bluuebunny/crossref_metadata_2025 huggingface dataset:
86 VIABILITY AND FERTILITY OF RAM SEMEN CRYOPRESERVED WITH DIMETHYL ACETAMIDE AND TREHALOSE
Authors: Bittencourt R. F. , Oba E. , Ribeiro Filho A. de L. , Vasconcelos M. F. , Biscarde C. E. A. , Almeida D. C. , Silveira V. F.
Published: August 2010

Eleven semen samples from Santa Ines breed rams were cryopreserved with the objective of studying the effect of the dimethyl acetamide (DMA, 3%) or glycerol (GL, 6%) in an extender (Tris) with or without trehalose (TRE, 100 mOsmol) on post-thaw sperm viability and fertility. After the evaluation, the semen was diluted in extenders (TRIS+GL6%; TRIS+GL6%+TRE; TRIS+DMA3%; Tris+DMA3%+TRE) and then frozen. The post-thaw kinetic sperm parameters evaluated were the percentage of total motility (TM) and rapid spermatozoa (RAP), track speed (VCL), lateral amplitude of head (ALH), and linearity (LIN). The sperm viability was verified using the fluorescence markers propidium iodide (PMI, plasmatic membrane integrity), JC-1 (MMP, mitochondrial membrane potential), and FITC-PSA (ACI, acrosomal integrity). The spectrum of DNA sperm damage was assessed by single cell gel electrophoresis (Comet assay), and the measurements performed were tail moment (TMOM) and tail intensity (TINT; Comet Assay II software, Perceptive Instruments, Haverhill, UK). The sheep estrus synchronization protocol (n = 170) was 12 days of sponge with 60 mg of medroxyprogesterone acetate and 300 UI of eCG i.m. at sponge withdrawal. The ewes were inseminated on 57.5 h after sponge removal with 150 × 106 of live spermatozoa. The conception rates (CR) were verified for ultrasonography 45 days after insemination. The statistical analysis was performed using SAS software (SAS Institute Inc., Cary, NC, USA). The means (±SEM) of in vitro post-thaw sperm parameters and fertility rates are shown in Table 1. The greatest (P < 0.05) rates of TM and RAP for the GLY groups were observed in relation to the DMA ones. It was also verified the inferiority (P < 0.05) in the VCL and ALH for the trehalose extenders in both groups with GLY and DMA. The values (%) of TINT and TMOM were 14.9 and 03.9, 15.8 and 03.8, 14.7 and 02.9, and 16.6 and 04.4, respectively, for TRIS+GL6%, TRIS+GL6%+TRE, TRIS+DMA3%, and TRIS+DMA3%+TRE (P > 0.05). The integrity rates of all sperm membranes were superior for TRIS+GL6% in relation to TRIS+DMA3%. In conclusion, the DMA had apoor cryoprotective effect for ram semen. The addition of trehalose promoted negative effect on sperm kinetics, although it improved levels of PMM post-thawing in the DMA extender. The conception rate did not present a difference (P > 0.05) among the groups.

Table 1.
99 SURVIVAL RATE AND IN VITRO DEVELOPMENT OF IN VIVO-PRODUCED AND CRYOPRESERVED DOG EMBRYOS
Authors: Luz M. R. , Holanda C. C. , Pereira J. J. , Teixeira N. S. , Vantini R. , Freitas P. M. C. , Salgado A. E. P. , Oliveira S. B. , Guaitolini C. R. F. , Santos M. C.
Published: August 2010

Dogs have been used as an experimental model for human genetic diseases and for research applied to endangered Canidae. Moreover, application of assisted reproductive technologies (ART) by dog breeders is increasing. The aim of this study was to evaluate the survival rate and in vitro development of in vivo-produced and cryopreserved dog embryos. Seven cross-bred bitches were submitted to ovariohysterectomy 12 days after first mating or artificial insemination, and embryos were recovered by uterine horn flushing with 30 mL of PBS/horn (Nutricell, Campinas, SP, Brazil). Grade 1 and 2 morulae (MO; n = 7; IETS) and blastocysts (BL; n = 14) were frozen. For freezing, embryos were immersed in glycerol 10% (GLY; Nutricell, Campinas, SP, Brazil) or ethylene glycol 3,0 M (EG; Nutricell, Campinas, SP, Brazil) for 10 min. Straws were placed in the machine at -7.0°C (TK 3000, Uberaba, MG, Brazil), and equilibrated for 2 min. Seeding was performed at -7.0°C, and another equilibrium period of 2 min was performed. A cooling rate of -0.5°C/min until -32.0°C was used. Embryos were stored in N2L until thawing. Prior to in vitro culture, embryos were removed from N2L, kept at room temperature for 10 s, and put in a water bath at 25°C for 20 s. Embryos frozen in GLY were washed for 5 min in each thawing solution for cryoprotectant removal (0.6 M sacarose + glycerol 5%; 0.6 M sacarose + glycerol 2.5% and 0.6 M sacarose; Nutricell, Campinas, SP, Brazil). After that, embryos were washed 10 times in holding solution (Holding Plus, Bioniche, Pullman, WA, USA). Embryos frozen in EG were kept at room temperature for 10 s, put in a water bath at 25°C for 20 s, and were directly washed 10 times in holding solution. Comparison among groups was performed by ANOVA. After thawing, 9/11 (81.8%) embryos frozen in EG had rupture of zona pelucidae and 2/11 (18.2%) were intact, whereas 9/10 (90.0%) embryos frozen in GLY were intact (P < 0.05). All intact embryos (n = 11) were morphologically normal and were transferred to SOF medium (Nutricell, Campinas, SP, Brazil), cultured for 168 h at 38.3°C, and evaluated at 24-h intervals. After the last evaluation, for both cryoprotectants, hatching rate was 0.0%, but all embryos were morphologically normal. The results of this study suggest that dog embryos have a high in vitro survival rate in standard protocols used for mammalian embryo cryopreservation when glycerol 10% is used as cryoprotectant and that a long period of culture (possibly more than 10 days) is required for dog embryo hatching. On the other hand, this long period for in vitro hatching may reflect the delayed hatching and implantation that occurs in vivo in this species.

