Stunting of Onion in the Columbia Basin of Oregon and Washington Caused by Rhizoctonia spp.

During 2009 and 2010, 45 isolates of Rhizoctonia spp. were recovered from onion bulb crops in the semiarid Columbia Basin of Oregon and Washington, in which patches of severely stunted onion plants developed following rotation with winter cereal cover crops. Characterization of isolates recovered from naturally infested soil and roots was performed by sequence analysis of the ribosomal DNA (rDNA) internal transcribed spacer region, with the majority of isolates (64%) identified as Rhizoctonia solani. In steam-pasteurized field soil, stunting of onion was caused by isolates of R. solani anastamosis groups (AGs) 2-1, 3, 4, and 8, as well as Waitea circinata var. circinata and binucleate Rhizoctonia AG E evaluated at 13 and 8 or 15 and 15°C day and night temperatures, respectively, typical of spring planting conditions in the Columbia Basin. Isolates of R. solani AG 5 as well as binucleate AG A and I were nonpathogenic. The most virulent isolates belonged to AG 8, although an AG 3 and an AG E isolate were also highly virulent. Isolates of AG 2-1 and 3 caused moderate levels of disease, while isolates of AG 4 and W. circinata var. circinata caused low levels of disease. Emergence was reduced by isolates of AG 2-1, 3, and E. When the various AGs were grown at temperatures of 5 to 30°C, the relative growth rate of the Rhizoctonia isolates was not positively correlated with virulence on onion within an AG.

NMR analysis of metabolic responses to extreme conditions of the temperature-dependent coral pathogen Vibrio coralliilyticus.

AIMS: To identify and understand the presence of metabolites responsible for the variation in the metabolic profile of Vibrio coralliilyticus under extreme conditions. METHODS AND RESULTS: Multiple batches of V. coralliilyticus were grown under normal conditions. Four samples in one batch were subjected to extreme conditions via a freeze-thaw cycle during lyophilization. Polar metabolites were extracted using a combination of methanol, water and heat. Nuclear magnetic resonance (NMR)-based metabolic profiles indicated significant differences between the normal and stressed samples. Three compounds identified in the stressed metabolome were maltose, ethanolamine, and the bioplastic-type compound (BTC) 2-butenoic acid, 2-carboxy-1-methylethyl ester. This is the first report of the production of this BTC by V. coralliilyticus. CONCLUSIONS: The presence of maltose and ethanolamine indicates a state of acute nutrient limitation; therefore, we hypothesize that the cell's metabolism turned to its own cell wall, or perhaps neighbouring cells, for sources of carbon and nitrogen. The presence of the BTC also supports the acute nutrient limitation idea because of the parallels with polyhydroxyalkanoate (PHA) production in other gram-negative bacteria, including other Vibrio species. SIGNIFICANCE AND IMPACT OF THE STUDY: Recent metabolomics research on the temperature-dependent coral pathogen V. coralliilyticus has led to the discovery of several compounds produced by the organism as a response to high density, low nutrient conditions. The three metabolites, along with (1) H NMR metabolic fingerprints of the nutrient limited samples, are proposed to serve as metabolic markers for extremely stressful conditions of V. coralliilyticus.

Control of Common Bunt in Organic Wheat.

Common bunt, caused by the seedborne and soilborne pathogens Tilletia caries and T. laevis, has re-emerged as a major disease in organic wheat. In conventional agriculture, common bunt is routinely managed with the use of synthetic chemical seed treatments. For this reason, common bunt is a relatively unimportant disease in conventional agriculture. However, since synthetic chemical inputs are prohibited in organic agriculture, common bunt is a major threat once more in organic wheat and seed production. The challenge today is to manage the disease without the use of chemical seed treatments. This review reports on the management of common bunt under organic farming systems, mainly through host resistance and organic seed treatments. We report the history of screening wheat germplasm for bunt resistance, the search for new sources of resistance, and identification and mapping of bunt resistance genes. Since the pathogen has a gene-for-gene relationship with the host, this review also includes a summary of work on pathogen race identification and virulence patterns of field isolates. Also included are studies on the physiological and molecular basis of host resistance. Alternative seed treatments are discussed, including physical seed treatments, and microbial-based and plant-based treatments acceptable in organic systems. The article concludes with a brief discussion on the current gaps in research on the management of common bunt in organic wheat.

