Late Maastrichtian carbon isotope stratigraphy and cyclostratigraphy of the Newfoundland Margin (Site U1403, IODP Expedition 342)

Earth’s climate during the Maastrichtian (latest Cretaceous) was punctuated by brief warming and cooling episodes, accompanied by perturbations of the global carbon cycle. Superimposed on a long-term cooling trend, the middle Maastrichtian is characterized by deep-sea warming and relatively high values of stable carbon-isotope ratios, followed by strong climatic variability towards the end of the Cretaceous. A lack of knowledge on the timing of climatic change inhibits our understanding of underlying causal mechanisms. We present an integrated stratigraphy from Integrated Ocean Drilling Program (IODP) Site U1403, providing an expanded deep ocean record from the North Atlantic (Expedition 342, Newfoundland Margin). Distinct sedimentary cyclicity suggests that orbital forcing played a major role in depositional processes, which is confirmed by statistical analyses of high resolution elemental data obtained by X-ray fluorescence (XRF) core scanning. Astronomical calibration reveals that the investigated interval encompasses seven 405-kyr cycles (Ma4051 to Ma4057) and spans the 2.8 Myr directly preceding the Cretaceous/Paleocene (K/Pg) boundary. A high-resolution carbon-isotope record from bulk carbonates allows us to identify global trends in the late Maastrichtian carbon cycle. Low-amplitude variations (up to 0.4‰) in carbon isotopes at Site U1403 match similar scale variability in records from Tethyan and Pacific open-ocean sites. Comparison between Site U1403 and the hemipelagic restricted basin of the Zumaia section (northern Spain), with its own well-established independent cyclostratigraphic framework, is more complex. Whereas the pre-K/Pg oscillations and the negative values of the Mid-Maastrichtian Event (MME) can be readily discerned in both the Zumaia and U1403 records, patterns diverge during a ~ 1 Myr period in the late Maastrichtian (67.8–66.8 Ma), with Site U1403 more reliably reflecting global carbon cycling. Our new carbon isotope record and cyclostratigraphy offer promise for Site U1403 to serve as a future reference section for high-resolution studies of late Maastrichtian paleoclimatic change

Recent outbreaks of infectious syphilis, United Kingdom, January 2012 to April 2014

Six outbreaks of infectious syphilis in the United Kingdom, ongoing since 2012, have been investigated among men who have sex with men (MSM) and heterosexual men and women aged under 25 years. Interventions included case finding and raising awareness among healthcare professionals and the public. Targeting at-risk populations was complicated as many sexual encounters involved anonymous partners. Outbreaks among MSM were influenced by the use of geospatial real-time networking applications that allow users to locate other MSM within close proximity

Hsp27 regulates podocyte cytoskeletal changes in an in vitro model of podocyte process retraction

Nephrotic syndrome (NS) is characterized by structural changes in the actin‐rich foot processes of glomerular podocytes. We previously identified high concentrations of the small heat shock protein hsp27 within podocytes as well as increased glomerular accumulation and phosphorylation of hsp27 in puromycin aminonucleoside (PAN) ‐induced experimental NS. Here we analyzed murine podocytes stably transfected with hsp27 sense, antisense, and vector control constructs using a newly developed in vitro PAN model system. Cell morphology and the microfilament structure of untreated sense and antisense transfectants were altered compared with controls. Vector cell survival, polymerized actin content, cell area, and hsp27 content increased after 1.25 μg/ml PAN treatment and decreased after 5.0 μg/ml treatment. In contrast, sense cells were unaffected by 1.25 μg/ml PAN treatment whereas antisense cells showed decreases or no changes in all parameters. Treatment of sense cells with 5.0 μ g/ml PAN resulted in increased cell survival and cell area whereas antisense cells underwent significant decreases in all parameters. Hsp27 provided dramatic protection against PAN‐induced microfilament disruption in sense > vector > antisense cells. We conclude that hsp27 is able to regulate both the morphological and actin cytoskeletal response of podocytes in an in vitro model of podocyte injury.—Smoyer, W. E., Ransom, R. F. Hsp27 regulates podocyte cytoskeletal changes in an in vitro model of podocyte process retraction. FASEB J. 16, 315–326 (2002)Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/154256/1/fsb2fj010681com.pd

