Biomedical subjects

Y Z Gu

Publications and source records attributed to Y Z Gu.

At least 19 recordsLinked to original sources

PubMed2000

The basic helix-loop-helix-PAS protein MOP9 is a brain-specific heterodimeric partner of circadian and hypoxia factors.

PAS (PER, ARNT, SIM) proteins play important roles in adaptation to low atmospheric and cellular oxygen levels, exposure to certain environmental pollutants, and diurnal oscillations in light and temperature. In an attempt to better understand how organisms sense environmental changes, we have characterized a novel member of the PAS superfamily, MOP9 (member of PAS superfamily), that maps to human chromosome 12p11.22-11.23. This protein displays significant homology to the Drosophila circadian factor CYCLE and its putative mammalian ortholog MOP3/bMAL1. Like its homologs, MOP9 forms a transcriptionally active heterodimer with the circadian CLOCK protein, the structurally related MOP4, and hypoxia-inducible factors, such as HIF1alpha. In a manner consistent with its role as a biologically relevant partner of these proteins, MOP9 is coexpressed in regions of the brain such as the thalamus, hypothalamus, and amygdala. Importantly, MOP9 is coexpressed with CLOCK in the suprachiasmatic nucleus, the site of the master circadian oscillator in mammals.

ARNTL Transcription Factors↗

PubMed2000

The PAS superfamily: sensors of environmental and developmental signals.

Over the past decade, PAS domains have been identified in dozens of signal transduction molecules and various forms have been found in animals, plants, and prokaryotes. In this review, we summarize this rapidly expanding research area by providing a detailed description of three signal transduction pathways that utilize PAS protein heterodimers to drive their transcriptional output. It is hoped that these model pathways can provide a framework for use in understanding the biology of the less well-understood members of this emerging superfamily, as well as of those to be characterized in the days to come. We use this review to develop the idea that most eukaryotic PAS proteins can be classified by functional similarities, as well as by predicted phylogenetic relationships. We focus on the alpha-class proteins, which often act as sensors of environmental signals, and the beta-class proteins, which typically act as broad-spectrum partners that target these heterodimers to their genomic targets.

Amino Acid Sequence↗

PubMed1999

Cross-talk between the aryl hydrocarbon receptor and hypoxia inducible factor signaling pathways. Demonstration of competition and compensation.

The aryl hydrocarbon receptor (AHR) and the alpha-class hypoxia inducible factors (HIF1alpha, HIF2alpha, and HIF3alpha) are basic helix-loop-helix PAS (bHLH-PAS) proteins that heterodimerize with ARNT. In response to 2,3,7,8-tetrachlorodibenzo-p-dioxin, the AHR. ARNT complex binds to "dioxin responsive enhancers" (DREs) and activates genes involved in the metabolism of xenobiotics, e.g. cytochrome P4501A1 (Cyp1a1). The HIF1alpha.ARNT complex binds to "hypoxia responsive enhancers" and activates the transcription of genes that regulate adaptation to low oxygen, e.g. erythropoietin (Epo). We postulated that activation of one pathway would inhibit the other due to competition for ARNT or other limiting cellular factors. Using pathway specific reporters in transient transfection assays, we observed that DRE driven transcription was markedly inhibited by hypoxia and that hypoxia responsive enhancer driven transcription was inhibited by AHR agonists. When we attempted to support this cross-talk model using endogenous loci, we observed that activation of the hypoxia pathway inhibited Cyp1a1 up-regulation, but that activation of the AHR actually enhanced the induction of Epo by hypoxia. To explain this unexpected additivity, we examined the Epo gene and found that its promoter harbors DREs immediately upstream of its transcriptional start site. These experiments outline conditions where inhibitory and additive cross-talk occur between the hypoxia and dioxin signal transduction pathways and identify Epo as an AHR-regulated gene.

Base Sequence↗

PubMed1998

The basic-helix-loop-helix-PAS orphan MOP3 forms transcriptionally active complexes with circadian and hypoxia factors.

