriaceae spp. occurred from the first fall rain through the last spring rains, coinciding with September to April. However, based on the results obtained from the spore traps, most spores (over 60%) were trapped following rain events during the winter months of December, January, and February, which coincides with the grapevine pruning season. Botryosphaeriaceae spp. spore release was much lower in fall and early spring (22%) and very few or no spores were trapped in late spring and summer (3%). This work suggests that a delay of pruning time in California may be warranted to reduce grapevine infection because the current timing coincides with the greatest period of spore discharge.
Wind Speed Effects on the Quantity of Xanthomonas citri subsp. citri Dispersed Downwind from Canopies of Grapefruit Trees Infected with Citrus Canker
Authors: Bock C. H. , Graham J. H. , Gottwald T. R. , Cook A. Z. , Parker P. E.
Published: May 2010

The epidemic of citrus canker (Xanthomonas citri subsp. citri) in Florida continues to expand since termination of the eradication program in 2006. Storms are known to be associated with disease spread, but little information exists on the interaction of fundamental physical and biological processes involved in dispersal of this bacterium. To investigate the role of wind speed in dispersal, wind/rain events were simulated using a fan to generate wind up to 19 m·s-1 and spray nozzles to simulate rain. Funnels at ground level and panels at 1.3 m height and distances up to 5 m downwind collected wind-driven splash. Greater wind speeds consistently dispersed more bacteria, measured by concentration (colony forming units [CFU] ml-1) or number sampled (bacteria flux density [BFD] = bacteria cm-2 min-1), from the canopy in the splash. The CFU ml-1 of X. citri subsp. citri collected by panels 1 m downwind at the highest wind speed was up to 41-fold greater than that collected at the lowest wind speed. BFD at the highest wind speed was up to 884-fold higher than that collected at the lowest wind speed. Both panels at distances >1 m and funnels at distances >0 m collected many-fold more X. citri subsp. citri at higher wind speeds compared to no wind (up to 1.4 × 103-fold greater CFU ml-1 and 1.8 × 105-fold the BFD). The resulting relationship between wind speed up to 19 m·s-1 and the mean CFU ml-1 collected by panel collectors downwind was linear and highly significant. Likewise, the mean CFU ml-1 collected from the funnel collectors had a linear relationship with wind speed. The relationship between wind speed and BFD collected by panels was generally similar to that described for CFU ml-1 of X. citri subsp. citri collected. However, BFD collected by funnels was too inconsistent to determine a meaningful relationship with increasing wind speed. The quantity of bacteria collected by panels declined with distance, and the relationship was described by an inverse power model (R2 = 0.94 to 1.00). At higher wind speeds, more bacteria were dispersed to all distances. Windborne inoculum in splash in subtropical wet environments is likely to be epidemiologically significant, as both rain intensity and high wind speed can interact to provide conditions conducive for dispersing large quantities of bacteria from canker-infected citrus trees. Disease and crop management aimed at reducing sources of inoculum and wind speeds in a grove should help minimize disease spread by windborne inoculum.
Effects of Wheat streak mosaic virus on Root Development and Water-Use Efficiency of Hard Red Winter Wheat
Authors: Price J. A. , Workneh F. , Evett S. R. , Jones D. C. , Arthur J. , Rush C. M.
Published: May 2010

Greenhouse and field studies were conducted to determine the effects of Wheat streak mosaic virus (WSMV), a member of the family Potyviridae, on root development and water-use efficiency (WUE) of two hard red winter wheat (Triticum aestivum) cultivars, one susceptible and one resistant to WSMV. In the greenhouse studies, wheat cultivars were grown under three water regimes of 30, 60, and 80% soil saturation capacity. After inoculation with WSMV, plants were grown for approximately 4 weeks and then harvested. Root and shoot weights were measured to determine the effect of the disease on biomass. In all water treatments, root biomass and WUE of inoculated susceptible plants were significantly less (P < 0.05) than those of the noninoculated control plants. However, in the resistant cultivar, significance was only found in the 30 and 60% treatments for root weight and WUE, respectively. Field studies were also conducted under three water regimes based on reference evapotranspiration rates. Significant reductions in forage, grain yield, and crop WUE were observed in the inoculated susceptible plots compared with the noninoculated plots. Both studies demonstrated that wheat streak mosaic reduces WUE, which is a major concern in the Texas Panhandle because of limited availability of water.
Physiologic Specialization of Puccinia triticina on Wheat in the United States in 2008
Authors: Kolmer J. A. , Long D. L. , Hughes M. E.
Published: May 2010

Collections of Puccinia triticina were obtained from rust-infected wheat (Triticum aestivum) leaves provided by cooperators throughout the United States and from surveys of wheat fields and wheat breeding plots by USDA-ARS personnel in the Great Plains, Ohio River Valley, Southeast, and Washington State in order to determine the virulence of the wheat leaf rust population in 2008. Single uredinial isolates (730 in total) were derived from the collections and tested for virulence phenotype on lines of Thatcher wheat that are near-isogenic for leaf rust resistance genes Lr1, Lr2a, Lr2c, Lr3, Lr9, Lr16, Lr24, Lr26, Lr3ka, Lr11, Lr17, Lr30, LrB, Lr10, Lr14a, Lr18, Lr21, Lr28, and a winter wheat line with Lr41. Forty-eight virulence phenotypes were described. Virulence phenotypes TDBGG, TCRKG, and MLDSD were the three most common phenotypes. TDBGG is virulent to Lr24 and was found in both the soft red winter wheat and hard red winter wheat regions. Phenotype TCRKG is virulent to Lr11, Lr18, and Lr26 and is found mostly in the soft red winter wheat region in the eastern United States. Phenotype MLDSD is virulent to Lr17 and Lr41 and was widely distributed in the Great Plains. Virulence to Lr21 was not found in any of the tested isolates. Virulence to Lr11 and Lr18 increased in 2008 in the soft red winter wheat regions. Two separate epidemiological zones of P. triticina in the soft red winter wheat region of the southern and eastern states and in the hard red wheat region of the Great Plains were described.
First Report of Armillaria Root Disease Caused by Armillaria tabescens on Araucaria araucana in Veracruz, Mexico
Authors: Kim M.-S. , Klopfenstein N. B. , Hanna J. W. , Cannon P. , Medel R. , López A.
Published: May 2010

