Protein extraction from grape tissues by two-dimensional electrophoresis

At the onset of proteomic studies protein samples have to be accurately separated by two dimensional electrophoresis (2-DE); subsequently polypeptides are identified. Grape tissues, in particular roots, can be very problematic due to their hardness and to the high content of compounds that interfere in classical protein extraction. We have used a phenol-based extraction method in the presence of a protease inhibitor and Polyvinylpolypyrrolidone (PVPP). In this paper we demonstrate that this extraction method gives satisfactory and reproducible protein separation allowing the identification of some proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

Protein extraction from grape tussues by two-dimensional electrophoresis

At the onset of proteomic studies protein samples have to be accurately separated by two dimensional electrophoresis (2-DE); subsequently polypeptides are identified. Grape tissues, in particular roots, can be very problematic due to their hardness and to the high content of compounds that interfere in classical protein extraction. We have used a phenol-based extraction method in the presence of a protease inhibitor and Polyvinylpolypyrrolidone (PVPP). In this paper we demonstrate that this extraction method gives satisfactory and reproducible protein separation allowing the identification of some proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)

The polo-like kinase 1 (PLK1) inhibitor NMS-P937 is effective in a new model of disseminated primary CD56+ acute monoblastic leukaemia

CD56 is expressed in 15–20% of acute myeloid leukaemias (AML) and is associated with extramedullary diffusion, multidrug resistance and poor prognosis. We describe the establishment and characterisation of a novel disseminated model of AML (AML-NS8), generated by injection into mice of leukaemic blasts freshly isolated from a patient with an aggressive CD56+ monoblastic AML (M5a). The model reproduced typical manifestations of this leukaemia, including presence of extramedullary masses and central nervous system involvement, and the original phenotype, karyotype and genotype of leukaemic cells were retained in vivo. Recently Polo-Like Kinase 1 (PLK1) has emerged as a new candidate drug target in AML. We therefore tested our PLK1 inhibitor NMS-P937 in this model either in the engraftment or in the established disease settings. Both schedules showed good efficacy compared to standard therapies, with a significant increase in median survival time (MST) expecially in the established disease setting (MST = 28, 36, 62 days for vehicle, cytarabine and NMS-P937, respectively). Importantly, we could also demonstrate that NMS-P937 induced specific biomarker modulation in extramedullary tissues. This new in vivo model of CD56+ AML that recapitulates the human tumour lends support for the therapeutic use of PLK1 inhibitors in AML

Pig domestication and human-mediated dispersal in western eurasia revealed through ancient DNA and geometric morphometrics

Zooarcheological evidence suggests that pigs were domesticated in Southwest Asia ∼8,500 BC. They then spread across the Middle and Near East and westward into Europe alongside early agriculturalists. European pigs were either domesticated independently or more likely appeared so as a result of admixture between introduced pigs and European wild boar. As a result, European wild boar mtDNA lineages replaced Near Eastern/Anatolian mtDNA signatures in Europe and subsequently replaced indigenous domestic pig lineages in Anatolia. The specific details of these processes, however, remain unknown. To address questions related to early pig domestication, dispersal, and turnover in the Near East, we analyzed ancient mitochondrial DNA and dental geometric morphometric variation in 393 ancient pig specimens representing 48 archeological sites (from the Pre-Pottery Neolithic to the Medieval period) from Armenia, Cyprus, Georgia, Iran, Syria, and Turkey. Our results reveal the first genetic signatures of early domestic pigs in the Near Eastern Neolithic core zone. We also demonstrate that these early pigs differed genetically from those in western Anatolia that were introduced to Europe during the Neolithic expansion. In addition, we present a significantly more refined chronology for the introduction of European domestic pigs into Asia Minor that took place during the Bronze Age, at least 900 years earlier than previously detected. By the 5th century AD, European signatures completely replaced the endemic lineages possibly coinciding with the widespread demographic and societal changes that occurred during the Anatolian Bronze and Iron Ages

Unravelling the complexity of domestication:A case study using morphometrics and ancient DNA analyses of archaeological pigs from Romania

Funding statement. This work was supported by the Natural Environment Research Council (NE/F003382/1) and the Leverhulme Trust (F/00 128/AX) Acknowledgements. Archaeozoological analyses conducted by A. Ba˘la˘s¸escu were supported by three grants from the Romanian National Authority for Scientific Research, CNCS UEFISCDI (PN-II-RU-TE-20113-0146, PN-II-ID-PCE-2011-3-0982 and PN-IIID-PCE-2011-3-1015). We thank the archeologists Ca˘ta˘lin Bem, Alexandru Dragoman, Valentin Dumitras¸cu, Laura Dietrich, Raluca Koga˘lniceanu, Cristian Micu, Sta˘nica Pandrea, Valentin Parnic, George Trohani, Valentina Voinea for the material they generously provided. We thank the many institutions and individuals that provided sample material and access to collections, especially the curators of the Museum fu¨r Naturkunde, Berlin; Muse´um National d’Histoire Naturelle, Paris; Muse´um d’Histoire Naturelle, Gene`ve; Museum fu¨r Haustierkunde, Halle; National Museum of Natural History, Washington; The Field Museum, Chicago and The American Museum of Natural History, New York; The Naturhistorisches Museum, BernPeer reviewedPublisher PD

Imports and isotopes: a modern baseline study for interpreting Iron Age and Roman trade in fallow deer antlers