We report that MOP3 is a general dimerization partner for a subset of the basic-helix-loop-helix (bHLH)-PER-ARNT-SIM (PAS) superfamily of transcriptional regulators. We demonstrated that MOP3 interacts with MOP4, CLOCK, hypoxia-inducible factor 1alpha (HIF1alpha), and HIF2alpha. A DNA selection protocol revealed that the MOP3-MOP4 heterodimer bound a CACGTGA-containing DNA element. Transient transfection experiments demonstrated that the MOP3-MOP4 and MOP3-CLOCK complexes bound this element in COS-1 cells and drove transcription from a linked luciferase reporter gene. We also deduced the high-affinity DNA binding sites for MOP3-HIF1alpha complex (TACGTGA) and used transient transfection experiments to demonstrate that the MOP3-HIF1alpha and MOP3-HIF2alpha heterodimers bound this element, drove transcription, and responded to cellular hypoxia. Finally, we found that MOP3 mRNA expression overlaps in a number of tissues with each of its four potential partner molecules in vivo.

Animals↗

PubMed1998

Molecular characterization and chromosomal localization of a third alpha-class hypoxia inducible factor subunit, HIF3alpha.

Hypoxia inducible factors (HIFs) are heterodimeric transcription factors that regulate a number of adaptive responses to low oxygen tension. They are composed of alpha- and beta-subunits that belong to the basic helix-loop-helix-PAS (bHLH-PAS) superfamily. In our efforts to identify new bHLH-PAS proteins, we cloned a cDNA encoding a novel alpha-class hypoxia inducible factor, HIF3alpha. The HIF3alpha open reading frame encodes a 662-amino acid protein with a predicted molecular weight of 73 kDa and is expressed in adult thymus, lung, brain, heart, and kidney. The N-terminal bHLH-PAS domain of this protein shares amino acid sequence identity with that of HIF1alpha and HIF2alpha (57% and 53% identity, respectively). The C-terminus of HIF3alpha contains a 36-amino acid sequence that shares 61% identity with the hypoxia responsive domain-1 (HRD1) of HIF1alpha. In transient transfections, this domain confers hypoxia responsiveness when linked to a heterologous transactivation domain. In vitro studies reveal that HIF3alpha dimerizes with a prototype beta-class subunit, ARNT, and that the resultant heterodimer recognizes the hypoxia responsive element (HRE) core sequence, TACGTG. Transient transfection experiments demonstrate that the HIF3alpha-ARNT interaction can occur in vivo, and that the activity of HIF3alpha is upregulated in response to cobalt chloride or low oxygen tension.

Amino Acid Sequence↗

PubMed1997

Characterization of a subset of the basic-helix-loop-helix-PAS superfamily that interacts with components of the dioxin signaling pathway.

In an effort to better understand the mechanism of toxicity of 2,3,7, 8-tetrachlorodibenzo-p-dioxin, we employed an iterative search of human expressed sequence tags to identify novel basic-helix-loop-helix-PAS (bHLH-PAS) proteins that interact with either the Ah receptor (AHR) or the Ah receptor nuclear translocator (ARNT). We characterized five new "members of the PAS superfamily," or MOPs 1-5, that are similar in size and structural organization to the AHR and ARNT. MOPs 1-4 have N-terminal bHLH and PAS domains and C-terminal variable regions. MOP5 contained the characteristic PAS domain and a variable C terminus; it is possible that the cDNA contains a bHLH domain, but the entire open reading frame has yet to be completed. Coimmunoprecipitation studies, yeast two-hybrid analysis, and transient transfection experiments demonstrated that MOP1 and MOP2 dimerize with ARNT and that these complexes are transcriptionally active at defined DNA enhancer sequences in vivo. MOP3 was found to associate with the AHR in vitro but not in vivo. This observation, coupled with the fact that MOP3 formed tighter associations with the 90-kDa heat shock protein than the human AHR, suggests that MOP3 may be a conditionally active bHLH-PAS protein that requires activation by an unknown ligand. The expression profiles of the AHR, MOP1, and MOP2 mRNAs, coupled with the observation that they all share ARNT as a common dimeric partner, suggests that the cellular pathways mediated by MOP1 and MOP2 may influence or respond to the dioxin signaling pathway.

Amino Acid Sequence↗

PubMed1997

S49 cells endogenously express subtype 2 somatostatin receptors which couple to increase protein tyrosine phosphatase activity in membranes and down-regulate Raf-1 activity in situ.