In September 2007, bark samples were collected from the root collar of a single Araucaria araucana tree that had recently died and was suspected of being killed by Armillaria root disease. Disease symptoms and signs included a thinning crown and fruiting bodies at the tree base over a several-year period before tree death. The tree was located in an isolated street-tree planting within a business district on Maestros Veracruzanos Street, Xalapa, Veracruz (19°31′52′′N, 96°54′25′′W, elevation 1,392 m). One fungal isolate (MEX21WF) was obtained, which possessed two sequence repeat types from the intergenic spacer-1 (IGS-1) region (GenBank Accession Nos. GQ335541 and GQ335542). On the basis of these IGS-1 sequences, this isolate from Mexico possessed 99% nucleotide sequence identities with North American Armillaria tabescens isolates (GenBank Accession Nos. AY695410 ≈ GQ335541 and AY773966 ≈ GQ335542). Somatic pairing tests of the isolate with other North American Armillaria species also identified it as A. tabescens (2). In addition, fruiting bodies were produced on the stump base in 2009 that matched morphological features of A. tabescens, e.g., exannulate, cespitose growth in clusters, brown-gray stipe to blackish toward the base, longitudinally fibrillose, basidiospores (6–) 7 to 9 × 4 to 5 (–5.5) μm, and other general morphology. On the basis of these three lines of taxonomic evidence, it was concluded that the isolate was A. tabescens. To our knowledge, this is the first confirmed report of A. tabescens causing Armillaria root disease in Mexico. Furthermore, this note represents the first report of A. tabescens on Araucaria araucana, which is native to Chile and Argentina. The other previous reports of A. tabescens in Mexico are based on herbarium specimens collected in 1965 (BPI 753040) from Valle de Bravo (approximately 350 km west of Xalapa) in the state of México and 1973 (BPI 753041) from near Monterrey (approximately 760 km north-northwest of Xalapa) in the state of Nuevo León (1). However, no host information or confirmation of taxonomic identification was reported for these herbarium specimens. Although this note confirms the presence of A. tabescens in Mexico, more surveys and monitoring are needed to determine the full distribution of this pathogen in Mexico. Because the climate and tree communities of eastern Mexico are similar to those of the southeastern United States, where A. tabescens has been reported as a common pathogen of oaks and fruit trees (3,4), it seems reasonable that A. tabescens may represent an existing or potential threat in eastern Mexico.

References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory. Online publication. ARS, USDA, 2010. (2) K. I. Mallett and Y. Hiratsuka. Can. J. Bot. 64:2588, 1986. (3) F. Miranda and A. J. Sharp. Ecology 31:313, 1950. (4) G. Schnabel et al. Mycol. Res. 109:1208, 2005.
First Report of Iris yellow spot virus on Onion in Uruguay
Authors: Colnago P. , Achigar R. , González P. H. , Peluffo S. , González Idiarte H. , Pianzzola M. J. , Galván G. A.
Published: May 2010

From October to December 2005, onion (Allium cepa) plants in seed-production fields in south Uruguay (Canelones) had symptoms suggestive of those caused by Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae). Symptoms included diamond-shaped lesions on seed stalks (scapes), each 1 to 5 cm long with a necrotic border, green center, and sometimes a second necrotic area in the center of the diamond (2,3). Necrotic lesions with more irregular shape were also associated with diseased plants. In 2006, scape samples with these symptoms were collected from four onion seed crops and assayed for IYSV using an IYSV-specific antiserum (Agdia Inc., Elkhart, IN) in a double-antibody sandwich-ELISA. IYSV was detected in all four onion seed crops monitored in 2006. IYSV incidence, expressed as the number of plants with symptoms, ranged from <1% (1 of 120 plants evaluated) to 20% (24 of 120 plants). Two fields were monitored in 2007, in which IYSV incidence increased from 2 and 3% in October to 7% (198 of 2,768 plants) and 40% (253 of 638 plants) in December, respectively. The highest incidence was observed in the same farm in 2006 and 2007. Scapes were sampled from the field with the highest incidence of symptoms in 2007 and tested for IYSV with IYSV-specific primers (3). Total RNA was extracted from 100 mg of symptomatic tissue, with green tissue adjacent to typical lesions, following a Trizol-based protocol (1). A reverse transcriptase-PCR assay with nucleocapsid gene-specific primers was used (3). A PCR product of approximately 26 bp was obtained, coincident with the expected length for IYSV. The PCR product was cloned and sequenced. The tospovirus N sequence of the isolate in Uruguay (Accession No. GU550518) had maximum identity (97%) with an Australian IYSV isolate (Accession No. AY345227), and >87% identity only with IYSV N protein sequences in GenBank. Because of the presence of IYSV in Brazil, Chile, and Peru, this first documentation, to our knowledge, of IYSV in onion crops in Uruguay suggests that the threat of IYSV to onion is increasing in South America.