The European Fallow deer (Dama dama dama) became extinct in the British Isles and most of continental Europe at the time of the Last Glacial Maximum, with the species becoming restricted to an Anatolian refugium (Masseti et al. 2008). Human-mediated reintroductions resulted in fallow populations in Rhodes, Sicily, Mallorca, Iberia and other parts of western Europe (Sykes et al. 2013). Eventually, the species was brought to Britain by the Romans during the 1st century AD, with a breeding population being established at Fishbourne Roman Palace (Sykes et al. 2011). The human influence on the present-day distribution of the species makes it particularly interesting from a zooarchaeological perspective. This paper describes my MSc research, as part of the AHRC-funded project Dama International: Fallow Deer and European Society 6000 BC–AD 1600, looking at antlers from Iron Age and Roman sites in Britain for evidence of trade in body parts and whether this can be elucidated by a parallel stable isotope study of modern fallow antlers of known provenance

Effects of Anacetrapib in Patients with Atherosclerotic Vascular Disease

BACKGROUND: Patients with atherosclerotic vascular disease remain at high risk for cardiovascular events despite effective statin-based treatment of low-density lipoprotein (LDL) cholesterol levels. The inhibition of cholesteryl ester transfer protein (CETP) by anacetrapib reduces LDL cholesterol levels and increases high-density lipoprotein (HDL) cholesterol levels. However, trials of other CETP inhibitors have shown neutral or adverse effects on cardiovascular outcomes. METHODS: We conducted a randomized, double-blind, placebo-controlled trial involving 30,449 adults with atherosclerotic vascular disease who were receiving intensive atorvastatin therapy and who had a mean LDL cholesterol level of 61 mg per deciliter (1.58 mmol per liter), a mean non-HDL cholesterol level of 92 mg per deciliter (2.38 mmol per liter), and a mean HDL cholesterol level of 40 mg per deciliter (1.03 mmol per liter). The patients were assigned to receive either 100 mg of anacetrapib once daily (15,225 patients) or matching placebo (15,224 patients). The primary outcome was the first major coronary event, a composite of coronary death, myocardial infarction, or coronary revascularization. RESULTS: During the median follow-up period of 4.1 years, the primary outcome occurred in significantly fewer patients in the anacetrapib group than in the placebo group (1640 of 15,225 patients [10.8%] vs. 1803 of 15,224 patients [11.8%]; rate ratio, 0.91; 95% confidence interval, 0.85 to 0.97; P=0.004). The relative difference in risk was similar across multiple prespecified subgroups. At the trial midpoint, the mean level of HDL cholesterol was higher by 43 mg per deciliter (1.12 mmol per liter) in the anacetrapib group than in the placebo group (a relative difference of 104%), and the mean level of non-HDL cholesterol was lower by 17 mg per deciliter (0.44 mmol per liter), a relative difference of -18%. There were no significant between-group differences in the risk of death, cancer, or other serious adverse events. CONCLUSIONS: Among patients with atherosclerotic vascular disease who were receiving intensive statin therapy, the use of anacetrapib resulted in a lower incidence of major coronary events than the use of placebo. (Funded by Merck and others; Current Controlled Trials number, ISRCTN48678192 ; ClinicalTrials.gov number, NCT01252953 ; and EudraCT number, 2010-023467-18 .)

High-resolution luminescence speleothem records from Savi Cave, Trieste, NE Italy.

6High-resolution luminescence speleothem records from Savi Cave, Trieste, NE Italy - We measured three luminescent records from the sample SV1, a stalagmite from Savi Cave (Trieste karst, Italy). The longest panoramic record (having the lowest resolution from the records) is a proxy of the solar infl uence on the climatic system and covers last 14430±176 years. The step of the record varies from 1.1 to 12.7 years. First composite record, which consists of 81000 data points, is compiled of 39 overlapping scans. It covers the last 5005±140 years. The time step of the record varies from 9.9 days to 33.9 days. The highest resolution record covers last 2028±100 years and allows precise determination of growth rate of the stalagmite. It consists of 40106 data points compiled of 16 overlapping scans. Its time step varies from 15.6 days to 19.9 days. We made a reconstruction of the annual growth rate variations for the last 2028 years, which represents annual precipitation for this region.Record di luminescenza ad alta risoluzione di uno speleotema della Grotta Savi, Trieste, Italia - Si presentano tre record di luminescenza della stalagmite SV1 proveniente dalla Grotta Savi nel Carso di Trieste. Il record panoramico più lungo (e con la risoluzione più bassa) è un dato proxy dell’infl uenza solare sul clima locale, e copre gli ultimi 14430±176 anni, con una risoluzione che varia da 1,1 a 12,7 anni. Il primo record composito, costituito da 81000 dati assemblati da 39 scansioni sovrapposte, copre gli ultimi 5005±140 anni con una risoluzione che varia da 9,9 a 33,9 giorni. Il record a maggior risoluzione, costituito da 40106 dati assemblati da 16 scansioni sovrapposte, copre gli ultimi 2028±100 anni con una risoluzione che varia da 15,6 a 19,9 giorni, e consente di determinare con precisione il tasso di crescita annuale della stalagmite. In base a questo record si è ricostruito il tasso di crescita annuale della stalagmite che rappresenta il valore di precipitazione media annuale per l’area in esame.nonemixedSTOYKOVA D.; SHOPOV Y.; SAURO U.; BORSATO A.; CUCCHI F.; FORTI P.Stoykova, D.; Shopov, Y.; Sauro, U.; Borsato, A.; Cucchi, Franco; Forti, P