S49 cells expressed type 2 somatostatin receptors (sstr2) by immunoblotting. Analysis by reverse transcription and polymerase chain reaction (RT-PCR) methodologies showed that S49 cells express predominantly sstr2A and sstr2B mRNAs; other subtypes were either not detected, in the case of sstr1, sstr3, sstr4, or variably detected, in the case of sstr5. No mutations were present in S49 cells at codon 12, 13, or 61 of the N-, K-, or H-ras genes. Nevertheless, randomly growing S49 cells contained Raf-1 activity by specific immune complex kinase assays. Treatment of S49 cells with somatostatin transiently inactivated the basal activity of Raf-1, but not that of B-Raf. Addition of somatostatin plus guanyl-5'-yl imidodiphosphate (GMPPNP) to S49 membranes stimulated PTPase activity. The concentration dependence for stimulation of PTPase activity correlated with high affinity binding of [125I-Tyr11]somatostatin-14. Both the effect of somatostatin to stimulate PTPase activity and to inactivate Raf-1 were abrogated by PTx. PTPase activity stimulated by somatostatin plus GMPPNP was recovered in a peak of high apparent M(r) (670,000) after solubilisation with Triton X-100 and Superose 6 chromatography. Furthermore, addition of activated, brain G alpha i/o subunits to fractions from control membranes stimulated PTPase activity in the high M(r) peak. Thus, S49 membranes contain a G-protein regulated PTPase (PTPase-G), and PTPase-G in these cells may reside in a high molecular weight complex.

Animals↗

PubMed1997

Coupling specificity between somatostatin receptor sst2A and G proteins: isolation of the receptor-G protein complex with a receptor antibody.

Somatostatin initiates its actions via a family of seven-transmembrane domain receptors. Of the five somatostatin receptor genes cloned, sst2 exists as two splice variants with the sst2A isoform being predominantly expressed. This receptor is widely distributed in endocrine, exocrine, and neuronal cells, as well as in hormonally responsive tumors, and leads to inhibition of secretion, electrical excitability, and cell proliferation. To investigate the specificity of signal transduction by the sst2A receptor, we developed antibodies against two overlapping peptides located within the C terminus of the receptor protein: peptide 2C(SG), containing amino acids 334-348, and peptide 2C(ER), containing amino acids 339-359. Although antibodies to both peptides bound the inducing antigen with high affinity, only the antibodies against peptide 2C(ER) precipitated the receptor. The best antibody, R2-88, precipitated about 80% of the sst2A receptor-ligand complex solubilized from transfected CHO cells and was specific for the sst2A receptor isotype. Addition of GTPgammaS (10 microM) to the immunoprecipitated ligand-sst2A receptor complex markedly accelerated ligand dissociation, indicating that G proteins remained functionally associated with the receptor in the immunoprecipitate. Analysis of the G proteins coprecipitated with the sst2A receptor by immunoblotting with G protein antibodies showed that both G(alpha) and G(beta) subunits were bound to the hormone-receptor complex. Immunoprecipitation of the receptor was not affected by the presence of bound ligand. However, G protein subunits were coprecipitated only with the hormone-occupied receptor. Thus, the unoccupied receptor has low affinity for G proteins, and hormone binding stabilizes the receptor-G protein complex. Use of subtype-specific G protein antisera further showed that G alpha(i1), G alpha(i2), and G alpha(i3) were complexed with the sst2A receptor whereas Galpha(o), G alpha(z), and G alpha(q) were not. Together, these studies demonstrate that the sst2A receptor interacts selectively with G alpha(i) proteins in a hormone-dependent manner. The finding that this receptor couples to all three G alpha(i) subunits may help explain how somatostatin can regulate multiple signaling pathways.

Amino Acid Sequence↗

PubMed1997

Molecular characterization of the murine Hif-1 alpha locus.