References: (1) P. Chomczynski and K. Mackey. Biotechniques 19:942, 1995. (2) D. H. Gent et al. Plant Dis. 90:1468, 2006. (3) H. R. Pappu et al. Plant Dis. 92:588, 2008.
Pequi is a Host for Pratylenchus zeae in the Brazilian Cerrado Region
Authors: Machado A. C. Z. , Bonfim M. F. , de Araújo Filho J. V.
Published: May 2010

The Brazilian Cerrado Region has many natural resources that have high social economic interest. Pequi (Caryocar brasiliense Camb.), a native species from that area, has an edible fruit, which is highly appreciated by the local population, and also a high-quality wood. In January 2010, pequi root samples were collected near the municipality of Rio Verde, Goiás State, Brazil (17°49′25.76″S, 51°02′10.06″W). Roots were washed with tapwater, dried on absorbent paper, cut in 1-cm2 pieces, and processed for nematode extraction by the blender centrifugal flotation method (2). The specimens were identified by morphological and morphometrical characteristics of six adult females mounted in formaldehyde temporary slides (1). Morphological characters used for identification included female body, stylet, pharyngeal overlapping, pharynges, postvulval uterine sac, tail lengths, stylet knobs, number of labial rings, vulva position in relation to body length, body diameters (high body, vulval, and anus region), and the de Man's ratios (a, b, b', c, and c'). Characters measured were consistent with those described for Pratylenchus zeae Graham, 1951 (1); the labial region showed three annuli, stylet was 14.83 (±0.93) μm long, with broad, anteriorly flattened basal knobs. Vulva position was 71.66% (±0.98) of body length and spermatheca was round, small, and without sperm (males were not found). Postvulval uterine sac was short (31.3 ± 4.03 μm) and tail (26.6 ± 3.61 μm) was conoid, pointed, and unstriated. Pharyngeal overlapping length was 30.5 (±6.5) μm; pharynges were 150.83 (± 28.16) μm long. The de Man's ratios obtained were: a = 24.26 ± 2.31; b = 3.89 ± 0.69; b' = 3.08 ± 0.48; c = 17.17 ± 1.47; and c' = 2.25 ± 0.19. To our knowledge, this is the first report of P. zeae infecting pequi. It is difficult to determine the economic importance of this nematode parasite to pequi production since pequi is not yet a commercial crop in Brazil. This finding, however, has long term importance because researchers have been developing improved cultivars by combining favorable agronomic characteristics with high oilseed content for biofuel production. If these are commercialized, P. zeae could become an important pathogen in pequi plantings.

References: (1) P. Castillo and N. Vovlas. Pratylenchus (Nematoda: Pratylenchidae): Diagnosis, Biology, Pathogenicity and Management. Koninklijke Brill NV, Leiden, the Netherlands, 2007. (2) W. A. Coolen and C. J. D'Herde. A Method for the Quantiative Extraction of Nematodes from Plant Tissue. State Agric. Entomol. Res. Stn. Ghent, Belgium, 1972.
Occurrence of Poinsettia Stem Flat Disease Caused by Phytoplasma in Korea
Authors: Chung B. N. , Choi G. S.
Published: May 2010

In December 2009, commercially grown poinsettia (Euphorbia pulcherrima Willd cv. Ichibang) exhibited typical phytoplasma-like symptoms in 95% of an affected field in Yongin, Korea (Gyeonggi Province). Symptoms consisted of flat stems and fascicles and an abnormal number of apexes resulting in a cockscomb form of stem and flower bud proliferation. Leaf narrowing with curling of bracts was also associated with the disease. Symptomatic poinsettia plants were not marketable. Poinsettias, cv. Ichibang, without symptoms were obtained from a breeding collection in a glasshouse of the National Institute of Horticultural and Herbal Science in Suwon (Gyeonggi Province). The presence of phytoplasmas in symptomatic and healthy-looking flowers and stems of cv. Ichibang was demonstrated by PCR analysis with primer pair R16mF2/R16mR1 (1), which amplifies phytoplasma 16S rDNA regions. PCR products (~1,427 bp), were obtained from both symptomatic and healthy-looking plants, sequenced, and registered under the GenBank Accession Nos. of GU461275 and GU461277, respectively. Symptomatic poinsettia and healthy poinsettia sequences had 99.6 and 100% identity with U.S. PoiBI isolate FJ376625, indicating poinsettia stem flat disease is caused by PoiBI. A branch-inducing factor in poinsettia has been known for several decades, but only since 1997 was this graft transmissible factor identified as a PoiBI phytoplasma (1), belonging to the peach X-disease phytoplasma group (16S rRNA III) subgroup 16SrIII-H (2). Normally, phytoplasmas are associated with host quality loss, but abnormally, infection in poinsettia generates a desirable, free-branching growth habit. In this study we found that PoiBI could be detrimental to the quality of poinsettia depending on the cultivar and agronomic practice. Poinsettia stem flat disease presumably occurred because of increased levels of phytoplasma caused by successive stem cutting for commercial use. To our knowledge, this is the first report of PoiBI phytoplasma that affected marketability of poinsettia in Korea.

