This trend has meant that relatively pristine landscapes are at increasingly greater risk from offsite contamination from the billions of tonnes of mine waste produced (Mudd, 2013). Evaluating recent mining influences on previously non-mining impacted systems enables greater insight into the short-term effects from environmental contamination compared to networks subjected to long-term cumulative damage (Hildén and Rapport, 1993 and Arkoosh et al., 1998). Given that river systems are the primary conduit for metal transport in catchments, their adjoining

environments are ideal for assessing upstream mining impacts and risks associated with their use. Metal mining pollutants that become stored in alluvial RGFP966 mw sediments can produce long-term risks to the environment (Miller, 1997, Hudson-Edwards et al., 2001, Macklin et al., 2003 and von der Heyden and New, 2004). These pollutants also provide potential pathways for exposure via the food chain (Miller et al., 2004). Therefore, evaluating and quantifying risks associated with off site mine waste provides guidance to users of environments that are subject to contamination (e.g. graziers, fisherman, irrigators, potable water extractors, cf. Foulds et al., 2014). Analysis of impact can also assist with the implementation of tighter regulatory regimes where necessary. The increase in environmental Ceritinib regulations

governing contemporary mining operations (as opposed to historic mining) suggests that the release of mine-contaminants into relatively pristine areas will likely be associated with instantaneous accidental spills, particularly during times of flood. In fact, during the past 40 years, 75 major spills of mining contaminated materials have released contaminated waters and sediments to river systems, averaging nearly two per year, mafosfamide not including those in secluded regions (Miller and Orbock Miller, 2007). Few studies have documented the downstream extent to which the contaminants affect ecosystem health, the trends in contaminant distributions that result

from these spills (Miller and Orbock Miller, 2007), or the potential short-and long-term environmental impacts that result. Even fewer spills have been studied along rivers devoid of previous mining activity generating contrasting results. Graf (1990), for example, found that the downstream transport and deposition of contaminated sediment resulting from the 1979 Church Rock uranium tailings spill led to a non-systematic downstream trend in 230Th concentrations. Rather, concentrations varied as a function of stream power and the duration over which shear stress exceeded critical values along the channel. In contrast, the 1998 Aznalcóllar Mine spill in Spain generated a high sediment-laden flow that produced a semi-systematic downstream decrease in the thickness of the deposited, mine-contaminated sediment (Gallart et al., 1999).

Strong archeological evidence suggests that the islands within the northern

Lagoon have been inhabited since Roman times and up to the Medieval Age. Examples of wooden waterside structures were found dating back between the first century BC and the second century AD (Canal, 1998, Canal, 2013 and Fozzati, 2013). As explained in Housley et al. (2004), due to the need for dry land suitable for building, salt marshes were enclosed and infilled to support small islands on which early settlements were built. Sites that go back to Roman imperial times are now well documented in the northern part of the lagoon. In the city of Venice itself, however, the first archeological evidence found www.selleckchem.com/products/ldk378.html so far dates back to the 5th century AD. Only later, in the 8th to 9th century AD, did Venice start to take the character of a city (Ammerman, 2003). By the end of the 13th century, Venice was a prosperous city with a population of about 100,000 inhabitants (Housley et al., 2004). At the beginning of the 12th century, sediment delivered by the system of rivers threatened to fill the lagoon (Gatto and Carbognin, 1981). In the short term, the infilling of sediment affected the navigation and harbor activity of Venice, while in the long term,

it opened up the city to military attack by land. This situation motivated the Venetians to divert the rivers away from the lagoon, so that the sediment load of the rivers would discharge directly into the Protein tyrosine phosphatase Adriatic Sea. This human intervention was carried out over the next few centuries so that all the main rivers Tenofovir flowing into the lagoon were diverted by the 19th century (Favero, 1985 and Bondesan and Furlanetto, 2012). If the Venetians had not

intervened, the fate of the Venice Lagoon could have been the same as that of a lagoon in the central part of the Gulf of Lions in the south of France. This lagoon was completely filled between the 12th and 13th century (Sabatier et al., 2010). In the 19th century, significant modifications included a reduction of the number of inlets from eight to three. The depth of the remaining inlets also increased from ∼5 m to ∼15 m, with a consequent increase in tidal flow and erosive processes (Gatto and Carbognin, 1981). In the last century, dredging of major navigation channels took place in the central part of the lagoon to enhance the harbor activity. The exploitation of underground water for the industrial area of Marghera (Fig. 1) contributed to a sinking of the bottom of the basin (Carbognin, 1992 and Brambati et al., 2003). Also, the lagoon surface decreased by more than 30 percent due to activities associated with land reclamation and fish-breeding. The morphological and ecological properties of the lagoon changed dramatically: salt marsh areas decreased by more than 50 percent (from 68 km2 in 1927 to 32 km2 in 2002) and some parts of the lagoon deepened (Carniello et al., 2009, Molinaroli et al., 2009 and Sarretta et al.