Financial by FAPES/MCT/CNPq/CT-INFRA n. 19/2006; Processo n. 36600737/2007, ES- Brasil.
Structure and function of the polymerase core of TRAMP, a RNA surveillance complex
Authors: Hamill Stephanie , Wolin Sandra L. , Reinisch Karin M.
Published: August 2010

The Trf4p/Air2p/Mtr4p polyadenylation (TRAMP) complex recognizes aberrant RNAs in
Saccharomyces cerevisiae
and targets them for degradation. A TRAMP subcomplex consisting of a noncanonical poly(A) RNA polymerase in the Pol ß superfamily of nucleotidyl transferases, Trf4p, and a zinc knuckle protein, Air2p, mediates initial substrate recognition. Trf4p and related eukaryotic poly(A) and poly(U) polymerases differ from other characterized enzymes in the Pol ß superfamily both in sequence and in the lack of recognizable nucleic acid binding motifs. Here we report, at 2.7-Å resolution, the structure of Trf4p in complex with a fragment of Air2p comprising two zinc knuckle motifs. Trf4p consists of a catalytic and central domain similar in fold to those of other noncanonical Pol β RNA polymerases, and the two zinc knuckle motifs of Air2p interact with the Trf4p central domain. The interaction surface on Trf4p is highly conserved across eukaryotes, providing evidence that the Trf4p/Air2p complex is conserved in higher eukaryotes as well as in yeast and that the TRAMP complex may also function in RNA surveillance in higher eukaryotes. We show that Air2p, and in particular sequences encompassing a zinc knuckle motif near its N terminus, modulate Trf4p activity, and we present data supporting a role for this zinc knuckle in RNA binding. Finally, we show that the RNA 3′ end plays a role in substrate recognition.
Anisotropy of wave propagation in the heart can be modeled by a Riemannian electrophysiological metric
Authors: Young Robert J. , Panfilov Alexander V.
Published: August 2010

It is well established that wave propagation in the heart is anisotropic and that the ratio of velocities in the three principal directions may be as large as
v

f

∶
v

s

∶
v

n

 ≈ 4(fibers)∶2(sheets)∶1(normal). We develop an alternative view of the heart based on this fact by considering it as a non-Euclidean manifold with an electrophysiological(
el
-) metric based on wave velocity. This metric is more natural than the Euclidean metric for some applications, because
el
-distances directly encode wave propagation. We develop a model of wave propagation based on this metric; this model ignores higher-order effects like the curvature of wavefronts and the effect of the boundary, but still gives good predictions of local activation times and replicates many of the observed features of isochrones. We characterize this model for the important case of the rotational orthotropic anisotropy seen in cardiac tissue and perform numerical simulations for a slab of cardiac tissue with rotational orthotropic anisotropy and for a model of the ventricles based on diffusion tensor MRI scans of the canine heart. Even though the metric has many slow directions, we show that the rotation of the fibers leads to fast global activation. In the diffusion tensor MRI-based model, with principal velocities 0.25∶05∶1 m/s, we find examples of wavefronts that eventually reach speeds up to 0.9 m/s and average velocities of 0.7 m/s. We believe that development of this non-Euclidean approach to cardiac anatomy and electrophysiology could become an important tool for the characterization of the normal and abnormal electrophysiological activity of the heart.
Evidence of intense ongoing endemic transmission of hepatitis C virus in Egypt
Authors: Miller F. DeWolfe , Abu-Raddad Laith J.
Published: August 2010

Egypt has the highest prevalence of antibodies to hepatitis C virus (HCV) in the world, estimated nationally at 14.7%. An estimated 9.8% are chronically infected. Numerous HCV prevalence studies in Egypt have published various estimates from different Egyptian communities, suggesting that Egypt, relative to the other nations of the world, might be experiencing intense ongoing HCV transmission. More importantly, a new national study provided an opportunity to apply established epidemiologic models to estimate incidence. Validated mathematical models for estimating incidence from age-specific prevalence were used. All previous prevalence studies of HCV in Egypt were reviewed and used to estimate incidence provided that there was sufficient age-specific data required by the models. All reports of anti-HCV antibody prevalence were much higher than any single other national estimate. Age was the strongest and most consistently associated factor to HCV prevalence and HCV RNA positivity. It was not possible to establish a prior reference point for HCV prevalence or incidence to compare with the 2009 incidence estimates. The modeled incidence from the national study and collectively from the modeled incidence from the previous community studies was 6.9/1,000 [95% confidence interval (CI), 5.5–7.4] per person per year and 6.6/1,000 (95% CI, 5.1–7.0) per person per year, respectively. Projected to the age structure of the Egyptian population, more than 500,000 new HCV infections per year were estimated. Iatrogenic transmission is the most likely, underlining exposure to the ongoing transmission. The study demonstrates the urgency to reduce HCV transmission in Egypt.
Hypothesis-driven structural connectivity analysis supports network over hierarchical model of brain architecture
Authors: Thompson Richard H. , Swanson Larry W.
Published: August 2010

