Supplementary Table 2. Characteristics of the AML patients according to FLT3 and NPM1 gene mutational status.

Supplementary Figure 2. MPFC detection limit.

Supplementary Figure 3. MPFC and molecular analysis of UPN 241 bone marrow samples at diagnosis, during follow-up and at relapse.

Supplementary Table 5. Longitudinal analysis of FLT3-ITDs using patient-specific RQPCR in four AML patients.

Supplementary Figure 1. ROC curve analysis using CD34CD123CD25+ve immunophenotype.

Supplementary Table 1. anti-human antibodies used.

Adenosine A2A receptors are highly expressed in caudate-putamen, and external globus pallidus (GPe), but in trace in internal globus pallidus/entopeduncular nucleus. A2A upregulation in striatal-pallidal pathway is associated with levodopa dyskinesias in Parkinson's disease. Instead, A2A downregulation has been reported in striatum of Huntington Disease. We investigated distribution of A2A receptors in basal ganglia of Tor1a+/− knock-out mouse model DYT1 primary dystonia.

Familial dysautonomia (FD) is an inherited neurological disorder characterized by deficits in the peripheral sensory and autonomic systems. Almost all cases of FD occur in Jews of Ashkenazic extraction. The pattern of inheritance of FD is autosomal recessive and the disease tends to occur in siblings rather than in successive generations. A disturbance in catecholamine metabolism in patients with FD was first documented by the finding of high homovanillic acid and low vanillylmandelic acid urinary excretion rates indicating catecholamine insufficiency with shunting of the precursor to homovanillic acid. The finding of common autonomic symptoms in FD and in Parkinson's disease has suggested a possible neurochemical link between these two diseases. Although the involvement of the catecholaminergic system has been well documented in the pathogenesis of FD, further investigations are needed to clarify the molecular mechanism by which a genetic alteration produces the variety of autonomic signs observed in FD.

Two articles published 5 years ago concluded that the genome of the lizard Anolis carolinensis is an amniote genome without isochores. This claim was apparently contradicting previous results on the general presence of an isochore organization in all vertebrate genomes tested (including Anolis). In this investigation, we demonstrate that the Anolis genome is indeed heterogeneous in base composition, since its macrochromosomes comprise isochores mainly from the L2 and H1 families (a moderately GC-poor and a moderately GC-rich family, respectively), and since the majority of the sequenced microchromosomes consists of H1 isochores. These families are associated with different features of genome structure, including gene density and compositional correlations (e.g., GC3 vs flanking sequence GC and intron GC), as in the case of mammalian and avian genomes. Moreover, the assembled Anolis chromosomes have an enormous number of gaps, which could be due to sequencing problems in GC-rich regions of the genome. In conclusion, the Anolis genome is no exception to the general rule of an isochore organization in the genomes of vertebrates (and other eukaryotes).

Poly (ADP-ribose) polymerase 1 (PARP-1) is a nuclear enzyme that is involved in physiological processes as DNA repair, genomic stability, and apoptosis. Moreover, published studies demonstrated that PARP-1 mediates necrotic cell death in response to excessive DNA damage under certain pathological conditions. In Huntington's disease brains, PARP immunoreactivity was described in neurons and in glial cells, thereby suggesting the involvement of apoptosis in HD. In this study, we sought to determine if the PARP-1 inhibitor exerts a neuroprotective effect in R6/2 mutant mice, which recapitulates, in many aspects, human HD. Transgenic mice were treated with the PARP-1 inhibitor INO-1001 mg/Kg daily starting from 4 weeks of age. After transcardial perfusion, histological and immunohistochemical studies were performed. We found that INO 1001-treated R6/2 mice survived longer and displayed less severe signs of neurological dysfunction than the vehicle treated ones. Primary outcome measures such as striatal atrophy, morphology of striatal neurons, neuronal intranuclear inclusions and microglial reaction confirmed a neuroprotective effect of the compound. INO-1001 was effective in significantly increasing activated CREB and BDNF in the striatal spiny neurons, which might account for the beneficial effects observed in this model. Our findings show that PARP-1 inhibition could be considered as a valid therapeutic approach for HD.

