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SHC and SHIP phosphorylation and interaction in response to activation of the FLT3 receptor.

The FLT3 receptor tyrosine kinase and its ligand, FL, regulate the development of hematopoietic stem cells and early B lymphoid progenitors. FL has a strong capacity to boost production of dendritic and natural killer cells in vivo, thereby providing a new and promising tool for anti-cancer immunotherapy. Intracellular FLT3 signaling involves tyrosine phosphorylation of several cytoplasmic proteins including SHC. We have found that upon FLT3 activation SHC phosphorylation occurs at tyrosine 239/240 and 313. SHC possesses two phosphotyrosine-binding domains: an amino-terminal phosphotyrosine binding (PTB) and a carboxy-terminal Src Homology 2 (SH2) domain. Neither is required for SHC phosphorylation, but the PTB domain is necessary and sufficient for SHC binding to the SH2 containing inositol phosphatase (SHIP). Overexpression of SHC increases the level of SHIP phosphorylation on tyrosines in response to FLT3 activation, suggesting that SHC availability is a limiting step for SHIP phosphorylation. This effect is observed only if the SHC PTB domain is functional. Interestingly, SHC overexpression in FLT3-activatable Ba/F3 cells limits FLT3-dependent cell growth and this effect requires tyrosine 313. Taken together, the present data show that SHC can antagonize cell proliferation induced by FLT3 stimulation and regulate phosphorylation of the SHIP negative regulator. In addition, our study provides the structural bases for SHC phosphorylation and formation of the SHC/SHIP complex.

Adaptor Proteins, Signal Transducing↗

Restoration of SHIP-1 activity in human leukemic cells modifies NF-kappaB activation pathway and cellular survival upon oxidative stress.

Nuclear factor-kappa B (NF-kappaB) is an important prosurvival transcription factor activated in response to a large array of external stimuli, including reactive oxygen species (ROS). Previous works have shown that NF-kappaB activation by ROS involved tyrosine phosphorylation of the inhibitor IkappaBalpha through an IkappaB kinase (IKK)-independent mechanism. In the present work, we investigated with more details NF-kappaB redox regulation in human leukemic cells. By using different cell lines (CEM, Jurkat and the subclone Jurkat JR), we clearly showed that NF-kappaB activation by hydrogen peroxide (H2O2) is cell-type dependent: it activates NF-kappaB through tyrosine phosphorylation of IkappaBalpha in Jurkat cells, whereas it induces an IKK-mediated IkappaBalpha phosphorylation on S32 and 36 in CEM and Jurkat JR cells. We showed that this H2O2-induced IKK activation in CEM and Jurkat JR cells is mediated by SH2-containing inositol 5'-phosphatase 1 (SHIP-1), a lipid phosphatase that is absent in Jurkat cells. Indeed, the complementation of SHIP-1 in Jurkat cells made them shift to an IKK-dependent mechanism upon oxidative stress stimulation. We also showed that Jurkat cells expressing SHIP-1 are more resistant to H2O2-induced apoptosis than the parental cells, suggesting that SHIP-1 has an important role in leukemic cell responses to ROS in terms of signal transduction pathways and apoptosis resistance, which can be of interest in improving ROS-mediated chemotherapies.

Apoptosis↗

Finding the common ground: where health and educational agendas meet--the School Health Incentive Program (SHIP) grants scheme.

ISSUE ADDRESSED: This paper investigates a school health promotion grants scheme as a strategy for increasing opportunities and support for schools to undertake a whole-school approach to health issues of their choice. METHOD: The School Health Incentive Program (SHIP) is an ongoing initiative offering annual grants and tailored support from health and education professionals. SHIP commenced in 2000 in the former South Eastern Sydney Area Health Service (SESAHS), New South Wales. It is a partnership between the Health Promotion Service (HPS), the three education sectors, parent groups, other health services and The Cancer Council NSW. RESULTS: Over the period 2000-04 SHIP awarded 103 grants to 71 schools from SESAHS, representing about 32,000 students. Average grant funding per student reached was $2.50. Schools used the Health Promoting Schools framework to plan and implement their health activities. Results included positive outcomes in the areas of curriculum planning, policy and procedure development, changes to physical and social environments and the development of partnerships between the school and the health sector, other organisations, parents and other schools. CONCLUSIONS: SHIP developed partnerships between health, education sectors and schools, encouraged schools to adopt a whole-school approach to health issues, and supported the development of subsequent health-focused initiatives.