References: (1) I. M. Lee et al. Nature Biotech. 15:178, 1997. (2) I. M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.
First Report of Sclerotinia Stem Rot of Canola Caused by Sclerotinia sclerotiorum in Texas
Authors: Isakeit T. , Woodward J. E. , Niu C. , Wright R. J.
Published: May 2010

During the past several years, canola (Brassica napus L.) has been grown experimentally in different areas of Texas to evaluate its potential as a crop, particularly for use as a biofuel source. In early April 2007, symptoms typical of Sclerotinia stem rot were observed in a canola variety trial that was flowering in Wharton County, Texas. Stems had white mycelia growing on the outside, or a bleached appearance, near the soil surface and plants were lodging. Inside bleached stems, there were spherical to cylindrical, black sclerotia that were 3 to 10 mm. Isolations from surface-disinfested stems onto potato dextrose agar (PDA) consistently yielded white, fluffy colonies with sclerotia typical of Sclerotinia sclerotiorum (Lib.) de Bary (1). Sequence analyses were conducted on two replicates of mycelium by extracting fungal DNA with the Qiagen DNeasy Plant Mini Kit (Valencia, CA). PCR amplification was performed using two primer sequences (92-4 AF377919: TCGCCTCAGAAGAATGTGC/AGCGGGTTACAAGGAGATGG; and 119-4 AF377925: GTAACAAGAGACCAAAATTCGG/TGAACGAGCTGTCATTCCC) (2) that have previously been used to characterize S. sclerotiorum (3). The BLAST search revealed that the sequences were 99 and 98% homologous with S. sclerotiorum Accession Nos. AF377919 and AF377925 over 376 and 377 bp of aligned sequence, respectively. Agar segments (1 cm2) from a 5-day-old culture grown on PDA were placed in the leaf axils of 15 2-month-old canola plants (‘Wichita’) growing in pots. Plants were placed in a humid chamber under fluorescent lights at 16 to 22°C. After 2 days, water soaking and necrosis occurred on petioles and stems adjacent to the inoculum, but not on plants treated with sterile PDA. S. sclerotiorum was consistently reisolated from symptomatic tissue plated on acidified PDA. The inoculations were repeated once with similar results. To our knowledge, this is the first report of Sclerotinia stem rot of canola in Texas. Currently, there is no significant canola production in Texas; however, interest in biofuels could lead to an increase in planted acres. Sclerotinia stem rot of canola could become a significant disease problem in areas of Texas where canola is planted as a winter crop.

References: (1) L. M. Kohn. Phytopathology 69:881, 1979. (2) C. Sirjusingh and L. M. Kohn. Mol. Ecol. Notes 1:267, 2001. (3) J. E. Woodward et al. Plant Dis. 92:1468, 2008.
Characterization of Plant Carotenoid Cyclases as Members of the Flavoprotein Family Functioning with No Net Redox Change
Authors: Mialoundama Alexis Samba , Heintz Dimitri , Jadid Nurul , Nkeng Paul , Rahier Alain , Deli Jozsef , Camara Bilal , Bouvier Florence
Published: May 2010

The later steps of carotenoid biosynthesis involve the formation of cyclic carotenoids. The reaction is catalyzed by lycopene β-cyclase (LCY-B), which converts lycopene into β-carotene, and by capsanthin-capsorubin synthase (CCS), which is mainly dedicated to the synthesis of κ-cyclic carotenoids (capsanthin and capsorubin) but also has LCY-B activity. Although the peptide sequences of plant LCY-Bs and CCS contain a putative dinucleotide-binding motif, it is believed that these two carotenoid cyclases proceed via protic activation and stabilization of resulting carbocation intermediates. Using pepper (Capsicum annuum) CCS as a prototypic carotenoid cyclase, we show that the monomeric protein contains one noncovalently bound flavin adenine dinucleotide (FAD) that is essential for enzyme activity only in the presence of NADPH, which functions as the FAD reductant. The reaction proceeds without transfer of hydrogen from the dinucleotide cofactors to β-carotene or capsanthin. Using site-directed mutagenesis, amino acids potentially involved in the protic activation were identified. Substitutions of alanine, lysine, and arginine for glutamate-295 in the conserved 293-FLEET-297 motif of pepper CCS or LCY-B abolish the formation of β-carotene and κ-cyclic carotenoids. We also found that mutations of the equivalent glutamate-196 located in the 194-LIEDT-198 domain of structurally divergent bacterial LCY-B abolish the formation of β-carotene. The data herein reveal plant carotenoid cyclases to be novel enzymes that combine characteristics of non-metal-assisted terpene cyclases with those attributes typically found in flavoenzymes that catalyze reactions, with no net redox, such as type 2 isopentenyl diphosphate isomerase. Thus, FAD in its reduced form could be implicated in the stabilization of the carbocation intermediate.
Significance of alternatively activated macrophages in patients with intrahepatic cholangiocarcinoma
Authors: Hasita Horlad , Komohara Yoshihiro , Okabe Hirohisa , Masuda Toshiro , Ohnishi Koji , Lei Xiao F. , Beppu Toru , Baba Hideo , Takeya Motohiro
Published: May 2010