Specific types of cancer may not grow as efficiently in mouse bone as they do in a human microenvironment, hence the need for humanized models [90]. This general approach is reflected into varied attempts to explore the homing of prostate cancer (manuscript in preparation), myeloma cells [91], leukemia [92], and breast cancer cells [93] to humanized microenvironments. Stem cells in bone bring forth a remarkable change of perspective in bone medicine. They allow for consideration of diseases that affect bone as an organ rather than as a tissue. They provide

the tool needed to understand diseases of the skeleton other GDC-0199 concentration than osteoporosis, while also contributing to the understanding of osteoporosis and bone aging. They provide a novel angle, centered on bone progenitors, in the study of major hematological diseases. They open the prospect of understanding the interaction of bone and cancer using the understanding of the HME/niche as a blueprint. Finally,

pursuing these avenues of strict medical relevance can advance our understanding of bone disease, which can feed back on our understanding of bone physiology. Personal work mentioned in this article was supported by Telethon Foundation (GGP09227), MIUR (20102M7T8X), Fondazione Roma (2008), Fondazione Cenci Bolognetti (103/2011), Ministry of Health of Italy (G21J11000040001), EU (PluriMes consortium602423) and Sapienza University of Rome (C26A11LF98; C26A12TKEZ), and by the DIR, NIDCR, of the IRP, NIH, DHHS (PGR) (1ZIADE000380). “
“The publisher regrets that Fig. 2 was published incorrectly. The correct figure appears PFT�� BCKDHB below. The publisher would like to apologise for any inconvenience caused. Figure options Download full-size image Download high-quality image (609 K) Download as PowerPoint slide “
“The authors regret that the acknowledgements were published incorrectly. They should read as follows: Funding statement: This study was funded by Merck & Co., Inc., Whitehouse Station, NJ, USA. Acknowledgment: We thank Gregg Wesolowski, Parvithra Ramakrishnan (Merck & Co.) and Aurora Varela and

Susan Smith (Charles River Laboratories) for their technical assistance for this study. We would also like to thank the LAR staff (Merck & Co.) for providing care for the animals in this study. We would also like to thank Tara Cusick and Boyd B Scott (Merck & Co.) for their critical review of this manuscript. Finally we would like to thanks Jennifer Pawlowski (Merck & Co.) for the logistical support during the submission of this manuscript. COI statement: Author RS is an employee of Charles Rivers Laboratories which conducted contract research for Merck & Co for this study. Authors DYS and HD received consultant fees from Merck & Co. Authors MAG and LTD are employees of Merck & Co. and may own stock in the company. Author CH was an employee of Merck & Co at the time the study was conducted.

First-Dimension Isoelectric Focusing was conducted using the Ettan IPGphor Cup Loading Manifold (GE Healthcare) and the following voltage PLX3397 molecular weight settings: 150 V constant 2 h, 300 V constant 3 h, ramp to 600 V 3 h, ramp to 2000 V 3 h, ramp 8000 V 3 h, constant 8000 V 3 h 20 min, to reach a total of 48 kV h. Strips were stored at −80 °C until further processing.