Health Policy↗

CD22 forms a quaternary complex with SHIP, Grb2, and Shc. A pathway for regulation of B lymphocyte antigen receptor-induced calcium flux.

CD22 is a cell surface molecule that regulates signal transduction in B lymphocytes. Tyrosine-phosphorylated CD22 recruits numerous cytoplasmic effector molecules including SHP-1, a potent phosphotyrosine phosphatase that down-regulates B cell antigen receptor (BCR)- and CD19-generated signals. Paradoxically, B cells from CD22-deficient mice generate augmented intracellular calcium responses following BCR ligation, yet proliferation is decreased. To understand further the mechanisms through which CD22 regulates BCR-dependent calcium flux and proliferation, interactions between CD22 and effector molecules involved in these processes were assessed. The adapter proteins Grb2 and Shc were found to interact with distinct and specific regions of the CD22 cytoplasmic domain. Src homology-2 domain-containing inositol polyphosphate-5'-phosphatase (SHIP) also bound phosphorylated CD22, but binding required an intact CD22 cytoplasmic domain. All three molecules were bound to CD22 when isolated from BCR-stimulated splenic B cells, indicating the formation of a CD22.Grb2.Shc.SHIP quaternary complex. Therefore, SHIP associating with CD22 may be important for SHIP recruitment to the cell surface where it negatively regulates calcium influx. Although augmented calcium responses in CD22-deficient mice should facilitate enhanced c-Jun N-terminal kinase (JNK) activation, BCR ligation did not induce JNK activation in CD22-deficient B cells. These data demonstrate that CD22 functions as a molecular "scaffold" that specifically coordinates the docking of multiple effector molecules, in addition to SHP-1, in a context necessary for BCR-dependent SHIP activity and JNK stimulation.

Adaptor Proteins, Signal Transducing↗

SHIP is a negative regulator of growth factor receptor-mediated PKB/Akt activation and myeloid cell survival.

SHIP is an inositol 5' phosphatase that hydrolyzes the PI3'K product PI(3,4,5)P3. We show that SHIP-deficient mice exhibit dramatic chronic hyperplasia of myeloid cells resulting in splenomegaly, lymphadenopathy, and myeloid infiltration of vital organs. Neutrophils and bone marrow-derived mast cells from SHIP-/- mice are less susceptible to programmed cell death induced by various apoptotic stimuli or by growth factor withdrawal. Engagement of IL3-R and GM-CSF-R in these cells leads to increased and prolonged PI3'K-dependent PI(3,4,5)P3 accumulation and PKB activation. These data indicate that SHIP is a negative regulator of growth factor-mediated PKB activation and myeloid cell survival.

Animals↗

Effect on graft and patient survival between shipped and locally transplanted well-matched cadaveric renal allografts in Australia over a 10-year period.

BACKGROUND: The national sharing of well-matched cadaveric renal allografts has been established since 1992 in Australia. It remains uncertain whether the shipment of renal allografts interstate has a detrimental effect on graft and patient survival. METHODS: Using data collected by the Australia and New Zealand Dialysis and Transplant Registry (ANZDATA), graft and patient survival were compared between shipped (n = 623) and locally transplanted allografts (n = 894) in Australia between January 1992 and December 2001. RESULTS: Shipped renal allografts were from younger donors and had a longer ischaemic time, and were transplanted into higher risk recipients. Shipping of allografts had no effect on graft or patient survival, acute rejection at 6 months, presence of delayed graft function and serum creatinine at 6 and 12 months. In addition, human leucocyte antigen-matching and allograft ischaemic time had no effect on graft or patient survival. CONCLUSION: Recipients of either shipped or locally transplanted well-matched renal allografts have similar graft and patient survivals.

Adult↗

In vivo efficacy of shipped HLA-matched platelets.

BACKGROUND: Platelet (PLT) concentrates are currently stored in an incubator at 20 to 24 degrees C with continuous gentle agitation. PLTs are routinely shipped for transfusion to thrombocytopenic patients, however. There is a concern that PLT concentrates may be adversely affected during the shipping process. CASE REPORT: A 40-year-old woman with severe aplastic anemia and immune refractory to unselected PLT transfusions was transferred to a distant medical center for a hematopoietic peripheral blood progenitor cell transplant where she continued to receive HLA-matched PLTs from her dedicated donors. Sixteen such components were collected and air-shipped in insulated boxes to the transplant center. Thirty-seven plateletpheresis components from the same dedicated donors had been transfused to the patient before transfer. Corrected count increments (CCIs) at the two sites were compared, with assessment of the role of HLA-match grades. The mean interruption time of controlled agitation during shipment was approximately 10.5 hours. The mean CCI of all distant transfusions was 14,450 +/- 9700 PLTs per microL x m2 per 10(11) and that of local transfusions was 10730 +/- 4870. The mean donor-paired difference between CCIs at the two sites was 1140 +/- 9940. At the remote location no clinically significant bleeding occurred and one posttransfusion febrile reaction was noted. CONCLUSION: Despite the study limitations, the effectiveness, in a single patient, of leukoreduced, irradiated apheresis PLTs shipped by lengthy combined surface and airline transport is reported, as measured by posttransfusion CCIs.