Many studies have shown that tumor‐associated macrophages (TAMs) contribute to tumor development and poor prognosis in various cancers. In this study, we investigated the macrophage populations and phenotypes, and their correlation to angiogenesis, immunosuppression, and clinical prognosis in intrahepatic cholangiocarcinoma (ICC). CD68 (+) and CD163 (+) macrophage infiltration was analyzed in paraffin‐embedded tissue samples from 39 patients. CD163 is used as a marker of M2 macrophages. Neovascularization and infiltration of forkhead box P3 (FOXP3) (+) regulatory T cells were also evaluated. The number of CD68 (+) and CD163 (+) macrophages was positively correlated with the numbers of vessels and regulatory T cells. The number of CD163 (+) cells was more closely associated with them. Intrahepatic cholangiocarcinoma (ICC) patients with high counts of CD163 (+) macrophages showed poor disease‐free survival (P = 0.0426). The macrophage density was not correlated with overall survival. In an in vitro study using ICC cell lines (HuCCT1, RBE, and MEC) and human macrophages, tumor cell supernatant (TCS) from cell lines induced an activation of signal transducers and activators of transcription‐3 (Stat3) and macrophage polarization toward the M2 phenotype. Tumor cell supernatant (TCS) from HuCCT1 most strongly induced Stat3 activation and production of cytokines and other bioactive molecules such as interleukin (IL)‐10, vascular endothelial growth factor (VEGF)‐A, transforming growth factor (TGF)‐β, and matrix metalloproteinase (MMP)‐2. Down‐regulation of Stat3 by siRNA significantly suppressed the production of IL‐10 and VEGF‐A. These results provide suggestive evidence that TAMs contribute to cancer progression via Stat3 activation, and CD163 is useful for evaluating M2 TAMs and predicting the clinical prognosis of ICC patients. (Cancer Sci)
Osteopontin is a new target molecule for ovarian clear cell carcinoma therapy
Authors: Matsuura Motoki , Suzuki Takahiro , Saito Tsuyoshi
Published: May 2010

Recent studies have demonstrated overexpression of osteopontin (OPN) in ovarian clear cell carcinoma. Here, we revealed the role of OPN in invasiveness in ovarian clear cell carcinoma. We used immunofluorescence analysis to detect OPN in a total of 160 patient‐derived specimens. Ovarian clear cell carcinoma cell lines, RMG‐1 and TOV‐21G, were used to monitor changes in OPN and integrin levels, and cell invasiveness following treatment with OPN, simvastatin, and transfection with siRNA. Immunofluorescence analysis revealed statistically significant differences among the histological groups, and ovarian clear cell carcinoma expressed a strong OPN signal. The OPN receptors, alpha v and 5, and beta 1 and 3 integrins, were increased after treatment with OPN. Invasion assays indicated that OPN enhanced in vitro extracellular matrix invasion dose‐dependently in ovarian clear cell carcinoma. Simvastatin significantly reduced expression of OPN and the integrins, and decreased ECM invasion. RNA interference also suppressed ECM invasion. These results suggest that down‐ or up‐regulation of OPN is involved in carcinoma cell invasion. We thus conclude that OPN regulation could have a crucial role in ovarian clear cell carcinoma therapy. (Cancer Sci 2010)
MicroRNA‐10b regulates tumorigenesis in neurofibromatosis type 1
Authors: Chai Guolin , Liu Ning , Ma Junrong , Li Hua , Oblinger Janet L. , Prahalad Agasanur K. , Gong Meng , Chang Long‐Sheng , Wallace Margaret , Muir David , Guha Abhijit , Phipps Roger J. , Hock Janet M. , Yu Xijie
Published: May 2010

MicroRNAs (miRNAs) are frequently deregulated in human tumors, and play important roles in tumor development and progression. The pathological roles of miRNAs in neurofibromatosis type 1 (NF1) tumorigenesis are largely unknown. We demonstrated that miR‐10b was up‐regulated in primary Schwann cells isolated from NF1 neurofibromas and in cell lines and tumor tissues from malignant peripheral nerve sheath tumors (MPNSTs). Intriguingly, a significantly high level of miR‐10b correlated with low neurofibromin expression was found in a neuroectodermal cell line: Ewing’s sarcoma SK‐ES‐1 cells. Antisense inhibiting miR‐10b in NF1 MPNST cells reduced cell proliferation, migration and invasion. Furthermore, we showed that NF1 mRNA was the target for miR‐10b. Overexpression of miR‐10b in 293T cells suppressed neurofibromin expression and activated RAS signaling. Antisense inhibition of miR‐10b restored neurofibromin expression in SK‐ES‐1 cells, and decreased RAS signaling independent of neurofibromin in NF1 MPNST cells. These results suggest that miR‐10b may play an important role in NF1 tumorigenesis through targeting neurofibromin and RAS signaling. (Cancer Sci 2010)
Tomato cultivar tolerant to Tomato leaf curl New Delhi virus infection induces virus‐specific short interfering RNA accumulation and defence‐associated host gene expression
Authors: SAHU PRANAV PANKAJ , RAI NEERAJ K. , CHAKRABORTY SUPRIYA , SINGH MAJOR , CHANDRAPPA PRASANNA H. , RAMESH BANDARUPALLI , CHATTOPADHYAY DEBASIS , PRASAD MANOJ
Published: May 2010