Validation of a HLA-A2 tetramer flow cytometric method, IFNgamma real time RT-PCR, and IFNgamma ELISPOT for detection of immunologic response to gp100 and MelanA/MART-1 in melanoma patients

<p>Abstract</p> <p>Background</p> <p>HLA-A2 tetramer flow cytometry, IFNγ real time RT-PCR and IFNγ ELISPOT assays are commonly used as surrogate immunological endpoints for cancer immunotherapy. While these are often used as research assays to assess patient's immunologic response, assay validation is necessary to ensure reliable and reproducible results and enable more accurate data interpretation. Here we describe a rigorous validation approach for each of these assays prior to their use for clinical sample analysis.</p> <p>Methods</p> <p>Standard operating procedures for each assay were established. HLA-A2 (A*0201) tetramer assay specific for gp100_209(210M)and MART-1_26–35(27L), IFNγ real time RT-PCR and ELISPOT methods were validated using tumor infiltrating lymphocyte cell lines (TIL) isolated from HLA-A2 melanoma patients. TIL cells, specific for gp100 (TIL 1520) or MART-1 (TIL 1143 and TIL1235), were used alone or spiked into cryopreserved HLA-A2 PBMC from healthy subjects. TIL/PBMC were stimulated with peptides (gp100₂₀₉, gp100_pool, MART-1_27–35, or influenza-M1 and negative control peptide HIV) to further assess assay performance characteristics for real time RT-PCR and ELISPOT methods. Validation parameters included specificity, accuracy, precision, linearity of dilution, limit of detection (LOD) and limit of quantification (LOQ). In addition, distribution was established in normal HLA-A2 PBMC samples. Reference ranges for assay controls were established.</p> <p>Results</p> <p>The validation process demonstrated that the HLA-A2 tetramer, IFNγ real time RT-PCR, and IFNγ ELISPOT were highly specific for each antigen, with minimal cross-reactivity between gp100 and MelanA/MART-1. The assays were sensitive; detection could be achieved at as few as 1/4545–1/6667 cells by tetramer analysis, 1/50,000 cells by real time RT-PCR, and 1/10,000–1/20,000 by ELISPOT. The assays met criteria for precision with %CV < 20% (except ELISPOT using high PBMC numbers with %CV < 25%) although flow cytometric assays and cell based functional assays are known to have high assay variability. Most importantly, assays were demonstrated to be effective for their intended use. A positive IFNγ response (by RT-PCR and ELISPOT) to gp100 was demonstrated in PBMC from 3 melanoma patients. Another patient showed a positive MART-1 response measured by all 3 validated methods.</p> <p>Conclusion</p> <p>Our results demonstrated the tetramer flow cytometry assay, IFNγ real-time RT-PCR, and INFγ ELISPOT met validation criteria. Validation approaches provide a guide for others in the field to validate these and other similar assays for assessment of patient T cell response. These methods can be applied not only to cancer vaccines but to other therapeutic proteins as part of immunogenicity and safety analyses.</p

How to Adapt to Changing Markets: Experience and Personality in a Repeated Investment Game

Investment behavior is traditionally investigated with the assumption that it is on average advantageous to invest. However, this may not always be the case. In this paper, we experimentally studied investment choices made by students and financial professionals facing alternately an advantageous and disadvantageous environment in a multi-round investment game. Expected returns from investment in the advantageous environment were higher than a safe alternative, while expected returns were lower in the disadvantageous environment. We investigate how experience and personality are related to choices. Investment behavior does not differ dependent on expected returns and professionals do not significantly differ from students. Personality predicts behavior and in particular we observe that openness to experience was an asset in unfavorable markets, leading to reduced risk taking

Validation of a HLA-A2 tetramer flow cytometric method, IFNgamma real time RT-PCR, and IFNgamma ELISPOT for detection of immunologic response to gp100 and MelanA/MART-1 in melanoma patients