Human GM-CSF: molecular cloning of the complementary DNA and purification of the natural and recombinant proteins.

Clones of complementary DNA encoding the human lymphokine known as granulocyte-macrophage colony-stimulating factor (GM-CSF) were isolated by means of a mammalian cell (monkey COS cell) expression screening system. One of these clones was used to produce recombinant GM-CSF in mammalian cells. The recombinant hematopoietin was similar to the natural product that was purified to apparent homogeneity from medium conditioned by a human T-cell line. The human T-cell GM-CSF was found to be 60 percent homologous with the GM-CSF recently cloned from murine lung messenger RNA.

HDAC6 selective inhibition of melanoma patient T-cells augments anti-tumor characteristics.

BACKGROUND: Therapies targeting anti-tumor T-cell responses have proven successful in the treatment of a variety of malignancies. However, as most patients still fail to respond, approaches to augment immunotherapeutic efficacy are needed. Here, we investigated the ability of histone deacetylase 6 (HDAC6)-selective inhibitors to decrease immunosuppression and enhance immune function of melanoma patient T-cells in ex vivo cultures. METHODS: T-cells were harvested from peripheral blood or tumor biopsies of metastatic melanoma patients and cultured in the presence of pan-, class-specific or class-selective histone deacetylase (HDAC) inhibitors. Changes in cytokine production were evaluated by Luminex and intracellular flow cytometry staining. Expression of surface markers, transcription factors, protein phosphorylation, and cell viability were assessed by flow cytometry. Changes in chromatin structure were determined by ATAC-seq. RESULTS: T-cell viability was impaired with low doses of pan-HDAC inhibitors but not with specific or selective HDAC inhibitors. The HDAC6-selective inhibitors ACY-1215 (ricolinostat) and ACY-241 (citarinostat) decreased Th2 cytokine production (i.e. IL-4, IL-5, IL-6, IL-10 and IL-13). Expansion of peripheral blood T-cells from melanoma patients in the presence of these inhibitors resulted in downregulation of the Th2 transcription factor GATA3, upregulation of the Th1 transcription factor T-BET, accumulation of central memory phenotype T-cells (CD45RA-CD45RO + CD62L + CCR7+), reduced exhaustion-associated phenotypes (i.e. TIM3 + LAG3 + PD1+ and EOMES+PD1+), and enhanced killing in mixed lymphocyte reactions. The frequency, FOXP3 expression, and suppressive function of T regulatory cells (Tregs) were decreased after exposure to ACY-1215 or ACY-241. Higher frequencies of T-cells expressing CD107a + IFNγ+ and central memory markers were observed in melanoma tumor-infiltrating lymphocytes (TIL), which persisted after drug removal and further expansion. After ACY-1215 treatment, increased chromatin accessibility was observed in regions associated with T-cell effector function and memory phenotypes, while condensed chromatin was found in regions encoding the mTOR downstream molecules AKT, SGK1 and S6K. Decreased phosphorylation of these proteins was observed in ACY-1215 and ACY-241-treated T-cells. AKT- and SGK1-specific inhibition recapitulated the increase in central memory frequency and decrease in IL-4 production, respectively, similar to the observed effects of HDAC6-selective inhibition. CONCLUSIONS: HDAC6-selective inhibitors augmented melanoma patient T-cell immune properties, providing a rationale for translational investigation assessing their potential clinical efficacy.

HDAC1,2 inhibition and doxorubicin impair Mre11-dependent DNA repair and DISC to override BCR-ABL1-driven DSB repair in Philadelphia chromosome-positive B-cell precursor acute lymphoblastic leukemia.