Prior to the second dimension SDS-PAGE, IPG strips were equilibrated for 15 min in urea/SDS equilibration/reduction buffer (6 M urea, 30% glycerol (w/v), 2% SDS (w/v), 50 mM Tris/HCL (pH 8.8), 0.007% bromophenol blue (BFB) and 65 mM DTT) and followed by 15 min of alkylation in a similar buffer containing 259 mM iodoacetamide instead of DTT. The equilibrated IPG strips were rinsed in Tris-Glycine/SDS running buffer (Bio-Rad) and positioned onto 10–15% gradient acrylamide gels (Sigma–Aldrich Optigel

no bind silane, A116230) and then sealed by 0.5% (w/v) agarose overlay solution. Gels were run in a Dodeca Cell running tank (Bio-Rad) filled with Tris-Glycine/SDS running buffer. Temperature was set to 24 °C and proteins were allowed to separate selleck screening library at a constant current of 10 mA/gel for 1 h in the dark, followed by 60 mA/gel until the 10 kDa band of the Kaleidoscope marker (Bio-Rad, 161-0375) had reached the bottom of the gels. Cy2, Cy3 and Cy5 images were acquired from each gel using a Typhoon scanner 9400 (GE Healthcare) with the following PMT voltage settings: Cy2, 435 V; Cy3, 435 V; and Cy5, 400 V. Gel image files were analyzed using Progenesis SameSpots software version 3.1 (Non Linear Dynamics) with default settings. Match vectors were automatically generated and subsequently checked manually and complemented. A total of 1804 individual protein spots were detected, quantified and matched Farnesyltransferase through all gel images. Over 1500 of these spots showed coefficient of variation (CV) for the quantitative values below 10% in 4 technical replicates

(labeling and running two Cy3 and two Cy5 internal standard samples). Preparative 2D-gels with up to 340 μg of unlabeled myotube protein (mixed samples from T2D and NGT subjects) was run and stained with SYPRO Ruby (Invitrogen) and spots were visualized using a laser scanner (FX Pro, Bio-Rad). The protein profile from previous analytical 2-D DIGE gels (CyDye-labeled samples) and the preparative gels were carefully matched with PDQuest image analysis software (Bio-Rad). Protein spots found to contain differential protein abundance in myotubes derived from T2D versus NGT subjects were excised and pooled from three preparative 2D-PAGE gels using the ExQuest robot equipped with a 1.5 mm punch tool (Bio-Rad). Gel plug pieces were destained (70% ACN, 25 mM NH4HCO3) and dried. Proteins were digested overnight at 37 °C with trypsin in 25 mM NH4HCO3 (Promega). Trypsin fragments were analyzed using an LC/MS system consisting of a 1200 Series liquid chromatograph, HPLC-Chip Cube MS interface and a 6510 QTOF mass spectrometer (Agilent Technologies).

Os corticoides e os anti-histamínicos podem ser usados no tratamento das formas ligeiras e moderadas. A necessidade de os inibidores da protease serem ingeridos com alimentos faz com que a disgueusia seja um sintoma que deve ser monitorizado com cuidado. O boceprevir e, sobretudo, o telaprevir são metabolizados pelo citocromo p450 3A4 e 3A5 (CYP3A4/5). Existe, portanto, o risco de interação com medicamentos metabolizados pelas mesmas vias. Assim, chamamos a atenção para fármacos que podem induzir o CYP3A e, desse modo, baixar a concentração plasmática dos inibidores da protease como, por exemplo, a rifampicina, fenitoína e a Proteasome assay carbamazepina; outros medicamentos são inibidores, competitivos ou não,

do CYP3A, diminuindo o metabolismo do boceprevir e do telaprevir, originando a sua sobredosagem como, por

exemplo, os antifúngicos. Os inibidores da protease podem, por outro lado, ter um efeito inibidor sobre diversos medicamentos, o que pode originar sobredosagem desses fármacos, como buy MG-132 é o caso dos antiarrítmicos amiodarona, os derivados da ergotamina, as benzodiazepinas e as estatinas; outros, ainda, podem ter a sua eliminação aumentada com a consequente redução da sua eficácia, como é o caso dos anovulatórios, escitalopram, desipramina e zolpidem. Consoante as situações, durante a terapêutica com os inibidores da protease pode ser necessário descontinuar alguns fármacos ou proceder a modificações da dosagem. “
“Diarreia crónica define-se como uma alteração no trânsito intestinal caracterizada pela alteração da consistência das fezes, aumento do número de frequência das dejeções (mais de dejeções diárias) e peso fecal superior a 200 g/24 h, prologando-se por mais de