Hypoxia inducible factor 1 alpha (HIF-1 alpha) is a basic helix-loop-helix-PAS (bHLH-PAS) transcription factor that mediates certain cellular responses to low oxygen tension, iron chelators, Co2+, Ni2+, Mg2+, and low intracellular glucose concentration. Upon exposure to the above conditions, HIF-1 alpha is upregulated and heterodimerizes with the Ah receptor nuclear translocator (ARNT, also known as HIF-1 beta), the heterodimeric complex binds TACGTG-containing genomic enhancer elements, and activates transcription of target genes. As a first step in developing genetic models to study the biology related to cellular hypoxia, we have cloned the murine HIF-1 alpha cDNA, determined the tissue-specific expression of its mRNA, functionally analyzed its protein product, and characterized its promoter and its genomic structure. A comparison between the murine and human HIF-1 alpha protein sequence reveals 95%, 99%, and 83% identity in the bHLH, PAS, and variable domains, respectively. RNAse protection assays demonstrate that in adult mice, the mHIF-1 alpha mRNA is expressed at high levels in kidney, heart, brain, thymus, and placenta, with moderate expression in liver, spleen, testis, and lung and much lower expression in skeletal muscle testis. Northern blot analysis indicates that the mRNA of the murine HIF-1 alpha is transcribed in two forms, a major 4-kb species and a minor 5-kb species; both are present in all tissues examined. The Hif-1 alpha promoter is GC rich, does not have a TATA element near its transcriptional start site, and does not respond to hypoxia or Co2+. The mHIF-1 alpha structural gene is composed of 15 exons. The splice junction sites within the bHLH and the PAS domains of HIF-1 alpha gene are highly conserved with respect to a number of previously characterized members of the bHLH-PAS superfamily. However, unlike other bHLH-PAS genes, where the variable domain is encoded by 2 exons, the variable region of the mHIF-1 alpha gene is encoded by 7 exons. Furthermore, most of these splice junction sites in the variable region are conserved with that of HIF-2 alpha, a recently cloned hypoxia-responsive bHLH-PAS protein (also known as MOP2, EPAS1, and HLF). These data suggest that HIF-1 alpha, along with HIF-2 alpha, represents a new subclass of the bHLH-PAS superfamily.

Amino Acid Sequence↗

PubMed1996

Localization of the somatostatin receptor SST2A in rat brain using a specific anti-peptide antibody.

Biological actions of somatostatin are exerted via a family of receptors, for which five genes recently have been cloned. However, none of these receptor proteins has been visualized yet in the brain. In the present-study, the regional and cellular distribution of the somatostatin sst2A receptor was investigated via immunocytochemistry in the rat central nervous system by using an antibody generated against a unique sequence of the receptor protein. Specificity of the antiserum was demonstrated by immunoblot and immunocytochemistry on rat brain membranes and/or on cells transfected with cDNA encoding the different sst receptor subtypes. In rat brain sections, sst2A receptor immunoreactivity was concentrated either in perikarya and dendrites or in axon terminals distributed throughout the neuropil. Somatodendritic labeling was most prominent in the olfactory tubercle, layers II-III of the cerebral cortex, nucleus accumbens, pyramidal cells of CA1-CA2 subfields of the hippocampus, central and cortical amygdaloid nuclei, and locus coeruleus. Labeled terminals were detected mainly in the endopiriform nucleus, deep layers of the cortex, claustrum, substantia innominata, subiculum, basolateral amygdala, medial habenula, and periaqueductal gray. Electron microscopy confirmed the association of sst2A receptors with perikarya and dendrites in the former regions and with axon terminals in the latter. These results provide the first characterization of the cellular distribution of a somatostatin receptor in mammalian brain. The widespread distribution of the sst2A receptor in cerebral cortex and limbic structures suggests that it is involved in the transduction of both pre- and postsynaptic effects of somatostatin on cognition, learning, and memory.

Animals↗

PubMed1996

Somatostatin receptor subtypes: specific expression and signaling properties.

The five cloned somatostatin (SRIF) receptors (ssts) are presumed to subserve unique biological roles by virtue of their tissue-specific expression and particular signal transduction mechanisms. However, the function of any individual sst subtype in its normal physiological milieu is not understood, because tissues and cells often express multiple ssts and, in the absence of receptor-specific SRIF analogs, the actions of individual receptors cannot be identified. To unravel the physiological role and signaling mechanism of the ssts, we have generated receptor subtype-specific antibodies and used these antibodies to determine the distribution of the receptor proteins and to identify the signal-transducing molecules with which particular sst subtypes interact.

Animals↗

PubMed1995

Function and regulation of somatostatin receptor subtypes.