Adult↗

Geoacoustic inversion in time domain using ship of opportunity noise recorded on a horizontal towed array.

A time domain geoacoustic inversion method using ship noise received on a towed horizontal array is presented. The received signal, containing ship noise as a source of opportunity, is time-reversed and then back-propagated to the vicinity of the ship. The back-propagated signal is correlated with the modeled signal which is expected to peak at the ship's location in case of a good match for the environment. This match is utilized for the geoacoustic parameter inversion. The objective function for this optimization problem is thus defined as the normalized power focused in an area around the source position, using a matched impulse response filter. A hybrid use of global and local search algorithms, i.e., GA and Powell's method is applied to the optimization problem. Applications of the proposed inversion method to MAPEX 2000 noise experiment conducted north of the island of Elba show promising results, and it is shown that the time domain inversion takes advantage of dominant frequencies of the source signature automatically.

Journal Article↗

Radiated noise characteristics of a modern cargo ship

Extensive measurements were made of the radiated noise of M/V OVERSEAS HARRIETTE, a bulk cargo ship (length 173 m, displacement 25 515 tons) powered by a direct-drive low-speed diesel engine-a design representative of many modern merchant ships. The radiated noise data show high-level tonal frequencies from the ship's service diesel generator, main engine firing rate, and blade rate harmonics due to propeller cavitation. Radiated noise directionality measurements indicate that the radiation is generally dipole in form at lower frequencies, as expected. There are some departures from this pattern that may indicate hull interactions. Blade rate source level (174 dB re 1 microPa/m at 9 Hz, 16 knots) agrees reasonably well with a model of fundamental blade rate radiation previously reported by Gray and Greeley, but agreement for blade rate harmonics is not as good. Noise from merchant ships elevates the natural ambient by 20-30 dB in many areas; the effects of this noise on the biological environment have not been widely investigated.

Journal Article↗

Influence of SHIP on the NK repertoire and allogeneic bone marrow transplantation.

Natural killer cell (NK) receptors for major histocompatibility complex (MHC) class I influence engraftment and graft-versus-tumor effects after allogeneic bone marrow transplantation. We find that SH2-containing inositol phosphatase (SHIP) influences the repertoire of NK receptors. In adult SHIP-/- mice, the NK compartment is dominated by cells that express two inhibitory receptors capable of binding either self or allogeneic MHC ligands. This promiscuous repertoire has significant functional consequences, because SHIP-/- mice fail to reject fully mismatched allogeneic marrow grafts and show enhanced survival after such transplants. Thus, SHIP plays an important role in two processes that limit the success of allogeneic marrow transplantation: graft rejection and graft-versus-host disease.

Animals↗

The effect of phosphatases SHP-1 and SHIP-1 on signaling by the ITIM- and ITAM-containing Fcgamma receptors FcgammaRIIB and FcgammaRIIA.

Inositol and tyrosine phosphatases have been implicated in inhibitory signaling by an Fc receptor for immunoglobulin G, FcgammaRIIB, in B cells, mast cells, and monocytes. Here, we propose a role for the Src homology 2 (SH2)-containing tyrosine phosphatase-1 (SHP-1) in FcgammaRIIB-mediated inhibition of FcgammaR signaling. Coexpression of SHP-1 enhances FcgammaRIIB-mediated inhibition of FcgammaRIIA phagocytosis in COS-1 cells. SHP-1 also enhances the reduction in FcgammaRIIA tyrosine phosphorylation that accompanies this inhibition. Significantly, tyrosine phosphorylation of Syk kinase is substantially inhibited by SHP-1. Furthermore, the activation of SHP-1 tyrosine phosphorylation is observed following stimulation of FcgammaRII in COS-1 cells and in human monocytes. The SH2 domain containing inositol phosphatase (SHIP), SHIP-1 also enhances FcgammaRIIB-mediated inhibition of FcgammaRIIA, indicating that FcgammaRIIB can use more than one pathway for its inhibitory action. In addition, SHP-1 and SHIP-1 can inhibit FcgammaRIIA phagocytosis and signal transduction in the absence of FcgammaRIIB. The data support emerging evidence that SH2-containing phosphatases, such as SHP-1 and SHIP-1, can modulate signaling by "activating" receptors.