4 semanas. O diagnóstico diferencial pode ser muito complexo e abrangente, pois pode ter inúmeras etiologias: causas infecciosas, PFKL endócrino-metabólicas, neoplásicas, funcionais e medicamentosas. Doente do sexo feminino, de 46 anos, caucasiana, residente em Leiria. Referenciada à consulta de Medicina Interna por diarreia crónica com estudo inconclusivo, episódios de lipotimia e incontinência esfincteriana. Referia o início do quadro há 8 anos (1997) com cerca de 6 dejeções diarreicas/dia, líquidas, por vezes com resíduos alimentares, diurnas e noturnas, sem sangue, sem muco e sem tenesmo ou falsas vontades. Negava fatores desencadeantes ou outros sintomas acompanhantes, nomeadamente náuseas, vómitos, febre, artralgias, alterações cutâneas, dor abdominal, esteatorreia. Os antecedentes pessoais eram irrelevantes. Do historial familiar, a doente era filha única de pai incógnito e negava patologia relevante da linha materna. Não existia igualmente registo de viagens recentes ou hábitos medicamentosos previamente ao início da diarreia. Dos exames complementares, realizados na altura, fez hemograma, bioquímica completa, estudo hormonal incluindo FSH, LH, PTH, cortisol, TSH, T3 e T4 totais e livres, que se encontravam dentro dos valores normais.

While it is still unclear how flexibly these distributed cortical networks contribute to semantic processing across different task contexts (Mahon and Caramazza, 2008, Pulvermueller, 2013 and Willems and Casasanto, 2011), evidence suggests convincingly that sensorimotor activation in response to printed words reflects semantic processing. In UK primary schools, children learn to read simple words during their first year when they

are 4–5 years old. Reading fluency continues to develop substantially after that, with improvements in reading speed and accuracy extending until around the 15th year of life (Wechlser, 2001). Age-related changes in reading skills are accompanied by focalisation and left-lateralisation of word shape selective occipito-temporal areas (Brown et al., 2005, Schlaggar and McCandliss, 2007 and Schlaggar et al., 2002) and decreasing activation in posterior temporal areas associated with cross-modal orthographic and check details phonological processing (Church et al., 2008 and Pugh et al., 2001).

While substantial research has charted how structural and functional changes in these language-related areas contribute to reading improvement during development, the role of cortical sensorimotor learn more representations in this process has not yet been explored. It is therefore unclear when printed words start engaging the same brain areas as their pictorial counterparts as children learn to decode meaning from word forms. Understanding this process can provide important insight into how and under which circumstances child readers access the sensorimotor meaning of written words, and provide a benchmark for investigating word comprehension in children

with reading difficulties. This research can also inform theories on how distributed semantic sensorimotor networks contribute to the printed word-learning process. Only a few studies have investigated distributed semantic networks in the developing sensorimotor cortex, but initial evidence suggests that these might already be present before Mirabegron children learn to read. For instance, by 6–7 years of age, passive viewing of tool pictures without the overt plan to act, engages grasp-related areas of the cortex whilst passive viewing of animal pictures does not (Dekker, Mareschal, Sereno, & Johnson, 2011). Similarly, by 4 years of age, listening to actions words (verbs) activates motor areas in the brain, but listening to non-action words (nouns) does not (James & Maouene, 2009). Which role might such already-established cortical sensorimotor representations play during reading acquisition? It is possible that sensorimotor networks become involved early during reading training, for example because they may help bootstrap the formation of mappings between word shape and word meaning (Nation, 2008). Or, underdeveloped spelling/sound connections might allow for a greater influence of semantic information on slow word-recognition processes (Plaut & Booth, 2000).

Two different acceptor beads can be used: with AlphaScreen acceptor beads, final emission is from rubrene (λem=520–620 nm); with AlphaLISA acceptor beads, the final emission is from europium that is doped into the beads which shows a much narrower emission spectrum (λem=615 nm). For protein kinases,

typically a biotinylated peptide substrate is conjugated to streptavidin coated donor beads and a phosphospecific antibody is conjugated to protein A coated acceptor bead ( Von Leoprechting et al., 2004). Because long wavelength light is used for excitation and emission occurs at shorter wavelengths, optical interference of the excitation light by compounds