Philadelphia chromosome-positive (Ph+) B-cell precursor acute lymphoblastic leukemia (ALL) expressing BCR-ABL1 oncoprotein is a major subclass of ALL with poor prognosis. BCR-ABL1-expressing leukemic cells are highly dependent on double-strand break (DSB) repair signals for their survival. Here we report that a first-in-class HDAC1,2 selective inhibitor and doxorubicin (a hyper-CVAD chemotherapy regimen component) impair DSB repair networks in Ph+ B-cell precursor ALL cells using common as well as distinct mechanisms. The HDAC1,2 inhibitor but not doxorubicin alters nucleosomal occupancy to impact chromatin structure, as revealed by MNase-Seq. Quantitative mass spectrometry of the chromatin proteome along with functional assays showed that the HDAC1,2 inhibitor and doxorubicin either alone or in combination impair the central hub of DNA repair, the Mre11-Rad51-DNA ligase 1 axis, involved in BCR-ABL1-specific DSB repair signaling in Ph+ B-cell precursor ALL cells. HDAC1,2 inhibitor and doxorubicin interfere with DISC (DNA damage-induced transcriptional silencing in cis)) or transcriptional silencing program in cis around DSB sites via chromatin remodeler-dependent and -independent mechanisms, respectively, to further impair DSB repair. HDAC1,2 inhibitor either alone or when combined with doxorubicin decreases leukemia burden in vivo in refractory Ph+ B-cell precursor ALL patient-derived xenograft mouse models. Overall, our novel mechanistic and preclinical studies together demonstrate that HDAC1,2 selective inhibition can overcome DSB repair 'addiction' and provide an effective therapeutic option for Ph+ B-cell precursor ALL.

Activation of Ha-ras p21 by substitution, deletion, and insertion mutations.

The transforming activity of naturally arising ras oncogenes results from point mutations that affect residue 12 or 61 of the encoded 21-kilodalton protein (p21). By use of site-directed mutagenesis, we showed that deletions and insertions of amino acid residues in the region of residue 12 are also effective in conferring oncogenic activity on p21. Common to these various alterations is the disruption that they create in this domain of the protein, which we propose results in the inactivation of a normal function of the protein.

Naming in young children: a dumb attentional mechanism?

Previous studies have shown that young children selectively attend to some object properties and ignore others when generalizing a newly learned object name. Moreover, the specific properties children attend to depend on the stimulus and task context. The present study tested an attentional account: that children's feature selection in name generalization is guided by non-strategic attentional processes that are minimally influenced by new conceptual information presented in the task. Four experiments presented 3-year-old children and adults with novel artifacts consisting of distinctive base objects with appended parts. In a Name condition, subjects were asked whether test objects had the same name as the exemplar. In a Similarity condition, subjects made similarity judgments for the same objects. Subjects in two experiments were shown a function for either the base object or the parts. Both adults' naming and similarity judgments were influenced by the functional information. Children's similarity judgments were also influenced by the functions. However, children's naming was immune to influence from information about function. Instead, children's feature selection in naming was shifted only by changes in the relative salience of base objects and parts. The results are consistent with the idea that dumb attentional processes are responsible for young children's smart generalizations of novel words to new instances. Potential mechanisms to explain these findings are discussed.

Effects of laser in situ keratomileusis (LASIK) on the corneal endothelium.

PURPOSE: To assess the effects of laser in situ keratomileusis (LASIK) on the corneal endothelium. METHODS: In a prospective study, the corneal endothelium of 98 eyes of 65 consecutive patients (mean age, 41 years; range, 22 to 66 years) was photographed before, 2 weeks after, and 12 weeks after LASIK for the correction of 2.75 to 14.5 diopters of myopia. Theoretical ablation depths were 200 to 330 microm below the corneal surface. Cell density, coefficient of variation, and percent of hexagonal cells were determined using 150 to 200 cells from each image. Eighty-eight eyes (91%) of 59 patients had a history of contact lens wear. RESULTS: The mean+/-SD preoperative endothelial cell density was 2,549+/-365 cells per mm2, and the mean coefficient of variation was 0.35+/-0.06. There was no statistically significant change in the mean endothelial cell density or mean coefficient of variation of cell size at the 2-week (2,561+/-360 cells per mm2 and 0.35+/-0.06) or 12-week (2,541+/-364 cells per mm2 and 0.35+/-0.05) postoperative examinations. The percent of hexagonal cells was not significantly changed 2 weeks postoperatively; however, 12 weeks postoperatively (P=.0413, two-tailed t test), the percent of hexagonal cells was decreased by 1%. CONCLUSIONS: Corneal endothelial cell density and morphology were unchanged 2 and 12 weeks after LASIK for the correction of up to 14.5 diopters of myopia. In this LASIK study, the correction of up to 14.5 diopters of myopia appears to cause no clinically significant effect on corneal endothelial cell density or morphology.