Tomato leaf curl New Delhi virus (ToLCNDV) infection causes significant yield loss in tomato. The availability of a conventional tolerance source against this virus is limited in tomato. To understand the molecular mechanism of virus tolerance in tomato, the abundance of viral genomic replicative intermediate molecules and virus‐directed short interfering RNAs (siRNAs) by the host plant in a naturally tolerant cultivar H‐88‐78‐1 and a susceptible cultivar Punjab Chhuhara at different time points after agroinfection was studied. We report that less abundance of viral replicative intermediate in the tolerant cultivar may have a correlation with a relatively higher accumulation of virus‐specific siRNAs. To study defence‐related host gene expression in response to ToLCNDV infection, the suppression subtractive hybridization technique was used. A library was prepared from tolerant cultivar H‐88‐78‐1 between ToLCNDV‐inoculated and Agrobacterium mock‐inoculated plants of this cultivar at 21 days post‐inoculation (dpi). A total of 106 nonredundant transcripts was identified and classified into 12 different categories according to their putative functions. By reverse Northern analysis and quantitative real‐time polymerase chain reaction (qRT‐PCR), we identified the differential expression pattern of 106 transcripts, 34 of which were up‐regulated (>2.5‐fold induction). Of these, eight transcripts showed more than four fold induction. qRT‐PCR analysis was carried out to obtain comparative expression profiling of these eight transcripts between Punjab Chhuhara and H‐88‐78‐1 on ToLCNDV infection. The expression patterns of these transcripts showed a significant increase in differential expression in the tolerant cultivar, mostly at 14 and 21 dpi, in comparison with that in the susceptible cultivar, as analysed by qRT‐PCR. The probable direct and indirect relationship of siRNA accumulation and up‐regulated transcripts with the ToLCNDV tolerance mechanism is discussed.
Clinical effectiveness of leucoreduced, pooled donor platelet concentrates, stored in plasma or additive solution with and without pathogen reduction
Authors: Kerkhoffs Jean‐Louis H. , Van Putten Wim L. J. , Novotny Viera M. J. , Te Boekhorst Peter A.W. , Schipperus Martin R. , Zwaginga Jaap Jan , Van Pampus Lizzy C. M. , De Greef Georgine E. , Luten Marleen , Huijgens Peter C. , Brand Anneke , Van Rhenen Dick J. ,   on behalf of the Dutch – Belgian HOVON cooperative group
Published: May 2010

Pathogen reduction (PR) of platelet products increases costs and available clinical studies are equivocal with respect to clinical and haemostatic effectiveness. We conducted a multicentre, open‐label, randomized, non‐inferiority trial comparing the clinical effectiveness of buffy‐coat derived leucoreduced platelet concentrates (PC) stored for up to 7 d in plasma with platelets stored in platelet additive solution III (PASIII) without and with treatment with amotosalen‐HCl/ultraviolet‐A (UVA) photochemical pathogen reduction (PR‐PASIII). Primary endpoint of the study was 1‐h corrected count increment (CCI). Secondary endpoints were 24‐h CCI, bleeding, transfusion requirement of red cells and PC, platelet transfusion interval and adverse transfusion reactions. Compared to plasma‐PC, in the intention to treat analysis of 278 evaluable patients the mean difference for the 1‐h CCI of PR‐PASIII‐PC and PASIII‐PC was −31% (P < 0·0001) and −9% (P = n.s.), respectively. Twenty‐seven patients (32%) had bleeding events in the PR‐PASIII arm, as compared to 19 (19%) in the plasma arm and 14 (15%) in the PASIII arm (P = 0·034). Despite the potential advantages of pathogen (and leucocyte) inactivation of amotosalen‐HCl/UVA‐treated platelet products, their clinical efficacy is inferior to platelets stored in plasma, warranting a critical reappraisal of employing this technique for clinical use.
Evidence for a combination of pre‐adapted traits and rapid adaptive change in the invasive plant Centaurea stoebe
Authors: Henery Martin L. , Bowman Gillianne , Mráz Patrik , Treier Urs A. , Gex‐Fabry Emilie , Schaffner Urs , Müller‐Schärer Heinz
Published: May 2010

1. Introduced plants have the potential to rapidly evolve traits of ecological importance that may add to their innate potential to become invasive. During invasions, selection may favour genotypes that are already pre‐adapted to conditions in the new habitat and, over time, alter the characteristics of subsequent generations.

2. Spotted knapweed (Centaurea stoebe) occurs in two predominantly spatially separated cytotypes in its native range (Europe–Western Asia), but currently only the tetraploid form has been confirmed in the introduced range (North America), where it is invasive. We used several common garden experiments to examine, across multiple populations, whether tetraploids and diploids from the native range differ in life cycle, leaf traits and reproductive capacity and if such differences would explain the predominance of tetraploids and their advance into new habitats in the introduced range. We also compared the same traits in tetraploids from the native and introduced range to determine whether any rapid adaptive changes had occurred since introduction that may have enhanced invasive potential of the species in North America.

3. We found tetraploids had lower specific leaf area, less lamina dissection and fewer, narrower leaves than diploids. Diploids exhibited a monocarpic life cycle and produced few if any accessory rosettes. Diploids produced significantly more seeds per capitulum and had more capitula per plant than tetraploids. In contrast, the vast majority of European tetraploids continued to flower in both seasons by regenerating from multiple secondary rosettes, demonstrating a predominantly polycarpic life cycle.

4. During early growth tetraploids from North America achieved greater biomass than both tetraploids and diploids from the native range but this did not manifest as larger above‐ground biomass at maturity. In North American tetraploids there was also evidence of a shift towards a more strictly polycarpic life cycle, less leaf dissection, greater carbon investment per leaf, and greater seed production per capitulum.

