Vascular and neoplastic risk in a large cohort of patients with polycythemia vera.

Purpose: The clinical course of polycythemia vera is often complicated by thrombosis as well as by the possible transition to myeloid metaplasia with myelofibrosis or acute myeloid leukemia. The aim of this study was to assess the rate of these complications in subjects receiving currently recommended treatments.

Patients And Methods: Overall, 1,638 patients from 12 countries were enrolled onto a large, prospective multicenter project aimed at describing the clinical history of polycythemia vera for the following outcomes: survival, the cumulative rate of cardiovascular death and thrombosis, the cumulative rate of leukemia, myelodysplasia, and myelofibrosis.

Constitutive mobilization of CD34+ cells into the peripheral blood in idiopathic myelofibrosis may be due to the action of a number of proteases.

Idiopathic myelofibrosis (IM) is characterized by increased numbers of CD34(+) cells in the peripheral blood (PB). We explored the possible mechanisms underlying this abnormal trafficking of CD34(+) cells. Plasma levels of neutrophil elastase (NE), total and active matrix metalloproteinase 9 (MMP-9), and soluble vascular cell adhesion molecule-1 (sVCAM-1) were dramatically increased in IM.

Cytochrome b6 arginine 214 of Synechococcus sp. PCC 7002, a key residue for quinone-reductase site function and turnover of the cytochrome bf complex.

Quinone-reductase (Q(i)) domains of cyanobacterial/chloroplast cytochrome bf and bacterial/mitochondrial bc complexes differ markedly, and the cytochrome bf Q(i) site mechanism remains largely enigmatic. To investigate the bf Q(i) domain, we constructed the mutation R214H, which substitutes histidine for a conserved arginine in the cytochrome b(6) polypeptide of the cyanobacterium Synechococcus sp. SPCC 7002.

Acute leukemia in polycythemia vera: an analysis of 1638 patients enrolled in a prospective observational study.

Progression to acute myeloid leukemia/myelodysplastic syndromes (AML/MDS) is a possible evolution of polycythemia vera (PV), but whether some patients are at increased natural risk for this complication and how much the contribution of pharmacologic cytoreduction can affect the natural course of the disease remain uncertain. The European Collaboration on Low-dose Aspirin in Polycythemia Vera (ECLAP) prospective project included 1638 patients with PV. AML/MDS was diagnosed in 22 patients after a median of 2.

The constitutive mobilization of bone marrow-repopulating cells into the peripheral blood in idiopathic myelofibrosis.

Idiopathic myelofibrosis (IM) is characterized by the constitutive mobilization of CD34(+) cells. IM peripheral blood (PB) CD34(+) cells had a reduced cloning efficiency and a lower frequency of cobblestone areas compared with normal granulocyte colony-stimulating factor (G-CSF)-mobilized PB CD34(+) cells. IM CD34(+) cells engrafted nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice, demonstrating that they contain bone marrow (BM)-repopulating cells.

The central role of the green alga Chlamydomonas reinhardtii in revealing the mechanism of state transitions.

This review focuses on the essential role played by the green alga Chlamydomonas reinhardtii in revealing both the mechanism and the physiological consequences of state transitions. Two aspects are considered. The first is the role of the cytochrome b6f complex in regulating state transitions, in light of the recently obtained 3D structure.

Reduction of the thylakoid electron transport chain by stromal reductants--evidence for activation of cyclic electron transport upon dark adaptation or under drought.

The reduction of P700(+), the primary electron donor of photosystem I (PSI), following a saturating flash of white light in the presence of the photosystem II (PSII) inhibitor 3-(3.4-dichlorophenyl)-1,1-dimethylurea (DCMU), was examined in barley plants exposed to a variety of conditions. The decay kinetic fitted to a double exponential decay curve, implying the presence of two distinct pools of PSI.

The chloroplast Rieske iron-sulfur protein. At the crossroad of electron transport and signal transduction.