The five known somatostatin receptors serve unique biological roles by virtue of their tissue-specific expression and particular biochemical properties. However, the function of any individual receptor in its normal physiological milieu is not understood. Studies to address this problem have been difficult because tissues and cell lines often express multiple somatostatin receptors and, in the absence of receptor-selective somatostatin analogues, the actions of individual receptors cannot be identified. Moreover, the biological and biochemical actions of somatostatin receptors depend on their cellular environment, so that the behaviour of a receptor expressed in heterologous cells does not necessarily mimic that of endogenous receptors. We have developed two approaches to examine somatostatin receptors which circumvent these problems. Using a biotinylated somatostatin analogue for affinity purification, we isolated somatostatin receptors together with associated G proteins. Subsequent analysis of the purified complex with G protein-specific antibodies showed that the somatostatin receptors in AR42J cells preferentially couple with two pertussis toxin-sensitive G proteins: Gi alpha 1 and Gi alpha 3. To examine individual receptor types, we developed receptor-specific antibodies and used them to show that both sstr1 and sstr2 proteins were present in the GH4C1 pituitary cell line whereas AR42J cells contained sstr2 but not sstr1. Immunoprecipitation of receptor-G protein complexes with GH4C1 cells showed that sstr1 and sstr2 are both coupled to pertussis toxin-sensitive G proteins, in contrast to the results observed when these receptors are overexpressed in some non-endocrine cells. We also showed that the somatostatin receptors in GH4C1 cells are subject to both homologous and heterologous hormonal regulation. The mechanisms involved in the regulation of different receptor types are now being characterized using the receptor-specific antibodies to isolate the individual receptor proteins. Elucidating signal transduction by endogenous somatostatin receptors as well as their hormonal regulation will be critical for understanding the functions of these receptors in the different physiological targets of somatostatin.

Animals↗

PubMed1995

Development and use of a receptor antibody to characterize the interaction between somatostatin receptor subtype 1 and G proteins.

The signal transduction pathways regulated by somatostatin receptor subtype 1 (sst1) have been difficult to define because of the variability observed when this receptor is expressed in different cell types by transfection and because pharmacological approaches are inadequate to distinguish sst1 receptor subtypes. To study the sst1 receptor in its endogenous environment, we developed a polyclonal antibody to a 15-amino acid peptide corresponding to a unique sequence in the receptor carboxyl terminus. The peptide antibody routinely precipitated 70% of the soluble [125I-Tyr11]somatostatin/receptor complex prepared from Chinese hamster ovary-K1 cells expressing the sst1 receptor but precipitated < 1% of the complex from cells expressing other sst receptor subtypes. Photoaffinity-labeled sst1 receptor was also specially immunoprecipitated and migrated as a broad 60-kDa band on sodium dodecyl sulfate polyacrylamide gels. The observation that sst receptors from GH4C1 pituitary cells were immunoprecipitated by the antibody and that receptors from AR4-2J pancreatic acinar cells were not indicated that only the former expressed sst1 receptor protein. Because reverse transcription-polymerase chain reaction showed that GH4C1 cells contained both sst1 and sst2 receptor mRNA, immunoprecipitation permitted the sst1 receptor to be separated from the other receptors present. Two observations showed that G proteins were coprecipitated with sst1 receptors from GH4C1 cells. First, pertussis toxin pretreatment markedly decreased hormone binding in the immunoprecipitate. Second, the addition of 20 microM guanosine-5'-(gamma-thio)triphosphate to the immunoprecipitated [125I-Tyr11]somatostatin/receptor complex stimulated the rate of dissociation of bound ligand by 10-fold. Interestingly, however, the dissociation rate of approximately 30% of the ligand/receptor complex was unaffected by guanosine-5'-(gamma-thio)triphosphate. In summary, we have developed an sst1 receptor-specific antibody and used it to show that sst1 receptors endogenously expressed in GH4C1 pituitary cells couple primarily to pertussis toxin-sensitive G proteins. Furthermore, these receptors exist in two distinct high affinity states distinguished by their GTP sensitivity.

Amino Acid Sequence↗

PubMed1993

Treatment of acute promyelocytic leukemia with all-trans retinoic acid. A five-year experience.