5. Synthesis. Our results suggest that the characteristics of tetraploid C. stoebe pre‐adapted them (compared to diploid conspecifics) for spread and persistence of the species into habitats in North America characterized by a more continental climate. After the species’ introduction, small but potentially important shifts in tetraploid biology have occurred that may have contributed significantly to successful invasion.
The impact of lianas on 10 years of tree growth and mortality on Barro Colorado Island, Panama
Authors: Ingwell Laura L. , Joseph Wright S. , Becklund Kristen K. , Hubbell Stephen P. , Schnitzer Stefan A.
Published: May 2010

1. Lianas compete intensely with trees, but few studies have examined long‐term effects of liana infestation on tree growth and mortality. We quantified the effects of lianas in tree crowns (n = 2907) and rooted within 2 m of trees (n = 1086) on growth and mortality of 30 tree species from 1995 to 2005 on Barro Colorado Island (BCI), Panama, documented liana infestation in tree crowns in 1996 and 2007 to determine the dynamics of liana infestation, and quantified liana infestation in the crowns of 3231 additional canopy trees (d.b.h. ≥20 cm) in 2007 to compare with the same metric determined by previous studies in 1967 and 1980.

2. Severe liana infestation increased tree mortality: 21% of liana‐free trees in 1996 had died by 2007, whereas 42% of trees with more than 75% of the crown infested by lianas in 1996 had died by 2007.

3. Liana infestation of tree crowns significantly reduced tree growth, particularly on sun‐exposed trees. The proximity of rooted lianas significantly reduced the growth of shaded trees.

4. Liana infestation was dynamic: 10.9% of trees with severe liana infestation in their crowns in 1996 had shed all of their lianas by 2007 and 5.3% of trees with no lianas in their crown in 1996 had severe liana infestation in 2007.

5. Liana infestation was common: lianas were present in 53% of trees of the 30 focal species. Including lianas rooted within 2 m of the tree increased this percentage to 78%. Using both above‐ and below‐ground measures may provide a better estimate of liana competition than either measure alone.

6. Liana infestation is increasing on BCI. Lianas were present in the crowns of 73.6% of canopy trees (d.b.h. ≥20 cm). Liana canopy infestation was 57% higher than in 1980 and 65% higher than in 1967, which is consistent with reported increases in liana abundance, biomass, and leaf and flower production.

7. Synthesis. We used one of the largest studies ever conducted on lianas to confirm the negative effects of lianas on tree growth and survival over 10 years. Liana infestation of trees was widespread, dynamic and increasing on BCI.
Gonadal and Nongonadal Regulation of Sex Differences in Hypothalamic Kiss1 Neurones
Authors: Kauffman A. S.
Published: May 2010

The brains of males and females differ anatomically and physiologically, including sex differences in neurone size or number, synapse morphology and specific patterns of gene expression. Brain sex differences may underlie critical sex differences in physiology or behaviour, including several aspects of reproduction, such as the timing of sexual maturation (earlier in females than males) and the ability to generate a preovulatory gonadotrophin surge (in females only). The reproductive axis is controlled by afferent pathways that converge upon forebrain gonadotrophin‐releasing hormone (GnRH) neurones, but GnRH neurones are not sexually dimorphic. Although most reproductive sex differences probably reflect sex differences in the upstream circuits and factors that regulate GnRH secretion, the key sexually‐dimorphic factors that influence reproductive status have remained poorly defined. The recently‐identified neuropeptide kisspeptin, encoded by the Kiss1 gene, is an important regulator of GnRH secretion, and Kiss1 neurones in rodents are sexually dimorphic in specific hypothalamic populations, including the anteroventral periventricular nucleus–periventricular nucleus continuum (AVPV/PeN) and the arcuate nucleus (ARC). In the adult AVPV/PeN, Kiss1 neurones are more abundant in females than males, representing a sex difference that is regulated by oestradiol signalling during critical periods of postnatal and pubertal development. By contrast, Kiss1 neurones in the ARC are not sexually differentiated in adult rodents but, in mice, the regulation of ARC Kiss1 cells by gonadal hormone‐independent factors is sexually dimorphic during prepubertal development. These various sex differences in hypothalamic Kiss1 neurones may relate to known sex differences in reproductive physiology, such as puberty onset and positive feedback.
Developmental expression of dnaA is required for sporulation and timing of fruiting body formation in Myxococcus xanthus
Authors: Rosario Christopher J. , Singer Mitchell
Published: May 2010

Inhibition of DNA replication within the first 6 h of development results in a block in the developmental programme in the social soil bacterium Myxococcus xanthus. We have interpreted these data to imply that M. xanthus requires a new round of DNA replication early in the developmental programme. To further understand the role of DNA replication during development in M. xanthus we focused on the regulation of dnaA which encodes the initiator protein of DNA replication. In this work, we demonstrate that immediately upon nutrient deprivation dnaA (MXAN1001) transcript levels decrease to 10–15% of vegetative levels and then transiently increase between 4 and 6 h post initiation. This expression is dependent on several early developmental regulators, including relA (MXAN3204), sigD (MXAN2957) and sdeK (MXAN1014). It is also dependent upon an 85 bp region located just upstream to the dnaA promoter. Our data suggest that while developmental dnaA expression is not essential for development, its expression allows for the proper timing and maximum efficiency of the sporulation process. In addition, we speculate that developmental control of dnaA expression may provide a mechanism for predetermination of cell fate during the differentiation process.
Functional domain analysis of the Mycoplasma pneumoniae co‐chaperone TopJ
Authors: Cloward Jason M. , Krause Duncan C.
Published: May 2010