The brain is usually described as hierarchically organized, although an alternative network model has been proposed. To help distinguish between these two fundamentally different structure-function hypotheses, we developed an experimental circuit-tracing strategy that can be applied to any starting point in the nervous system and then systematically expanded, and applied it to a previously obscure dorsomedial corner of the nucleus accumbens identified functionally as a “hedonic hot spot.” A highly topographically organized set of connections involving expected and unexpected gray matter regions was identified that prominently features regions associated with appetite, stress, and clinical depression. These connections are arranged as a longitudinal series of circuits (closed loops). Thus, the results do not support a rigidly hierarchical model of nervous system organization but instead indicate a network model of organization. In principle, the double-coinjection circuit tracing strategy can be applied systematically to the rest of the nervous system to establish the architecture of the global structural wiring diagram, and its abstraction, the connectome.
AIDS-protective HLA-B*27/B*57 and chimpanzee MHC class I molecules target analogous conserved areas of HIV-1/SIVcpz
Authors: de Groot Natasja G. , Heijmans Corrine M. C. , Zoet Yvonne M. , de Ru Arnoud H. , Verreck Frank A. , van Veelen Peter A. , Drijfhout Jan W. , Doxiadis Gaby G. M. , Remarque Edmond J. , Doxiadis Ilias I. N. , van Rood Jon J. , Koning Frits , Bontrop Ronald E.
Published: August 2010

In the absence of treatment, most HIV-1-infected humans develop AIDS. However, a minority are long-term nonprogressors, and resistance is associated with the presence of particular HLA-B*27/B*57 molecules. In contrast, most HIV-1-infected chimpanzees do not contract AIDS. In comparison with humans, chimpanzees experienced an ancient selective sweep affecting the MHC class I repertoire. We have determined the peptide-binding properties of frequent chimpanzee MHC class I molecules, and show that, like HLA-B*27/B*57, they target similar conserved areas of HIV-1/SIVcpz. In addition, many animals appear to possess multiple molecules targeting various conserved areas of the HIV-1/SIVcpzGag protein, a quantitative aspect of the immune response that may further minimize the chance of viral escape. The functional characteristics of the contemporary chimpanzee MHC repertoire suggest that the selective sweep was caused by a lentiviral pandemic.
Molecular basis for the resistance of an insect chymotrypsin to a potato type II proteinase inhibitor
Authors: Dunse K. M. , Kaas Q. , Guarino R. F. , Barton P. A. , Craik D. J. , Anderson M. A.
Published: August 2010

Plants produce a variety of proteinase inhibitors (PIs) that have a major function in defense against insect herbivores. In turn, insects have developed strategies to minimize the effect of dietary PIs on digestion. We have discovered that
Helicoverpa
larvae that survive consumption of a multidomain serine PI from
Nicotiana alata
(NaPI) contain high levels of a chymotrypsin that is not inhibited by NaPI. Here we describe the isolation of this NaPI-resistant chymotrypsin and an NaPI-susceptible chymotrypsin from
Helicoverpa
larvae, together with their corresponding cDNAs. We investigated the mechanism of resistance by mutating selected positions of the NaPI-susceptible chymotrypsin using the corresponding amino acids of the NaPI-resistant chymotrypsin. Four critical residues that conferred resistance to NaPI were identified. Molecular modeling revealed that a Phe→Leu substitution at position 37 in the chymotrypsin results in the loss of important binding contacts with NaPI. Identification of the molecular mechanisms that contribute to PI resistance in insect digestive proteases will enable us to develop better inhibitors for the control of lepidopteran species that are major agricultural pests worldwide.
Small-molecule inducers of insulin expression in pancreatic α-cells
Authors: Fomina-Yadlin Dina , Kubicek Stefan , Walpita Deepika , Dančik Vlado , Hecksher-Sørensen Jacob , Bittker Joshua A. , Sharifnia Tanaz , Shamji Alykhan , Clemons Paul A. , Wagner Bridget K. , Schreiber Stuart L.
Published: August 2010

High-content screening for small-molecule inducers of insulin expression identified the compound BRD7389, which caused α-cells to adopt several morphological and gene expression features of a β-cell state. Assay-performance profile analysis suggests kinase inhibition as a mechanism of action, and we show that biochemical and cellular inhibition of the RSK kinase family by BRD7389 is likely related to its ability induce a β-cell-like state. BRD7389 also increases the endocrine cell content and function of donor human pancreatic islets in culture.
Conformational dynamics of bacteriophage T7 DNA polymerase and its processivity factor,
Escherichia coli
thioredoxin
Authors: Akabayov Barak , Akabayov Sabine R. , Lee Seung-Joo , Tabor Stanley , Kulczyk Arkadiusz W. , Richardson Charles C.
Published: August 2010

Gene 5 of bacteriophage T7 encodes a DNA polymerase (gp5) responsible for the replication of the phage DNA. Gp5 polymerizes nucleotides with low processivity, dissociating after the incorporation of 1 to 50 nucleotides. Thioredoxin (trx) of
Escherichia coli
binds tightly (Kd = 5 nM) to a unique segment in the thumb subdomain of gp5 and increases processivity. We have probed the molecular basis for the increase in processivity. A single-molecule experiment reveals differences in rates of enzymatic activity and processivity between gp5 and gp5/trx. Small angle X-ray scattering studies combined with nuclease footprinting reveal two conformations of gp5, one in the free state and one upon binding to trx. Comparative analysis of the DNA binding clefts of DNA polymerases and DNA binding proteins show that the binding surface contains more hydrophobic residues than other DNA binding proteins. The balanced composition between hydrophobic and charged residues of the binding site allows for efficient sliding of gp5/trx on the DNA. We propose a model for trx-induced conformational changes in gp5 that enhance the processivity by increasing the interaction of gp5 with DNA.
Epidemiology of Helicobacter pylori infection.
Authors: Malaty H. M.
Published: August 2010