From January 1986 to April 1991, 107 consecutive patients with acute promyelocytic leukemia (APL) were treated with retinoic acid (RA) at an oral dose of 45-60mg/m2/d, alone or in combination with chemotherapy. In 91 cases treated with RA alone, 74 (81.3%) achieved complete remission (CR). The CR rate was 75% in 16 cases treated with combined therapy. Among 50 patients closely followed for a median of 36 months (4-60), 10 received RA as continuation therapy (Group A), 10 received chemotherapy (Group B) and 30 were treated with RA and chemotherapy alternately in regular sequence (Group C). The mean survival time was 8.4, 9.7 and 21.6 months, respectively, for the 29 cases who died. The survival probability was higher in Group C than in Group A and B (P < 0.01). RA did not provoke or aggravate DIC, it did not cause marrow hypoplasia or aplasia. The side effects were relatively mild as compared with chemotherapy. CFU-GM markedly reduced before treatment was restored to normal level after CR, while the result for L-CFU was reversed. In 40 cases examined for in vitro induction of differentiation, 39 responders were culminating in CR. Aberrant karyotype t (15; 17) was positive in all 47 cases examined prior to the treatment. It disappeared in all of the 20 cases studied after achieving CR, and reappeared in 3 cases following relapse. The best regimen to maintain a longer CR duration and survival time in this study was to use RA and chemotherapy alternately as continuation therapy.

Adolescent↗

PubMed1992

Construction of a genomic DNA library of the Toxoplasma gondii ZS2 strain, screening of specific clones, and DNA diagnosis of toxoplasmosis.

We have constructed a genomic DNA library of the Toxoplasma gondii ZS2 strain and isolated a specific cloned DNA sequence from this organism. The restriction map of this cloned 1.1-kb DNA fragment was analyzed. Southern and dot-blot analyses showed that the 32P-labeled DNA fragment hybridized to parasite DNA, to DNAs from peripheral blood leukocytes and the thymus of baby pigs that were artificially infected with T. gondii, and to DNAs of T. gondii-positive anencephalic and hydrocephalic fetuses. It did not hybridize with DNA from controls, (i.e., normal human and baby pig peripheral blood leukocytes, spleen of normal mice, Plasmodium falciparum, Pneumocystis carinii, and pBR322). As few as 100 T. gondii parasites or 500 pg of purified DNA from T. gondii can be detected by dot-blot hybridization. This probe method was specific and sensitive, and has been used successfully in detecting various clinical cases of toxoplasmosis with T. gondii.

Animals↗

PubMed1990

Cooperativity of sequence elements mediates tissue specificity of the rat insulin II gene.

The 5'-flanking region of the rat insulin II gene (-448 to +50) is sufficient for tissue-specific expression. To further determine the tissue-specific cis-acting element(s), important sequences defined by linker-scanning mutagenesis were placed upstream of a heterologous promoter and transfected into insulin-producing and -nonproducing cells. Rat insulin promoter element 3 (RIPE3), which spans from -125 to -86, was shown to confer beta-cell-specific expression in either orientation. However, two subregions of RIPE3, RIPE3a and RIPE3b (defined by linker-scanning mutations), displayed only marginal activities. These results suggest that the two subregions cooperate to confer tissue specificity, presumably via their cognate binding factors.

Animals↗

PubMed1990

[Construction of a genomic DNA library of Toxoplasma gondii (ZS2 strain), screening of specific clone and DNA diagnosis of toxoplasmosis].

We have constructed a genomic DNA library of Toxoplasma gondii (ZS2 strain) and screened out a specific DNA sequence for T. gondii. The restriction map of the cloned DNA fragment (1.1kb) was analysed. The Southern and dot-blot analyses showed that the 32P-labeled cloned DNA fragment hybridized to the parasite DNA, DNAs from peripheral white blood cells and thymus of baby pigs artificially infected with T. gondii and DNAs of T. gondii- positive anencephalus and hydrocephalus, but did not hybridize to DNAs from controls, i. e., normal human and baby pig peripheral white blood cells, spleen of normal mouse, Plasmodium falciparum, Pneumocystis carinii and pBR322. As few as 100 T. gondii parasites or 500pg purified DNA from T. gondii can be detected by dot blot hybridization. This established DNA probe method was specific and sensitive and has been successfully used in detecting various cases infected with T. gondii.

Animals↗

PubMed1989

Sequential observation of clinical and karyotypic evolution in a patient with myelodysplastic syndrome.

This paper reports an interesting case of myelodysplastic syndrome (MDS), whose bone marrow karyotype at diagnosis was 46, XY, t(16;17) (q12;q25). Fourteen months later, the disease transformed into erythroleukemia, and several correlative clones with hyperdiploid appeared at the same time. Thus, we consider that detecting karyotypic evolution may help evaluate the prognosis of MDS.

Anemia, Refractory↗