Colonization of conducting airways of humans by the prokaryote Mycoplasma pneumoniae is mediated by a differentiated terminal organelle important in cytadherence, gliding motility and cell division. TopJ is a predicted J‐domain co‐chaperone also having domains unique to mycoplasma terminal organelle proteins and is essential for terminal organelle function, as well as stabilization of protein P24, which is required for normal initiation of terminal organelle formation. J‐domains activate the ATPase of DnaK chaperones, facilitating peptide binding and proper protein folding. We performed mutational analysis of the predicted J‐domain, central acidic and proline‐rich (APR) domain, and C‐terminal domain of TopJ and assessed the phenotypic consequences when introduced into an M. pneumoniae topJ mutant. A TopJ derivative with amino acid substitutions in the canonical J‐domain histidine–proline–aspartic acid motif restored P24 levels but not normal motility, morphology or cytadherence, consistent with a J‐domain co‐chaperone function. In contrast, TopJ derivatives having APR or C‐terminal domain deletions were less stable and failed to restore P24, but resulted in normal morphology, intermediate gliding motility and cytadherence levels exceeding that of wild‐type cells. Results from immunofluorescence microscopy suggest that both the APR and C‐terminal domains, but not the histidine–proline–aspartic acid motif, are critical for TopJ localization to the terminal organelle.
Interaction specificity, toxicity and regulation of a paralogous set of ParE/RelE‐family toxin–antitoxin systems
Authors: Fiebig Aretha , Castro Rojas Cyd Marie , Siegal‐Gaskins Dan , Crosson Sean
Published: May 2010

Toxin–antitoxin (TA) gene cassettes are widely distributed across bacteria, archaea and bacteriophage. The chromosome of the α‐proteobacterium, Caulobacter crescentus, encodes eight ParE/RelE‐superfamily toxins that are organized into operons with their cognate antitoxins. A systematic genetic analysis of these parDE and relBE TA operons demonstrates that seven encode functional toxins. The one exception highlights an example of a non‐functional toxin pseudogene. Chromosomally encoded ParD and RelB proteins function as antitoxins, inhibiting their adjacently encoded ParE and RelE toxins. However, these antitoxins do not functionally complement each other, even when overexpressed. Transcription of these paralogous TA systems is differentially regulated under distinct environmental conditions. These data support a model in which multiple TA paralogs encoded by a single bacterial chromosome form independent functional units with insulated protein–protein interactions. Further characterization of the parDE1 system at the single‐cell level reveals that ParE1 toxin functions to inhibit cell division but not cell growth; residues at the C‐terminus of ParE1 are critical for its stability and toxicity. While continuous ParE1 overexpression results in a substantial loss in cell viability at the population level, a fraction of cells escape toxicity, providing evidence that ParE1 toxicity is not uniform within clonal cell populations.
The ties that bind: perceived social support, stress, and IBS in severely affected patients
Authors: Lackner J. M. , Brasel A. M. , Quigley B M. , Keefer L. , Krasner S. S. , Powell C. , Katz L. A. , Sitrin M. D.
Published: May 2010

Background  This study assessed the association between social support and the severity of irritable bowel syndrome (IBS) symptoms in a sample of severely affected IBS patients recruited to an NIH‐funded clinical trial. In addition, we examined if the effects of social support on IBS pain are mediated through the effects on stress.

Methods  Subjects were 105 Rome II diagnosed IBS patients (F = 85%) who completed seven questionnaires which were collected as part of a pretreatment baseline assessment.

Key Results  Partial correlations were conducted to clarify the relationships between social support and clinically relevant variables with baseline levels of psychopathology, holding constant number of comorbid medical diseases, age, gender, marital status, ethnicity, and education. Analyses indicated that social support was inversely related to IBS symptom severity. Social support was positively related with less severe pain. A similar pattern of data was found for perceived stress but not quality of life impairment. Regression analyses examined if the effects of social support on pain are mediated by stress. The effects of social support on bodily pain were mediated by stress such that the greater the social support the less stress and the less pain. This effect did not hold for symptom severity, quality of life, or psychological distress.

Conclusions & Inferences  This study links the perceived adequacy of social support to the global severity of symptoms of IBS and its cardinal symptom (pain). It also suggests that the mechanism by which social support alleviates pain is through a reduction in stress levels.
Moonlight and the Migratory Orientation of Savannah Sparrows (Passerculus sandwichensis)
Authors: Moore Frank R.
Published: May 2010

Migratory birds might respond to moonlight in at least four ways: (1) a geographical reference for selecting a compass direction, (2) a celestial ‘landmark’ to facilitate maintenance of a preferred heading, (3) a stimulus that distracts migrants and introduces error in compass orientation, or (4) a source of illumination that facilitates nocturnal flight. This study examines the response of migratory savannah sparrows (Passerculus sandwichensis) to moonlight during controlled tests in orientation cages. I found no evidence that savannah sparrows use a lunar compass to select a direction. If savannah sparrows do use the moon as a ‘landmark’ to maintain a direction selected with reference to a different cue, I expected birds to be better oriented on overcast nights when the moon is present than they are when the moon is absent. The results suggest otherwise. Usually, savannah sparrows respond phototactically to the moon by directing their cage activity toward or at a constant angle with respect to the moon's azimuth. Interestingly, the migrant's response to moonlight depended on whether the bird viewed the setting sun earlier that evening.