<p>Abstract</p> <p>Background</p> <p>HLA-A2 tetramer flow cytometry, IFNγ real time RT-PCR and IFNγ ELISPOT assays are commonly used as surrogate immunological endpoints for cancer immunotherapy. While these are often used as research assays to assess patient's immunologic response, assay validation is necessary to ensure reliable and reproducible results and enable more accurate data interpretation. Here we describe a rigorous validation approach for each of these assays prior to their use for clinical sample analysis.</p> <p>Methods</p> <p>Standard operating procedures for each assay were established. HLA-A2 (A*0201) tetramer assay specific for gp100_209(210M)and MART-1_26–35(27L), IFNγ real time RT-PCR and ELISPOT methods were validated using tumor infiltrating lymphocyte cell lines (TIL) isolated from HLA-A2 melanoma patients. TIL cells, specific for gp100 (TIL 1520) or MART-1 (TIL 1143 and TIL1235), were used alone or spiked into cryopreserved HLA-A2 PBMC from healthy subjects. TIL/PBMC were stimulated with peptides (gp100₂₀₉, gp100_pool, MART-1_27–35, or influenza-M1 and negative control peptide HIV) to further assess assay performance characteristics for real time RT-PCR and ELISPOT methods. Validation parameters included specificity, accuracy, precision, linearity of dilution, limit of detection (LOD) and limit of quantification (LOQ). In addition, distribution was established in normal HLA-A2 PBMC samples. Reference ranges for assay controls were established.</p> <p>Results</p> <p>The validation process demonstrated that the HLA-A2 tetramer, IFNγ real time RT-PCR, and IFNγ ELISPOT were highly specific for each antigen, with minimal cross-reactivity between gp100 and MelanA/MART-1. The assays were sensitive; detection could be achieved at as few as 1/4545–1/6667 cells by tetramer analysis, 1/50,000 cells by real time RT-PCR, and 1/10,000–1/20,000 by ELISPOT. The assays met criteria for precision with %CV < 20% (except ELISPOT using high PBMC numbers with %CV < 25%) although flow cytometric assays and cell based functional assays are known to have high assay variability. Most importantly, assays were demonstrated to be effective for their intended use. A positive IFNγ response (by RT-PCR and ELISPOT) to gp100 was demonstrated in PBMC from 3 melanoma patients. Another patient showed a positive MART-1 response measured by all 3 validated methods.</p> <p>Conclusion</p> <p>Our results demonstrated the tetramer flow cytometry assay, IFNγ real-time RT-PCR, and INFγ ELISPOT met validation criteria. Validation approaches provide a guide for others in the field to validate these and other similar assays for assessment of patient T cell response. These methods can be applied not only to cancer vaccines but to other therapeutic proteins as part of immunogenicity and safety analyses.</p

Characterization of Mycosphaerellaceae species associated with citrus greasy spot in Panama and Spain