This chapter discusses the relationship between Helicobacter pylori infection and associated diseases (chronic gastritis, peptic ulcer disease and gastric adenocarcinoma) and describes the route of H. pylori transmission. The geographical distribution and changing trends of the prevalence of H. pylori infection, the changes in the environment and human behaviour that influence the prevalence of H. pylori infection, and the decreasing prevalence of infection-induced ulcers in western countries are reviewed.
Innate immune initiators and effectors in Helicobacter pylori infection.
Authors: Kaparakis M. , Allison C. C. , Hutton M. L. , Ferrero R. L.
Published: August 2010

This chapter discusses the innate immune responses generated by the host in an attempt to control Helicobacter pylori infection. The roles of host pathogen-recognition molecules in the identification of H. pylori within the gastric mucosa and the downstream signalling events that these molecules trigger are described. Some of the key H. pylori products that facilitate the initiation of inflammation are identified. How these processes result in the production of proinflammatory cytokines, ultimately leading to the development of gastritis within the host, are discussed.
Improving college students’ evaluation of text learning using idea-unit standards
Authors: Dunlosky John , Hartwig Marissa K. , Rawson Katherine A. , Lipko Amanda R.
Published: August 2010

When recalling key definitions from class materials, college students are often overconfident in the quality of their responses. Even with commission errors, they often judge that their response is entirely or partially correct. To further understand this overconfidence, we investigated whether idea-unit judgements would reduce overconfidence (Experiments 1 and 2) and whether students inflated their scores because they believed that they knew answers but just responded incorrectly (Experiment 2). College students studied key-term definitions and later attempted to recall each definition when given the key term (e.g., What is the availability heuristic?). All students judged the quality of their recall, but some were given a full-definition standard to use, whereas other students first judged whether their response included each of the individual ideas within the corresponding correct answer. In Experiment 1, making these idea-unit judgements reduced overconfidence for commission errors. In Experiment 2, some students were given the correct definitions and graded other students’ responses, and some students generated idea units themselves before judging their responses. Students were overconfident even when they graded other students’ responses, and, as important, self-generated idea units for each definition also reduced overconfidence in commission errors. Thus, overconfidence appears to result from difficulties in evaluating the quality of recall responses, and such overconfidence can be reduced by using idea-unit judgements.
Central nervous system (CNS)–resident natural killer cells suppress Th17 responses and CNS autoimmune pathology
Authors: Hao Junwei , Liu Ruolan , Piao Wenhua , Zhou Qinghua , Vollmer Timothy L. , Campagnolo Denise I. , Xiang Rong , La Cava Antonio , Van Kaer Luc , Shi Fu-Dong
Published: August 2010

Natural killer (NK) cells of the innate immune system can profoundly impact the development of adaptive immune responses. Inflammatory and autoimmune responses in anatomical locations such as the central nervous system (CNS) differ substantially from those found in peripheral organs. We show in a mouse model of multiple sclerosis that NK cell enrichment results in disease amelioration, whereas selective blockade of NK cell homing to the CNS results in disease exacerbation. Importantly, the effects of NK cells on CNS pathology were dependent on the activity of CNS-resident, but not peripheral, NK cells. This activity of CNS-resident NK cells involved interactions with microglia and suppression of myelin-reactive Th17 cells. Our studies suggest an organ-specific activity of NK cells on the magnitude of CNS inflammation, providing potential new targets for therapeutic intervention.
High current fast switching n-ZnO/p-Si diode
Authors: Choi Young , Lee Kimoon , Park C H , Lee Kwang H , Nam Jae-Woo , Sung Myung M , Lee Kyu Min , Sohn Hyun Chul , Im Seongil
Published: August 2010

The authors report on the high current density n-ZnO/p-Si heterojunction diode that has been fabricated by atomic layer deposition (ALD) of 70 nm thin ZnO on a p-Si substrate. While the diode was formed at four different ALD temperatures of 80, 100, 150 and 200 °C, the 100 °C processed diode showed an optimal behaviour of an on–off ratio over 3.3 × 103 and a high forward current density, ∼300 A cm−2 at 3 V. Although the highest film conductance appeared from the 200 °C deposited ZnO layer, nanometre thin SiO
x
was also revealed at the ZnO/p-Si interface; it might cause a high reverse leakage current level. Our high current density diode also demonstrates a fast switching performance without any reverse recovery delay, which is often observed at a usual Si p–n diode.
The population distribution of argon atoms in Paschen 1s levels in an inductively coupled plasma
Authors: Li Jiang , Zhu Xi-Ming , Pu Yi-Kang
Published: August 2010

The population distribution of argon atoms in Paschen 1s levels in inductively coupled plasmas is investigated using a collisional–radiative model and the optical emission spectroscopy method. The modelling results of population densities are in good agreement with the experimental ones. According to this model, the population distribution of 1s levels is affected mainly by the electron impact transfer and the resonance radiation processes. As a result, a simple relationship on the population ratio of 1s4 and 1s5 is obtained. From this relationship, three kinetic regimes with different electron densities and discharge pressures are identified, which can be used to characterize the population distribution of argon 1s levels in inductively coupled plasmas.
Dwell time dependent morphological transition and sputtering yield of ion sputtered Sn
Authors: Qian H X , Zhou W , Zeng X R
Published: August 2010

Self-organized nano-scale patterns may appear on a wide variety of materials irradiated with an ion beam. Good manipulation of these structures is important for application in nanostructure fabrication. In this paper, dwell time has been demonstrated to be able to control the ripple formation and sputtering yield on Sn surface. Ripples with a wavelength of 1.7 µm were observed for a dwell time in the range 3–20 µs, whereas much finer ripples with a wavelength of 540 nm and a different orientation were observed for a shorter dwell time in the range 0.1–2 µs. The sputtering yield increases with dwell time significantly. The results provide a new basis for further steps in the theoretical description of morphology evolution during ion beam sputtering.
The misuse of terms in scientific literature
Authors: Marabotti Anna , Facchiano Angelo
Published: August 2010