is reduced. However, AlphaScreen has been shown to be susceptible to compound interference by color quenchers of the emission light as well as anti-oxidants, singlet oxygen selleck compound quenchers, find protocol and biotin mimetics if streptavidin coated beads are used ( Baell and Holloway, 2010). Another consideration in developing AlphaScreen assays is that variation of the biotinylated peptide substrate will show a “hook-effect” (as mentioned above) at high substrate concentrations due to saturation of the streptavidin donor bead binding sites – the signal first increases with increasing peptide, then levels off and starts to decrease when excess peptide is used as the proportion of productive donor acceptor bead complexes decreases due to the excess peptide in the assay ( Quinn et al., 2010). Therefore, it is not possible to determine Km values for the peptide substrate using AlphaScreen as binding capacity of the beads limits the detectable range of substrate concentration. An advantage of this detection strategy is that the large distance dependence (~200 nm) allows the employment much of physiologically relevant substrates. Analytical approaches to kinase detection include microfluidic

systems for separation based on electrophoretic and electro-osmotic properties of the labeled species (Cohen et al., 1999). In one such technology platform (Caliper, PerkinElmer), optimization of the elution gradient on the UPLC system allows one to run kinase assays at extremely low substrate conversion rates (Wu et al., 2006). In this system, separation and quantification of substrate and product is provided and detection and interfering fluorescent compounds are readily flagged. With generic systems available that detect ATP/ADP conversion one could ask if there is any advantage to protein kinase assay systems that rely on detection of phosphorylated peptides, especially when peptide-based systems are oftentimes incapable of incorporating natural protein substrates.

Coverage was assessed as a percentage of the sea bottom covered by vegetation or a certain species within the extent of the sampling site.

Along the transects, the total coverage of the macrovegetation community, coverage of individual species and character of substrate were registered visually by the diver or recorded with an underwater video camera. Observations were carried out to the deepest limit of vegetation on the transect. In the Kõiguste and buy GSK458 Sõmeri areas, 8–10 observations were made along the transects (the deepest vegetation at 10 m depth). In the Orajõe area the number of observations per transect was 7–9 (the deepest vegetation at 8.3 m depth). Paired with the sampling of seabed phytobenthic community in May, July and September, beach wrack samples were also collected in April, June, August and October (Table 1). Wrack samples were collected from three transects parallel to the shoreline in each area. The distance between the transects was about 60 m. The selleck chemicals lengths of the transects were 5 m and five samples were collected from each transect. The samples were collected using a 20 cm × 20 cm metal frame at

a distance of 1 m from one another. Each individual frame sample served as a sampling unit in further statistical analyses. This design (3 transects and 5 samples per transect) resulted in 15 samples per area in each month. Distances from the water edge [m], thickness [cm] and coverage [%] of the wrack layer inside the sampling frame were measured.

The freshest beach wrack closest to the Beta adrenergic receptor kinase sea was always chosen for sampling. As a rule, older, more or less decomposed wrack strips were located higher on the shore. In April, only three samples were collectable from fresh beach cast material. As the rest of the samples included old material cast ashore during the previous autumn before the sea froze up, the April data were excluded from further quantitative analyses. The collected material was packed and kept frozen. In the laboratory, the species composition in each sample was determined. As wrack specimens were often fragmented and detailed identification was impossible, morphologically very similar species were treated as one group. The filamentous brown algae Ectocarpus siliculosus (Dillwyn) Lyngbye and Pilayella littoralis (Linnaeus) Kjellman were not separated. All characeans except Tolypella nidifica (O.F. Müller) Leonhardi were determined as Chara spp. Higher plants with similar morphology such as Zannichellia palustris L., Ruppia maritima L. and Stuckenia pectinata (L.) Börner were treated as one group. The biomasses of Fucus vesiculosus L. and Furcellaria lumbricalis (Hudson) J. V. Lamouroux and the rest of the sample were separated and weighed after drying at 60°C to constant weight. Biomass (grams dry weight) was calculated per square metre [g d.w. m−2]).