[EN] Greasy spot of citrus, caused by Zasmidium citri-griseum (= Mycosphaerella citri), is widely distributed in the Caribbean Basin, inducing leaf spots, premature defoliation, and yield loss. Greasy spot-like symptoms were frequently observed in humid citrus-growing regions in Panama as well as in semi-arid areas in Spain, but disease aetiology was unknown. Citrus-growing areas in Panama and Spain were surveyed and isolates of Mycosphaerellaceae were obtained from citrus greasy spot lesions. A selection of isolates from Panama (n = 22) and Spain (n = 16) was assembled based on their geographical origin, citrus species, and affected tissue. The isolates were characterized based on multi-locus DNA (ITS and EF-1 alpha) sequence analyses, morphology, growth at different temperatures, and independent pathogenicity tests on the citrus species most affected in each country. Reference isolates and sequences were also included in the analysis. Isolates from Panama were identified as Z. citri-griseum complex, and others from Spain attributed to Amycosphaerella africana. Isolates of the Z. citri-griseum complex had a significantly higher optimal growth temperature (26.8 degrees C) than those of A. africana (19.3 degrees C), which corresponded well with their actual biogeographical range. The isolates of the Z. citri-griseum complex from Panama induced typical greasy spot symptoms in 'Valencia' sweet orange plants and the inoculated fungi were reisolated. No symptoms were observed in plants of the 'Ortanique' tangor inoculated with A. africana. These results demonstrate the presence of citrus greasy spot, caused by Z. citri-griseum complex, in Panama whereas A. africana was associated with greasy spot-like symptoms in Spain.Research was partially funded by 'Programa de Formacion de los INIA Iberoamerica' and INIA RTA2010-00105-00-00-FEDER to Vidal Aguilera Cogley.. We thank J. Martinez-Minaya (UV) for assistance with INLAAguilera-Cogley, VA.; Berbegal Martinez, M.; Català, S.; Collison Brentu, F.; Armengol Fortí, J.; Vicent Civera, A. (2017). Characterization of Mycosphaerellaceae species associated with citrus greasy spot in Panama and Spain. PLoS ONE. 12(12):1-19. https://doi.org/10.1371/journal.pone.0189585S1191212Crous, P. W., Summerell, B. A., Carnegie, A. J., Wingfield, M. J., Hunter, G. C., Burgess, T. I., … Groenewald, J. Z. (2009). Unravelling Mycosphaerella: do you believe in genera? Persoonia - Molecular Phylogeny and Evolution of Fungi, 23(1), 99-118. doi:10.3767/003158509x479487Mondal, S. N., & Timmer, L. W. (2006). Greasy Spot, a Serious Endemic Problem for Citrus Production in the Caribbean Basin. Plant Disease, 90(5), 532-538. doi:10.1094/pd-90-0532Whiteside, J. O. (1970). Etiology and Epidemiology of Citrus Greasy Spot. Phytopathology, 60(10), 1409. doi:10.1094/phyto-60-1409Huang, F., Groenewald, J. Z., Zhu, L., Crous, P. W., & Li, H. (2015). Cercosporoid diseases of Citrus. Mycologia, 107(6), 1151-1171. doi:10.3852/15-059Wellings, C. R. (1981). Pathogenicity of fungi associated with citrus greasy spot in New South Wales. Transactions of the British Mycological Society, 76(3), 495-499. doi:10.1016/s0007-1536(81)80080-0Marco, G. M. (1986). A Disease Similar to Greasy Spot but of Unknown Etiology on Citrus Leaves in Argentina. Plant Disease, 70(11), 1074a. doi:10.1094/pd-70-1074aVidal Aguilera-Cogley, & Antonio Vicent. (2015). FUNGAL DISEASES OF CITRUS IN PANAMA. Acta Horticulturae, (1065), 947-952. doi:10.17660/actahortic.2015.1065.118Honger J. Aetiology and importance of foliage diseases affecting citrus in the nursery at the Agricultural Research Station (ARS). PhD Thesis. Accra: University of Ghana; 2004.Vicent A, Álvarez A, León M, García-Jiménez J. Mycosphaerella sp. asociada a manchas foliares de cítricos en España. In: Proceedings of the 13th Congress of the Spanish Phytopathological Society. 