A New Source of Resistance to Tapesia yallundae Associated with a Homoeologous Group 4 Chromosome in Thinopyrum ponticum.

ABSTRACT Wheat (Thinopyrum ponticum line SS767; PI 611939) with 42 chromosomes previously was identified as a new source of eyespot resistance. Individual plants of SS767 were tested for reaction to Tapesia yallundae, the major pathogen of eyespot in the Pacific Northwest region of the United States. Resistance of this line was similar to the resistant winter wheat cv. Madsen (carrying gene Pch1 for eyespot resistance). Polymerase chain reaction analysis with primers specific for the J or E genomes revealed that SS767 contains Thinopyrum chromatin. Cytological and Cbanding analyses demonstrated that SS767 is a chromosome substitution line in which wheat chromosome 4D is replaced by a homoeologous group 4 chromosome of Thinopyrum ponticum. Genomic in situ hybridization using St genomic DNA from Pseudoroegneria strigosa as a probe, which can differentiate chromosomes from different genomes of Thinopyrum, indicated that this chromosome belongs to the J genome. Molecular analysis of an F(2) population segregating for chromosome 4J and resistance to eyespot confirmed that eyespot resistance in line SS767 is associated with chromosome 4J of Thinopyrum ponticum. This is the first report of genetic control of resistance to eyespot derived from Thinopyrum ponticum. This source of resistance provides a new opportunity to improve wheat resistance to eyespot by adding to the diversity of resistance sources available.

Evaluation of Dasypyrum villosum Populations for Resistance to Cereal Eyespot and Stripe Rust Pathogens.

Resistance to Pseudocercosporella herpotrichoides (cause of eyespot) and Puccinia striiformis(cause of stripe rust) was evaluated in a germ plasm collection of Dasypyrum villosum (syn. Haynaldia villosa) and a set of disomic addition lines, a substitution, and a translocation line of D. villosum chromosomes in a wheat background. Three races of P. striiformis and a ß-glucuronidase-transformed strain of Pseudocercosporella herpotrichoides were used to inoculate plants and evaluate disease reactions. Of the 115 D. villosum accessions tested, 33 (28.6%) were resistant to one or more races of Puccinia striiformis and 8 accessions were resistant to all races. All 219 accessions of D. villosum tested were resistant to Pseudocercosporella herpotrichoides and 158 (72%) of the accessions had lower ß-glucuronidase activity than the resistant wheat line VPM-1. Most of the accessions of D. villosum resistant to the stripe rust pathogen originated from Greece; however, there was no distinction among origins for resistance to the eyespot pathogen. Chromosome 4V was confirmed to carry the gene for resistance to P. herpotrichoides. At least one gene for resistance to Puccinia striiformis was located on the short arm of chromosome 6V of D. villosum in the 6VS/6AL-translocation line; this gene was named Yr26.

Perennial Wheat Germ Plasm Lines Resistant to Eyespot, Cephalosporium Stripe, and Wheat Streak Mosaic.