Contact:  angelo.facchiano@isa.cnr.it
MASiVE: Mapping and Analysis of SireVirus Elements in plant genome sequences
Authors: Darzentas N. , Bousios A. , Apostolidou V. , Tsaftaris A. S.
Published: August 2010

Summary: We present MASiVE, an expertly built tool for the large-scale, yet sensitive and highly accurate, discovery, preliminary analysis and insertion age estimation of intact Sirevirus LTR-retrotransposons in plant genomic sequences. Validation was based on the recently available and annotated large maize chromosome one. Results show a considerable improvement in the annotation of Sireviruses, and support our approach as an important addition to the bioinformatics toolbox of plant biologists.

Availability: PERL source code and essential files are available online at http://bat.ina.certh.gr/tools/masive/. The freely available Vmatch, LTRharvest, Wise2, and MAFFT algorithms are required.

Contact:  ndarz@certh.gr

Supplementary information:  Supplementary data are available at Bioinformatics online.
Stem Rot of Branched Broomrape (Orobanche ramosa) Caused by Sclerotium rolfsii in Chile
Authors: Galdames R. , Diaz J.
Published: August 2010

Branched broomrape is a holoparasitic weed present in some areas of central and southern Chile (33°S to 38°S), which is often found parasitizing tomato and tobacco crops. During an extensive survey conducted in different tomato-growing areas during the summer of 2010, branched broomrape plants with stem rot symptoms were detected in a commercial tomato crop located in the central zone (34°14′S, 71°1′W). Rotten stems were observed with white mycelia and approximately 1-mm-diameter spherical sclerotia on affected tissue below the soil surface. Parasitized tomato plants showed no symptoms. Sclerotia were taken directly from affected stems with a dissecting needle under a stereoscopic microscope in a flow chamber and placed on potato dextrose agar (PDA) medium. Germinating sclerotia consistently produced colonies similar to Sclerotium rolfsii with new sclerotia formed within 6 to 7 days. Mycelia produced hyphal clamp connections under the light microscope (2). DNA was extracted from one fungal culture. The ITS1 region, 5.8S rRNA gene, and the ITS2 region of the nuclear-encoded ribosomal RNA gene were amplified with primers ITS1 and ITS4 (4). The sequence was deposited in GenBank (Accession No. HM222638) and showed ≥99% identity values with sequences of similar regions from Athelia rolfsii (anamorph S. rolfsii; Accession Nos. AB075304, DQ0595578, AF499018, and AB075305). Different pathogenicity tests were performed. Inoculum was prepared by placing mycelia plugs from a PDA-grown, 6-day-old colony in a flask with sterilized wheat seeds and incubated for 2 weeks. Three Orobanche plants, each one with 10 to 15 shoots at different aerial stages (starting emergence, flowering, and formed capsules) were gently planted separately in 35-cm pots. Inoculum (10 g per pot) was placed in the soil surrounding the plants. One pot was used as a control. Forty-five-day-old tomato and tobacco plants were additionally inoculated by a similar procedure. After 12 days, inoculated Orobanche plants showed reduced vigor and stem decay. After 9 days, tomato and tobacco plants showed wilt. In all cases, the fungus was reisolated on PDA from all inoculated plants. To our knowledge, this is the first report of S. rolfsii on O. ramosa (1). A previous report of S. rolfsii parasitizing O. cernua has also been made (3). The high susceptibility of tomato and tobacco plants to this isolate of S. rolfsii precludes the use of this pathogen as a biological control agent against broomrape.

References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved May 14, 2010, from http://nt.ars-grin.gov/fungaldatabases . (2) Z. K. Punja and A. Damiani. Mycologia 88:694, 1996. (3) C. A. Raju et al. Phytoparasitica 23:235, 1995. (4) T. J. White et al. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, New York, 1990.
First Report of Southern Blight of Okra (Abelmoschus esculentus) Caused by Sclerotium rolfsii in Côte d'Ivoire
Authors: Koné D. , Mohamed D. , Soro S. , Bolou Bi B. A. , Kouadio Y. J. , Ji P.
Published: August 2010

Southern blight caused by Sclerotium rolfsii Sacc. was observed on okra (Abelmoschus esculentus) cv. Clemson Spineless in January 2010 during a survey of vegetable fields in Rubbino, Côte d'Ivoire, which is one of the most important areas for vegetable production. Plants exhibited symptoms of a dark brown lesion on the stems near the soil line. Upper roots became light to dark brown, the lower leaves wilted, turned yellow, and a white mat of fungal mycelium developed on the stem lesion. Numerous, white, spherical sclerotia formed on the infected stem and on soil surfaces around the infected plants. Sclerotia (0.5 to 1.2 mm in diameter) later turned tan to dark brown and the entire plant wilted. Eleven fields, with a total of approximately 26 ha surveyed, were affected and disease incidence reached approximately 15% in the fields. Sclerotia were collected and the fungus cultured on acidified potato dextrose agar (PDA) plates. The fungus grew rapidly on PDA and the hyphae at the edge of the colonies were large straight cells with one or more clamp connections at each septum. Secondary and tertiary hyphae were slender and lacked clamp connections. Whitish sclerotia were observed on the mycelial mats 5 to 7 days after incubation, which later turned tan to brown when mature. The fungus was identified as S. rolfsii based on the characteristics of mycelia and sclerotia (1). Sclerotia produced on PDA were used to inoculate okra seedlings under greenhouse conditions at rates of 10, 20, and 30 sclerotia per plant. Sclerotia were placed just under the soil surface around the root of 4-week-old seedlings of okra cv. Clemson Spineless. Symptoms identical to those on field samples developed on all inoculated plants. S. rolfsii was reisolated from diseased plants and the identity was confirmed. S. rolfsii has previously been reported as causing disease in Côte d'Ivoire (2). To our knowledge, this is the first report of southern blight caused by S. rolfsii on okra in this country. Okra is an important vegetable crop in Côte d'Ivoire and therefore the occurrence of southern blight and susceptibility of okra cultivars to this disease needs to be taken into account in okra production.