2006; Murcia; Spain.Abdelfattah, A., Cacciola, S. O., Mosca, S., Zappia, R., & Schena, L. (2016). Analysis of the Fungal Diversity in Citrus Leaves with Greasy Spot Disease Symptoms. Microbial Ecology, 73(3), 739-749. doi:10.1007/s00248-016-0874-xQuaedvlieg, W., Binder, M., Groenewald, J. Z., Summerell, B. A., Carnegie, A. J., Burgess, T. I., & Crous, P. W. (2014). Introducing the Consolidated Species Concept to resolve species in the Teratosphaeriaceae. Persoonia - Molecular Phylogeny and Evolution of Fungi, 33(1), 1-40. doi:10.3767/003158514x681981Edgar, R. C. (2004). MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research, 32(5), 1792-1797. doi:10.1093/nar/gkh340Darriba, D., Taboada, G. L., Doallo, R., & Posada, D. (2012). jModelTest 2: more models, new heuristics and parallel computing. Nature Methods, 9(8), 772-772. doi:10.1038/nmeth.2109Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D. L., Darling, A., Höhna, S., … Huelsenbeck, J. P. (2012). MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice Across a Large Model Space. Systematic Biology, 61(3), 539-542. doi:10.1093/sysbio/sys029Rambaut A. FigTree v1. 4.0, a graphical viewer of phylogenetic trees. Edinburgh, Scotland: University of Edinburgh; 2016.Spiegelhalter, D. J., Best, N. G., Carlin, B. P., & van der Linde, A. (2002). Bayesian measures of model complexity and fit. Journal of the Royal Statistical Society: Series B (Statistical Methodology), 64(4), 583-639. doi:10.1111/1467-9868.00353Rue, H., Martino, S., & Chopin, N. (2009). Approximate Bayesian inference for latent Gaussian models by using integrated nested Laplace approximations. Journal of the Royal Statistical Society: Series B (Statistical Methodology), 71(2), 319-392. doi:10.1111/j.1467-9868.2008.00700.xChristensen RH. Ordinal—regression models for ordinal data. R package version 2015.1–21. 2015. http://www.cran.r-project.org/package=ordinal/ Accessed 8 May 2017.Hunter, G. C., Wingfield, B. D., Crous, P. W., & Wingfield, M. J. (2006). A multi-gene phylogeny for species of Mycosphaerella occurring on Eucalyptus leaves. Studies in Mycology, 55, 147-161. doi:10.3114/sim.55.1.147Braun, U., & Urtiaga, R. (2013). New species and new records of cercosporoid hyphomycetes from Cuba and Venezuela (Part 2). Mycosphere, 4(2), 172-214. doi:10.5943/mycosphere/4/2/3Braun, U., Crous, P. W., & Nakashima, C. (2014). Cercosporoid fungi (Mycosphaerellaceae) 2. Species on monocots (Acoraceae to Xyridaceae, excluding Poaceae). IMA Fungus, 5(2), 203-390. doi:10.5598/imafungus.2014.05.02.04Aptroot A. Mycosphaerella and its anamorphs: conspectus of Mycosphaerella CBS Biodiversity Series 5. Utrecht: CBS-KNAW Fungal Biodiversity Centre; 2006.Crous, P. W., & Wingfield, M. J. (1996). Species of Mycosphaerella and Their Anamorphs Associated with Leaf Blotch Disease of Eucalyptus in South Africa. Mycologia, 88(3), 441. doi:10.2307/3760885Aguín, O., Sainz, M. J., Ares, A., Otero, L., & Pedro Mansilla, J. (2013). Incidence, severity and causal fungal species of Mycosphaerella and Teratosphaeria diseases in Eucalyptus stands in Galicia (NW Spain). Forest Ecology and Management, 302, 379-389. doi:10.1016/j.foreco.2013.03.021Maxwell, A., Dell, B., Neumeister-Kemp, H. G., & Hardy, G. E. S. J. (2003). Mycosphaerella species associated with Eucalyptus in south-western Australia: new species, new records and a key. Mycological Research, 107(3), 351-359. doi:10.1017/s0953756203007354Otero L, Aguín O, Mansilla J, Hunter G, Wingfield M. Identificación de especies de Mycosphaerella en Eucalyptus globulus y E. nitens en Galicia. In: Proceedings of the 13th Congress of the Spanish Phytopathological Society; 2006; Murcia, Spain.ZHAN, J., & McDONALD, B. A. (2011). Thermal adaptation in the fungal pathogen Mycosphaerella graminicola. Molecular Ecology, 20(8), 1689-1701. doi:10.1111/j.1365-294x.2011.05023.xPeel, M. C., Finlayson, B. L., & McMahon, T. A. (2007). Updated world map of the Köppen-Geiger climate classification. Hydrology and Earth System Sciences, 11(5), 1633-1644. doi:10.5194/hess-11-1633-200