A perennial wheat cropping system on the Palouse Prairie of eastern Washington may provide an alternative to the Federal Conservation Reserve Program and reduce soil erosion while providing a harvestable crop for growers. Twenty-four perennial wheat germ plasm lines resulting from crosses between wheat and wheatgrass were evaluated under controlled environment conditions for resistance to Wheat streak mosaic virus (WSMV), Cephalosporium gramineum, and Tapesia yallundae (anamorph Pseudocercosporella herpotrichoides var. herpotrichoides). Perennial wheat lines SS452, SS103, SS237, MT-2, and PI 550713 were resistant to all three pathogens. Eight lines (33%) were resistant to WSMV at 21°C and 25°C; AT3425 was resistant to WSMV at 21°C but not at 25°C. Thirteen lines (54%) were highly to moderately resistant to C. gramineum. Thirteen lines (54%) were resistant to T. yallundae in each experiment, but the reactions of four lines differed between experiments. The wheatgrasses Thinopyrum intermedium (PI 264770) and Thinopyrum ponticum (PI 206624) are reported as new sources of resistance to T. yallundae. Perennial wheat must have resistance to these diseases in order to be feasible as a crop in the Pacific Northwest.

Effects of exposure to aggressive behavior on job satisfaction of health care staff.

This study examined the effects of exposure to the aggressive and disruptive behavior of institutionalized elderly patients on health care staff's job satisfaction. Aggressive behavior was defined as physical, verbal, or general disruptive behavior. Health care staff from a geriatric long-term care hospital were interviewed about their exposure to these types of aggressive behavior and the effect of this exposure on job satisfaction. Job satisfaction was significantly correlated with overall exposure to aggressive behavior. Although physical aggression was reported more frequently, exposure to verbal aggression correlated higher with job satisfaction. Exposure to aggressive behavior was the best predictor of job satisfaction, followed by level of education. These findings indicate that job satisfaction is negatively affected by exposure to aggressive behavior but positively affected by educational level. The role of educational programs in moderating the impact of exposure to aggressive behavior on job satisfaction is discussed.

Cellulitis incidence in a defined population.

A population-based insurance claims database was used to examine cellulitis incidence, anatomical sites of infection, complicating diagnoses, source of health service, and recurrence rates. Insurance claim files were searched for cellulitis ICD-9-CM codes 681.0-682.9. Complications of cellulitis including erysipelas, lymphadenitis, lymphangitis, and necrotizing fasciitis were also identified by ICD-9-CM codes. We found a cellulitis incidence rate of 24.6/1000 person-years, with a higher incidence among males and individuals aged 45-64 years. The most common site of infection was the lower extremity (39.9%). The majority of patients were seen in an outpatient setting (73.8%), and most (82.0%) had only one episode of cellulitis during the 5-year period studied. There was a very low incidence of cellulitis complications, including necrotizing fasciitis. Cellulitis is fairly common, usually treated in outpatient settings, and is infrequently complicated by erysipelas, lymphadenitis, lymphangitis, or necrotizing fasciitis.

Resistance to eyespot of wheat, caused by Tapesia yallundae, derived from Thinopyrum intermedium homoeologous group 4 chromosome.

Thinopyrum intermedium was identified previously as resistant to Tapesia yallundae, cause of eyespot of wheat. Using GUS-transformed isolates of T. yallundae as inoculum, we determined that wheat lines carrying Th. intermedium chromosome 4 Ai#2 or the short arm of chromosome 4 Ai#2 were as resistant to the pathogen as the eyespot-resistant wheat- Th. ponticum chromosome substitution line SS 767 (PI 611939) and winter wheat cultivar Madsen, which carries gene Pch 1 for eyespot resistance. Chromosome 4 E from Th. elongatum and chromosome 4 J from Th. bessarabicum did not confer resistance to T. yallundae. Genome-specific PCR primers confirmed the presence of Thinopyrum chromatin in these wheat- Thinopyrum lines. Genomic in situ hybridization using an St genomic probe from Pseudoroegneria strigosa demonstrated that chromosome 4 Ai#2 belongs to the J(s) genome of Thinopyrum. The eyespot resistance in the wheat- Th. intermedium lines is thus controlled by the short arm of this J(s) chromosome. This is the first report of resistance to T. yallundae controlled by a J(s) genome chromosome of Th. intermedium.