This tribute is a modest but deeply felt homage to Christiane Groeben, ''the historical memory'' of the Stazione Zoologica Anton Dohrn.The work she has done over 45 years is a fundamental contribution to the history of the Stazione and to the history of biology.The debt that we all owe to her is multilayered, as a developing organism.At the inner layer we find her long and hard work as an archivist at the Stazione, which has made available to scholars a huge amount of relevant archives (manuscripts, correspondence, administrative documentation, photos) of an invaluable utility for a great number of historians and philosophers of biology as well as for art historians and historians of scientific institutions.In the intermediate layer we find her methodological and stringent lesson on the use of archival sources: each document is a fact, a fundamental starting point for each historical reconstruction, but at the same time this fact always needs a theory to be explained and accurately placed within a rational reconstruction of the past.Finally, the external and most apparent layer is anchored to Christiane's own research and publications, that have contributed to a deep knowledge of one of the most fantastic scientific enterprises, the creation of the Zoological Station, and to a better understanding of the personality of its creator, Anton Dohrn.

The human genome is a mosaic of isochores, which are long (>200 kb) DNA sequences that are fairly homogeneous in base composition and can be assigned to five families comprising 33%–59% of GC composition. Although the compartmentalized organization of the mammalian genome has been investigated for more than 40 years, no satisfactory automatic procedure for segmenting the genome into isochores is available so far. We present a critical discussion of the currently available methods and a new approach called isoSegmenter which allows segmenting the genome into isochores in a fast and completely automatic manner. This approach relies on two types of experimentally defined parameters, the compositional boundaries of isochore families and an optimal window size of 100 kb. The approach represents an improvement over the existing methods, is ideally suited for investigating long-range features of sequenced and assembled genomes, and is publicly available at https://github.com/bunop/isoSegmenter .

How the same DNA sequences can function in the three-dimensional architecture of interphase nucleus, fold in the very compact structure of metaphase chromosomes and go precisely back to the original interphase architecture in the following cell cycle remains an unresolved question to this day. The strategy used to address this issue was to analyze the correlations between chromosome architecture and the compositional patterns of DNA sequences spanning a size range from a few hundreds to a few thousands Kilobases. This is a critical range that encompasses isochores, interphase chromatin domains and boundaries, and chromosomal bands. The solution rests on the following key points: 1) the transition from the looped domains and sub-domains of interphase chromatin to the 30-nm fiber loops of early prophase chromosomes goes through the unfolding into an extended chromatin structure (probably a 10-nm "beads-on-a-string" structure); 2) the architectural proteins of interphase chromatin, such as CTCF and cohesin sub-units, are retained in mitosis and are part of the discontinuous protein scaffold of mitotic chromosomes; 3) the conservation of the link between architectural proteins and their binding sites on DNA through the cell cycle explains the "mitotic memory" of interphase architecture and the reversibility of the interphase to mitosis process. The results presented here also lead to a general conclusion which concerns the existence of correlations between the isochore organization of the genome and the architecture of chromosomes from interphase to metaphase.

Purpose: We evaluated leukemia-associated immunophenotypes (LAIP) and their correlation with fms-like tyrosine kinase 3 (FLT3) and nucleophosmin (NPM1) gene mutational status in order to contribute a better identificationof patients at highest risk of relapse in acute myeloid leukemia (AML). Experimental Design: Bone marrow samples from 132 patients with AML were analyzed by nine-color multiparametric flow cytometry. We confirmed the presence of the mutation in diagnostic samples and in sorted cells by conventional RT-PCR and by patient-specific RQ-PCR. Results:Within theCD34þ cell fraction, we identified a discrete population expressing high levels of the IL3 receptor a-chain (CD123) and MIC-2 (CD99) in combination with the IL2 receptor a-chain (CD25). The presence of this population positively correlated with the internal tandem duplications (ITD) mutation in the FLT3 gene (r 1⁄4 0.71). Receiver operating characteristics showed that, within the CD34þ cell fraction a percentage of CD123/CD99/CD25þ cells 11.7% predicted FLT3–ITD mutations with a specificity and sensitivity of >90%. CD34/CD123/ CD99/CD25þ clones were also detectable at presentation in 3 patients with FLT3 wild-type/NPM1þ AML who relapsed with FLT3-ITD/NPM1þ AML. Quantitative real-time PCR designed at relapse for each FLT3-ITD in these three cases confirmed the presence of low copy numbers of the mutation in diagnostic samples. Conclusions: Our results suggest that the CD34/CD25/ CD123/CD99þ LAIP is strictly associated with FLT3-ITD–positive cells. Clin Cancer Res; 21(17); 3977–85. 2015 AACR.