We have addressed the functional and structural roles of three domains of the chloroplast Rieske iron-sulfur protein; that is, the flexible hinge that connects the transmembrane helix to the soluble cluster-bearing domain, the N-terminal stromal protruding domain, and the transmembrane helix. To this aim mutants were generated in the green alga Chlamydomonas reinhardtii. Their capacities to assemble the cytochrome b6f complex, perform plastoquinol oxidation, and signal redox-induced activation of the light-harvesting complex II kinase during state transition were tested in vivo.

A zeaxanthin-independent nonphotochemical quenching mechanism localized in the photosystem II core complex.

Illumination of dark-adapted barley plants with low light transiently induced a large nonphotochemical quenching of chlorophyll fluorescence. This reaction was identified as a form of high-energy-state quenching. Its appearance was not accompanied by zeaxanthin synthesis but was associated with a reversible inactivation of a fraction of photosystem II (PSII) centers.

A prospective analysis of thrombotic events in the European collaboration study on low-dose aspirin in polycythemia (ECLAP).

Background: - The clinical course of polycythemia vera (PV) is often complicated by arterial and venous thrombosis. Current information on these vascular complications derives from very few prospective clinical trials and several retrospective studies. The purpose of this analysis is to report the incidence, clinical features and risk factors for thrombosis in a large number of patients prospectively followed in a multicountry European study.

Chronic myeloproliferative disorders.

The Philadelphia chromosome-negative chronic myeloproliferative disorders (CMPD), polycythemia vera (PV), essential thrombocythemia (ET) and chronic idiopathic myelofibrosis (IMF), have overlapping clinical features but exhibit different natural histories and different therapeutic requirements. Phenotypic mimicry amongst these disorders and between them and nonclonal hematopoietic disorders, lack of clonal diagnostic markers, lack of understanding of their molecular basis and paucity of controlled, prospective therapeutic trials have made the diagnosis and management of PV, ET and IMF difficult. In Section I, Dr.

Treatment indications and choice of a platelet-lowering agent in essential thrombocythemia.

The therapeutic strategy in patients with essential thrombocythemia (ET) is a difficult balance between the prevention of bleeding and thrombotic complications and the risks of drug side effects and toxicity. Major bleeding is rare and seems to be related to higher platelet counts. Therefore, a platelet count greater than 1500 x 10(9)/L is generally regarded as an indication for cytoreduction.

From light to life: an interdisciplinary journey into photosynthetic activity.

Workshop on Molecular Genetics and Biophysical Aspects of Photosynthesis

In vivo changes of the oxidation-reduction state of NADP and of the ATP/ADP cellular ratio linked to the photosynthetic activity in Chlamydomonas reinhardtii.

The ATP/ADP and NADP/NADPH ratios have been measured in whole-cell extract of the green alga Chlamydomonas reinhardtii, to understand their availability for CO(2) assimilation by the Calvin cycle in vivo. Measurements were performed during the dark-light transition of both aerobic and anaerobic cells, under illumination with saturating or low light intensity. Two different patterns of behavior were observed: (a) In anaerobic cells, during the lag preceding O(2) evolution, ATP was synthesized without changes in the NADP/NADPH ratio, consistently with the operation of cyclic electron flow.

Plastocyanin is indispensable for photosynthetic electron flow in Arabidopsis thaliana.

Plastocyanin is a soluble copper-containing protein present in the thylakoid lumen, which transfers electrons to photosystem I. In the chloroplast of the flowering plant Arabidopsis thaliana, a cytochrome c6-like protein is present, which was recently suggested to function as an alternative electron carrier to plastocyanin. We show that Arabidopsis plants mutated in both of the two plastocyanin-coding genes and with a functional cytochrome c6 cannot grow photoautotrophically because of a complete block in light-driven electron transport.

Thylakoid targeting of Tat passenger proteins shows no delta pH dependence in vivo.