References: (1) Z. K. Punja and A. Damiani. Mycologia 88:694, 1996. (2) R. Resplandy et al. Ann. Epiphyt. 1:1, 1954.
Mancozeb: Past, Present, and Future
Authors: Gullino Maria Lodovica , Tinivella Federico , Garibaldi Angelo , Kemmitt Gregory M. , Bacci Leonardo , Sheppard Brian
Published: August 2010

This feature article reviews the broad-spectrum fungicide mancozeb. Introduced in 1962, it still plays a significant role in the world fungicide market. Mancozeb possesses a number of key attributes that have contributed toward its development into a globally important tool in modern chemical-based plant disease management. These attributes are discussed from the perspective of both public and private research.
Direct Polymerase Chain Reaction-Based Detection of Cercospora beticola in Field Soils
Authors: Lartey R. T. , Caesar-TonThat T. C. , Lenssen A. W. , Eckhoff J. , Hanson S. L. , Evans R. G.
Published: August 2010

Cercospora beticola, the causal agent of Cercospora leaf spot of sugar beet, survives as pseudostromata in infected sugar beet residues in the soil. Under optimal conditions, overwintering propagules germinate and produce conidia that are dispersed as primary inoculum to initiate infection in sugar beet. We developed a polymerase chain reaction (PCR) technique for rapid detection of C. beticola in field soils. Total DNA was first isolated from soil amended with C. beticola culture using the PowerSoil DNA Kit. The purified DNA was subjected to PCR in Extract-N-Amp PCR mix with CBACTIN primers over 35 cycles. The amplified products were resolved and compared by electrophoresis in 1% agarose gels. The PCR fragment size of C. beticola from the amended field soil correlated in size with the amplicon from control C. beticola culture DNA extract. Additionally, sample soils were collected from sugar beet fields near Sidney, MT and Foxholm, ND. Total DNA was extracted from the samples and subjected to PCR and resolved as previously described. The amplicons were purified from the gels and subjected to BigDye Terminator Cycle sequencing. All sequences from field soils samples, C. beticola-amended field soil, and pure culture were compared by alignment with a C. beticola actin gene sequence from GenBank. The result of the alignment confirmed the amplicons as products from C. beticola. Rapid screening for the presence of C. beticola in the soil by PCR will improve research capabilities in biological control, disease forecasting, and management of this very important sugar beet pathogen.
Effect of Sclerotium Density and Irrigation on Disease Incidence and on Efficacy ofConiothyrium minitans in Suppressing Lettuce Drop Caused bySclerotinia sclerotiorum
Authors: Chitrampalam P. , Turini T. A. , Matheron M. E. , Pryor B. M.
Published: August 2010

Field experiments were conducted over 2 years in Yuma, AZ, and Holtville, CA, to establish the relationship between soil sclerotium density of Sclerotinia sclerotiorum and the incidence of lettuce drop on different lettuce (Lactuca sativa) types under different irrigation systems, and to determine the efficacy of the biocontrol agent Coniothyrium minitans (Contans) against S. sclerotiorum on crisphead lettuce at varied sclerotium densities under different irrigation systems. There was no significant interaction of irrigation (overhead sprinkler versus furrow) with either sclerotium density or with biocontrol treatment. Lettuce drop incidence was lowest in romaine lettuce compared with crisphead or leaf lettuce at all soil sclerotium densities. There was a significant positive correlation between the sclerotial density and the percent disease incidence. Disease incidence in plots infested with 2 sclerotia/m2of bed was not significantly higher than in control plots regardless of lettuce type. However, plots infested with 40 or 100 sclerotia/m2of bed revealed a significantly higher disease incidence over the control in all lettuce types. A single application of Contans at planting significantly reduced the incidence of lettuce drop in all lettuce types even under high disease pressure. There were no significant differences between recommended (2.2 kg/ha) and high (4.4 kg/ha) application rates of Contans or between one or two applications of the product.
First Report of Root and Basal Rot of Angelica acutiloba Caused by Fusarium solani in Taiwan
Authors: Chern L. L. , Feng C. T. , Yu C. H. , Ho W. C.
Published: August 2010