Second messenger cAMP and cGMP represent a key step in the action of dopamine that modulates directly or indirectly their synthesis. We aimed to verify whether levodopa-induced dyskinesias are associated with changes of the time course of levodopa/dopamine stimulated cAMP and cGMP levels, and/or with changes of their catabolism by phosphodiesterase activity in rats with experimental hemiparkinsonism. Microdialysis and tissue homogenates of the striatal tissues demonstrated that extracellular and intracellular cAMP/cGMP levels were lower in dyskinetic animals during the increasing phase of dyskinesias compared to eukinetic animals, but cAMP/cGMP levels increased in dyskinetic animals during the phase of decreasing and extinction of dyskinesias. Dyskinesias and the abnormal lowering of striatal cGMP and cAMP after levodopa were prevented by pretreatment with the multipotent drug amantadine, outlining the inverse relationship of cAMP/cGMP to dyskinesias. Moreover, dyskinetic animals showed higher striatal hydrolyzing cGMP-phosphodiesterase but not hydrolyzing cAMP-phosphodiesterase activity, suggesting that low cGMP but not cAMP levels could be due to increased catabolism. However, expressions of isozyme phosphodiesterase-1B and -10A highly and specifically located in the basal ganglia were not changed after levodopa in dyskinetic and eukinetic animals: accordingly, selective inhibitors of phosphodiesterase-1B and -10A were ineffective on levodopa dyskinesias. Therefore, the isozyme(s) expressing higher cGMP-phosphodiesterase activity in the striatum of dyskinetic animal should be determined. These observations suggest that dopamine-mediated processes of synthesis and/or degradation of cAMP/cGMP could be acutely impaired in levodopa dyskinesias, opening new ways to understanding physiopathology and treatment.

CD8 ϩ T cells from patients with acute multiple sclerosis display selective increase of adhesiveness in brain venules: a critical role for P-selectin glycoprotein ligand-1 Multiple sclerosis (MS) is considered an autoimmune inflammatory disease of the central nervous system. Under physi-ologic conditions, we compared the adhe-siveness of CD4 ؉ and CD8 ؉ lymphocytes from nontreated patients with acute, relapsing remitting multiple sclerosis (RRMS) and from healthy donors. We show that in patients with RRMS CD8 ؉ , but not with RRMS CD4 ؉ , T cells display increased rolling and arrest in inflamed murine brain venules. Moreover, CD8 ؉ , but not CD4 ؉ , lymphocytes from MS patients show increased rolling on P-selec-tin in vitro. Anti–P-selectin glycoprotein ligand-1 (PSGL-1) antibodies dramatically block the recruitment of CD8 ؉ cells in brain vessels of patients with MS, suggesting that PSGL-1 represents a novel pharmaceutical target that may be exploited to block the selective entrance of CD8 ؉ cells during early inflammation. Vas-cular cell adhesion molecule-1 (VCAM-1), but not PSGL-1, is critical for the adhesion of CD4 ؉ cells in MS patients, highlighting a fundamental dichotomy in the mechanisms governing the recruitment of lymphocyte subsets in RRMS. Importantly , 7-color fluorescence-activated cell sorter (FACS) analysis, together with functional data, indicates that a large fraction of CD8 ؉ cells from MS patients display the characteristics of memory-effector phenotype. In conclusion, our results show that CD8 ؉ , but not CD4 ؉ , T cells from patients with RRMS in the acute phase of the disease display increased ability to be recruited in inflamed brain venules.

BACKGROUND:The genomes of multicellular eukaryotes are compartmentalized in mosaics of isochores, large and fairly homogeneous stretches of DNA that belong to a small number of families characterized by different average GC levels, by different gene concentration (that increase with GC), different chromatin structures, different replication timing in the cell cycle, and other different properties. A question raised by these basic results concerns how far back in evolution the compartmentalized organization of the eukaryotic genomes arose.RESULTS:In the present work we approached this problem by studying the compositional organization of the genomes from the unicellular eukaryotes for which full sequences are available, the sample used being representative. The average GC levels of the genomes from unicellular eukaryotes cover an extremely wide range (19%-60% GC) and the compositional patterns of individual genomes are extremely different but all genomes tested show a compositional compartmentalization.CONCLUSIONS:The average GC range of the genomes of unicellular eukaryotes is very broad (as broad as that of prokaryotes) and individual compositional patterns cover a very broad range from very narrow to very complex. Both features are not surprising for organisms that are very far from each other both in terms of phylogenetic distances and of environmental life conditions. Most importantly, all genomes tested, a representative sample of all supergroups of unicellular eukaryotes, are compositionally compartmentalized, a major difference with prokaryotes.