The optimal concentration of HRP-conjugated streptavidin was determined in the same way. The calibrator consisted of the culture supernatant TSA HDAC mouse from DG44 CHO cells expressing recombinant CL-11. A two-fold serial dilution of the culture supernatant was used to generate an eight-point calibrator curve with a range from 0.26 to 34.8 ng/ml. A five-parameter fit model was applied to the calibrating samples and used to estimate the concentration of unknown samples. The calibrator was stored as single-use aliquots at − 80 °C. The QCs consisted of a pool of serum or plasma from five healthy volunteers diluted 1/11, 1/80 and 1/500 in dilution buffer to

represent high, medium and low concentrations of CL-11, respectively. The QCs were stored as single ready-to-use aliquots at − 80 °C. To study parallelism, the calibrator serial dilution curve was compared to the serial dilution curves of two batches of purified recombinant CL-11 and serial dilutions curves of plasma and serum from two blood donors (analyzed in duplicates). OD data were GDC-0980 evaluated using regression analysis on logistically transformed values, an algorithm that comprised several steps. Due to the maximum limit of the OD determination,

a number of consecutive measurements of OD = 4.0 was observed in each dilution series. Only the last value of OD = 4.0 was maintained in each dilution series, while the prior maximum determinations were omitted.

Subsequently, all OD values were divided by 4.1 to transform the OD data to values above 0, but below 1, as required for the subsequent logistic transformation, y′ = ln[y/(1 − y)]. A background level of OD = 0.05 was observed, and values below the corresponding logistically transformed values were omitted from further analysis. A linear regression was fitted to the remaining data points and multiple comparisons among slopes using Tukey’s HSD test were used to compare the parallelism of the different serial dilutions. The statistical analyses were performed using the Analyse-it software (Analyse-it Software, Ltd, Leeds, UK). Ten two-fold serial dilutions of serum and plasma samples from five blood donors were analyzed in triplicates. Coefficients of variation Y-27632 2HCl (CV) were calculated for the triplicate measurements of each dilution. A “measured/mean” ratio was expressed for each sample using the triplicate measurements and calculating the mean of the triplicates. To study linearity, the CL-11 concentration calculated for each dilution and multiplied by the dilution factor was compared to a mean of the CL-11 concentration that was back-calculated from four dilutions of each sample (1/16–1/128 for serum samples and 1/20–1/160 for plasma samples). The working range was determined as the CL-11 concentrations for which CV was < 10% and the measured/mean ratios deviated < 20%.

15 In some virus infections, causal therapy is not possible; in others, such as CMV infection in immunocompetent individuals, antiviral therapy is not necessary because the CAS is slight and self-remitting. The efficacy of corticosteroid therapy has not been systematically investigated, and opinions promoted in textbooks and review articles differ widely.[3], [14], [15] and [69] We treated a 45 year old man admitted with Mycoplasma pneumonia and an initial Hgb level of 3.5 g/dL due to an anti-I mediated severe CAS. He received erythromycin and high-dose prednisolone

followed by rapid tapering of the corticosteroid dose. His condition improved rapidly and he became transfusion independent within days. Similar case Pexidartinib supplier reports have been published in the literature. [100] and [101] Since spontaneous resolution eventually occurs in all patients, however, guidelines can hardly be built upon case

observations. Therefore, there is no documentation for using corticosteroids routinely in CAS secondary to infection. Until more data are provided, corticosteroid therapy may be considered if the hemolysis is severe and spontaneous improvement does not occur selleck kinase inhibitor within some days. Plasmapheresis may be helpful in selected, extreme cases. [72] and [102] If indicated, erythrocyte transfusions can safely be given provided the same precautions are undertaken as in primary CAD. 31 Diagnosing the subtype Staurosporine mw of AIHA precisely is essential for the choice of therapy. The molecular, immunological and immunohistochemical characteristics of the clonal lymphoproliferation in primary CAD should be further studied. The authors declare no financial or other conflicts of interest. “
“Polycythemia vera (PV) and essential thrombocythemia (ET) are chronic myeloproliferative neoplasms (MPN) characterized by clonal expansion of an abnormal hematopoietic stem/progenitor cell. Natural history of these MPN is marked by thrombo-hemorrhagic complications and a propensity to transform into myelofibrosis and acute leukemia. Understanding

of the pathophysiology of these disorders dramatically improved following the description in 2005 of recurrent molecular abnormalities represented by: the V617F mutation in JAK2 exon 14, that is the most frequent and involves > 95% of PV and about 60–70% of ET patients [1], [2], [3] and [4]; a number of molecular alterations located in JAK2 exon 12 [5], [6] and [7]; mutations in MPL, mostly represented by the W515L or W515K allele, which are present in about 7% of ET patients. [8] and [9] These and other new genetic notions have modified our criteria for diagnosis, prognosis and therapy although it is still unclear whether these concepts can be translated into changes of the management of individual patients with MPN.