The Tat pathway is a major route for protein export in prokaryotes and for protein targeting to thylakoids in chloroplasts. Based on in vitro studies, protein translocation through this pathway is thought to be strictly dependent on a transmembrane delta pH. In this paper, we assess the delta pH sensitivity of the Tat pathway in vivo.

Redox-coupled proton pumping activity in cytochrome b6f, as evidenced by the pH dependence of electron transfer in whole cells of Chlamydomonas reinhardtii.

The pH dependence of cytochrome b(6)f catalytic activity has been measured in whole cells of the green alga Chlamydomonas reinhardtii over the 5-8 range. An acid pH slowed the reactions occurring at the lumenal side of the complex (cytochrome b(6) and f reduction) and affected also the rate and amplitude of the slow electrogenic reaction (phase b), which is supposed to reflect transmembrane electron flow in the complex. On the other hand, a direct measurement of the transmembrane electron flow from the kinetics of cytochrome b(6) oxidation revealed no pH sensitivity.

Diagnosis of Mycobacterium bovis infection in calves sensitized by mycobacteria of the avium/intracellulare group.

Because of the frequent exposure of cattle to mycobacteria of the avium/intracellulare group, an investigation was carried out into the possible repercussions thereof on the diagnosis of bovine tuberculosis. Three calves from a bovine tuberculosis-free herd, scored avian reactors in the gamma-interferon assay for bovine tuberculosis, were sedated and inoculated endotracheally with a virulent Mycobacterium bovis strain. Then, three other avian reactors were housed with the above donor calves.

Involvement of state transitions in the switch between linear and cyclic electron flow in Chlamydomonas reinhardtii.

The energetic metabolism of photosynthetic organisms is profoundly influenced by state transitions and cyclic electron flow around photosystem I. The former involve a reversible redistribution of the light-harvesting antenna between photosystem I and photosystem II and optimize light energy utilization in photosynthesis whereas the latter process modulates the photosynthetic yield. We have used the wild-type and three mutant strains of the green alga Chlamydomonas reinhardtii--locked in state I (stt7), lacking the photosystem II outer antennae (bf4) or accumulating low amounts of cytochrome b6f complex (A-AUU)--and measured electron flow though the cytochrome b6f complex, oxygen evolution rates and fluorescence emission during state transitions.

Chimeric fusions of subunit IV and PetL in the b6f complex of Chlamydomonas reinhardtii: structural implications and consequences on state transitions.

The cytochrome b(6)f complex of Chlamydomonas reinhardtii contains four large subunits and at least three small ones, PetG, PetL, and PetM, whose role and location are unknown. Chimeric proteins have been constructed, in which the C terminus of subunit IV is fused to either one or the other of the two putative N termini of PetL. Biochemical and functional analysis of the chimeras together with mass spectrometry analysis of the wild-type (WT) complex led to the following conclusions: (i) neither a free subunit IV C terminus nor a free PetL N terminus is required for assembly of the b(6)f complex; (ii) the first AUG codon in the sequence of the gene petL is used for initiation; (iii) the N terminus of WT PetL lies in the lumen; (iv) in the WT complex, the N terminus of PetL and the C terminus of subunit IV are within reach of each other; (v) the purified b(6)f complex from C.

A Polycythemia Vera Updated: Diagnosis, Pathobiology, and Treatment.

This review focuses on polycythemia vera (PV)-its diagnosis, cellular and genetic pathology, and management. In Section I, Dr. Pearson, with Drs.

The epidemiology of the antiphospholipid syndrome: who is at risk?

Arterial and venous thrombosis are the most common and clinically relevant events of the so-called antiphospholipid syndrome; they are reported in approximately one third of patients with the antiphospholipid (aPL) antibodies. APL antibodies are part of a wide family of immunoglobulins directed against proteins complexed with negatively charged phospholipids. They include lupus anticoagulants (LA), anticardiolipin (aCL) antibodies, and the most recently recognized anti-beta-2-glycoprotein I (beta 2-GPI) and antiprothrombin (aPT) antibodies.