Angelica (Angelica acutiloba (Siebold. & Zucc.) Kitag.) is one of the most important traditional Chinese medicines in Taiwan. The medicinal herb has been mainly imported from China, but cultivation at a commercial scale has also been established in recent years in Hualien County, Taiwan. In September 2008, angelica plants in a field at Liou-shih-dan Mountain displayed symptoms of yellowing, stunting, rotting of roots and basal stem, and wilting. A severe brown discoloration of vascular tissue along the stems of infected plants was observed. One or more Fusarium spp. was consistently isolated from the roots and stems of diseased plants. Isolates R3, R4, and R5 were incubated for 14 days on celery tissues to produce chlamydospores, and 33 g of celery tissue with chlamydospores were mixed with 500 ml of soil per pot as inoculum. One 4-month-old angelica seedling was planted per pot. Three angelica plants were inoculated with each isolate in the first test and nine plants were inoculated with each isolate in the second test. Other seedlings were inoculated with water as checks. Pathogenicity tests were conducted twice. Incidence of diseased plants was 66, 100, and 33% in the first test, and 66, 100, and 44% in the second test for the R3, R4, and R5 isolates, respectively. Symptoms similar to those on the diseased plants in the field were produced, with leaves turning yellow starting 7 days after inoculation and wilt and discoloration of roots 14 days after inoculation. Fusarium spp. also were reisolated from the diseased plants. Genomic DNA was extracted from mycelium with a fungal genomic DNA purification kit, and the internal transcribed spacer (ITS) rDNA region was amplified and sequenced with primers ITS-4 and ITS-5. The sequence of the resulting ~550-bp amplicon was compared with those in GenBank. The ITS sequences of the R3, R4, and R5 isolates shared 98.7, 98.7, and 97.9% similarity with F. solani isolate AF129104 (3), respectively. Phylogenetic analysis also showed that the three isolates were closer to F. solani than to other Fusarium species. Both macroconidia and microconidia of the R4 isolate were produced on potato dextrose agar. Macroconidia were three to five septate and 27.2 to 37.8 × 4.4 to 6.2 μm; microconidia were zero to one septate and 9.3 to 14.7 × 2.9 to 4.8 μm. Chlamydospores produced on celery juice agar were terminal or intercalary, solitary, in pairs or in chains, and 9.3 to 12.1 μm. Morphological characteristics identified the three isolates as F. solani (Martius) Snyder & Hansen according to Fu and Chang (2) and Chung et al. (1), which agrees with the ITS comparison. To our knowledge, this is the first report of root and basal rot caused by F. solani on angelica in Taiwan.

References: (1) W. C. Chung et al. Plant Prot. Bull. 40:177, 1998. (2) C. H. Fu and T. T. Chang. Taiwan J. For. Sci. 14:223, 1999. (3) H. Suga et al. Mycol. Res. 104:1175, 2000.
First Report of the Telial Stage of Japanese Apple Rust on Juniperus chinensis in North America and the Aecial Stage on Malus domestica
Authors: Gregory N. F. , Bischoff J. F. , Dixon L. J. , Ciurlino R.
Published: August 2010

Following a report in April 2009 of the presence of Gymnosporangium yamadae Miyabae ex G. Yamada on crabapple (Malus toringo Siebold) in Wilmington, DE (2), University of Delaware, State of Delaware, and USDA/APHIS PPQ personnel collaborated to confirm and document the pathogen. G. yamadae is the causal agent of Japanese apple rust. The fungus is known from Asia with an aecial state on economically important Malus species and telial state on Juniperus chinensis. During the April 2009 site visit, ornamental J. chinensis were observed near the original crabapples. On May 7, 2009, telial galls were collected from the ornamental J. chinensis at the Wilmington site. The telia were confirmed to be G. yamadae by morphometric analysis and molecular data. The rDNA large subunit (LSU) sequence derived from the collected telial galls (GenBank Accession No. GU058012) was identical to the eight G. yamadae LSU sequences (GenBank Accession Nos. FJ848760–FJ848765, FJ559373, and FJ559375) reported from Korea by Yun et al. (3). Teliospores were 45 to 54 μm long with pedicels that were wide (7.0 to 8.4 μm) along the full length. The G. yamadae telial gall collected from Wilmington, DE was deposited into the U.S. National Fungus Collection (BPI 879273). Leaves of M. domestica on the University of Delaware farm in Newark were confirmed to have Japanese apple rust on Aug 4, 2009. Identification was made on the morphological presence of unique roestelioid aecia with long cornulated peridia that lacerate along the sides. The aecia differ from those of G. juniperi-virginianae, the causal agent of cedar apple rust, which has aecial peridia that fimbriate to the base and are strongly recurved (1). Following release of a USDA Pest Alert, subsequent samples submitted to USDA/APHIS PPQ indicated widespread incidence of the G. yamadae aecial state in the northeast, including Maryland, Maine, New Hampshire, New Jersey, New York, Pennsylvania, and Rhode Island. Japanese apple rust likely went undetected for several years because of similar symptomatology to cedar apple rust. To our knowledge, this is the first report of the telial stage of G. yamadae in North America and the first report of this pathogen on Malus domestica in the United States. Knowledge of the geographic distribution of G. yamadae is of significance because of the actionable regulatory status of the pathogen and its potential impact on ornamental and fruit growers of Malus spp. in the United States.

References: (1) F. D. Kern. A Revised Taxonomic Account of Gymnosporangium. Pennsylvania State University Press, University Park, PA, 1973. (2) H. Y. Yun et al. Plant Dis. 93:430, 2009. (3) H. Y. Yun et al. Mycologia 101:790, 2009.
First Report of Phytophthora ramorum Infecting California Red Fir in California
Authors: Chastagner G. A. , Riley K. L.
Published: August 2010

In May 2005, branches originating from five separate whorls below the terminal on a single California red fir (Abies magnifica) in a mixed grand fir (Abies grandis) and Douglas-fir (Pseudotsuga menziesii) Christmas tree plantation near Los Gatos, CA displayed wilting and dieback of new shoot growth. Brown dieback, delineated by needle loss, extended 6 to 8 cm into 1-year-old and sometimes 2-year-old growth. The ~7-year-old, 1-m tall tree was located near the edge of the plantation, beneath an overstory of California bay laurel (Umbellularia californica) trees that were infected with Phytophthora ramorum. Isolations from dieback margins onto corn meal agar amended with ampicillin, rifamycin, and pimaricin (CARP) yielded hyphae and large, dark brown chlamydospores that were morphologically consistent with P. ramorum (1). Microsatellite analysis confirmed that isolates were of the NA1 lineage of P. ramorum. Isolates were deposited in the Washington State University Puyallup Phytophthora Master Collection. Dormant bareroot California red fir seedlings were obtained from the USDA Forest Service Placerville Nursery (Camino, CA) in February 2006 and planted in SC-10 super cell cones (Stuewe & Sons, Inc., Tangent, OR) in a standard greenhouse potting mix. Seedlings (average height 11 cm) were then forced to initiate bud break and new shoot elongation (0.5 to 1.5 cm) in a greenhouse at 21°C. Eight unwounded seedlings were inoculated with a zoospore suspension (4.185 × 105 zoospores/ml of sterile water) produced from 3- to 4-week-old V8 juice agar cultures of isolate WSU#106-0021 using an artist's airbrush powered by Badger Propel canned propellant. Eight control seedlings were sprayed with water alone. Seedlings were placed in plastic tubs with ~2.5 cm of warm water in the bottom to provide humidity. A plastic bag supported by a wire frame was used to cover each tub. Tubs were placed in a biocontainment unit at 15 to 16°C under 24 h of fluorescent light. The plastic was removed after 5 days and seedlings were left under the same conditions. Seven days after inoculation, 25 to 100% (average 68%) of the new shoots on each of the eight inoculated seedlings were wilted and 100% of these seedlings exhibited dark brown dieback into the 1-year-old stems (range of 1.0 to 2.3 cm, average 1.6 cm). Tissues from shoots and dieback edges were plated onto CARP media. All of these attempts resulted in successful isolation of P. ramorum, and cultures exhibited the same hyphal morphology and chlamydospore characteristics when compared with the isolate tested. Control plants did not develop symptoms. This trial completes Koch's postulates to establish California red fir as a host of Phytophthora ramorum. To our knowledge, this site contains the only reported infection of California red fir by P. ramorum. The potential for infection within its native range is unknown.

Reference: (1) S. Werres et al. Mycol. Res. 105:1155, 2001.
First Report of Tomato chlorotic dwarf viroid in Petunia spp. in Slovenia
Authors: Viršček Marn M. , Mavrič Pleško I.
Published: August 2010

In early April 2010, 30 samples of Petunia spp. were taken by phytosanitary inspectors from 22 production sites in Slovenia in the frame of surveying host plants for the presence of Potato spindle tuber viroid (PSTVd). Samples were taken in accordance with the plan of the survey for the year 2010 and were tested for the presence of PSTVd by real-time RT-PCR according to the EPPO protocol (1). At the time of sampling, there were no disease symptoms on the plants. Samples consisted of fully developed leaves collected from as many as five plants. Total RNA was isolated from 50 ± 5 mg of leaf tissue with an RNeasy Plant Mini Kit (Qiagen, Chatsworth, CA). One sample of cv. Surfinia Purple from a production site from the coastal region and another of cv. Surfinia Hot Pink 05 from a production site near Ljubljana, both multiplied through cuttings, were positive by real-time RT-PCR, confirming the presence of PSTVd or Tomato chlorotic dwarf viroid (TCDVd). To identify the viroid, RT-PCR with primer pairs of Shamloul et al. (3) and Di Serio (2) were performed with isolated total RNA of each positive sample. RT-PCR products were obtained only with primer pairs of Shamloul et al. (3). To obtain the full sequence, additional RT-PCR was done for each sample with semi-universal pospiviroid primers Vid-RE/FW (4). RT-PCR products obtained with primer pair of Shamloul et al. (3) and primer pair Vid RE/FW were sequenced (Macrogen, Seoul, Korea). Sequence analysis confirmed the identity of a viroid as TCDVd. Both isolates consisted of 360 nucleotides and were 100% identical to an isolate from tomato deposited in NCBI GenBank under Accession No. AF162131. They showed 98% identity with sequences from petunias (GQ396664, EF582392, EF582393, and DQ859013). The infected Petunia spp. stocks were destroyed. Although the infection of Petunia spp. with TCDVd is symptomless, the infected plants could be a source of infection for tomato and potato. TCDVd infection can cause severe damage on potato and tomato, similar to that caused by infection with PSTVd, to which it is closely related. To our knowledge this is the first finding of TCDVd in Petunia spp. in Slovenia.

References: (1) Anonymous. EPPO Bull. 34:257, 2004. (2) F. Di Serio. J. Plant Pathol. 89:297, 2007. (3) A. M. Shamloul et al. Can. J. Plant Pathol. 19:89, 1997. (4) J. Th. J. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004.
QUALITY AND STORAGE STABILITY OF FRANKFURTERS CONTAINING 15% MECHANICALLY DEBONDED TURKEY MEAT
Authors: FRONING G. W. , ARNOLD R. G. , MANDIGO R. W. , NETH C. E. , HARTUNG T. E.
Published: August 2010

SUMMARY –Fresh and frozen stored mechanically deboned turkey meat was incorporated into frankfurters at the 15% level and compard to red meat frankfurters. Mechanically deboned turkey meat exhibited higher emulsifying capacity than beef but lower than pork on a protein basis. This trend for emulsifying capacity was reversed when reported on a total meat basis. Emulsion stability was not essentially affected by the addition of 15% mechanically deboned turkey meat in red meat frankfurters. Differences flavor tests, preference flavor tests, and TBA values indicated that frankfurters containing 15% mechanically deboned turkey meat were comparable to all red meat frankfurters in flavor stability if fresh deboned poultry meat is used. The use of mechanically deboned poultry meat, which had undergone 90 days of frozen storage, resulted in a significantly inferior product as indicated by flavor evaluation and TBA values. Color evaluation showed slight color fading of all frankfurter treatments during storage. Microbial loads in the three frankfurther treatments showed some increased in total counts during refrigerated storage.