Amino Acid Motifs↗

Loss of SHIP and CIS recruitment to the granulocyte colony-stimulating factor receptor contribute to hyperproliferative responses in severe congenital neutropenia/acute myelogenous leukemia.

Mutations in the G-CSF receptor (G-CSFR) in patients with severe congenital neutropenia (SCN) are postulated to contribute to transformation to acute myelogenous leukemia (AML). These mutations result in defective receptor internalization and sustained cellular activation, suggesting a loss of negative signaling by the G-CSFR. In this paper we investigated the roles of SHIP and cytokine-inducible Src homology 2 protein (CIS) in down-modulating G-CSFR signals and demonstrate that loss of their recruitment as a consequence of receptor mutations leads to aberrant signaling. We show that SHIP binds to phosphopeptides corresponding to Tyr744 and Tyr764 in the G-CSFR and that Tyr764 is required for in vivo phosphorylation of SHIP and the formation of SHIP/Shc complexes. Cells expressing a G-CSFR form lacking Tyr764 exhibited hypersensitivity to G-CSF and enhanced proliferation, but to a lesser degree than observed with the most common mutant G-CSFR form in patients with SCN/AML, prompting us to investigate whether suppressor of cytokine signaling proteins also down-modulate G-CSFR signals. G-CSF was found to induce the expression of CIS and of CIS bound to phosphopeptides corresponding to Tyr729 and Tyr744 of the G-CSFR. The expression of CIS was prolonged in cells with the SCN/AML mutant G-CSFR lacking Tyr729 and Tyr744, which also correlated with increased G-CSFR expression. These findings suggest that SHIP and CIS interact with distal phosphotyrosine residues in the G-CSFR to negatively regulate G-CSFR signaling by limiting proliferation and modulating surface expression of the G-CSFR, respectively. Novel therapeutic approaches targeting inhibitory pathways that limit G-CSFR signaling may have promise in the treatment of patients with SCN/AML.

Carrier Proteins↗

Nonspecific desensitization, functional memory, and the characteristics of SHIP phosphorylation following IgE-mediated stimulation of human basophils.

Previous studies of secretion from basophils have demonstrated the phenomenon called nonspecific desensitization, the ability of one IgE-mediated stimulus to alter the cell's response to other non-cross-reacting IgE-mediated stimuli, and a process that would modify phosphatidylinositol 3,4,5-phosphate levels was speculated to be responsible for nonspecific desensitization. The current studies examined the changes and characteristics of SHIP1 phosphorylation as a measure of SHIP1 participation in the reaction. Based on the earlier studies, two predictions were made that were not observed. First, the kinetics of SHIP1 phosphorylation were similar to reaction kinetics of other early signals and returned to resting levels while nonspecific desensitization remained. Second, in contrast to an expected exaggerated SHIP phosphorylation, cells in a state of nonspecific desensitization showed reduced SHIP phosphorylation (compared with cells not previously exposed to a non-cross-reacting Ag). Discordant with expectations concerning partial recovery from nonspecific desensitization, treatment of cells with DNP-lysine to dissociate bound DNP-HSA, either enhanced or had no effect on SHIP phosphorylation following a second Ag. These experiments also showed a form of desensitization that persisted despite dissociation of the desensitizing Ag. Recent studies and the results of these studies suggest that loss of early signaling components like syk kinase may account for some of the effects of nonspecific desensitization and result in a form of immunological memory of prior stimulation. Taken together, the various characteristics of SHIP phosphorylation were not consistent with expectations for a signaling element involved in nonspecific desensitization, but instead one which itself undergoes nonspecific desensitization.

Antibodies, Anti-Idiotypic↗

[Study of mutation and single nucleotide polymorphism of PDGFRbeta and SHIP gene in acute myeloid leukemia].

OBJECTIVE: To investigate the significance of mutation and single nucleotide polymorphism (SNP) of class III receptor tyrosine kinases such as PDGFRbeta and SHIP in acute myeloid leukemia (AML) patients. METHODS: Screening of the mutation and SNP of PDGFRbeta and SHIP by genomic PCR, RT-PCR, directly sequencing and Mass-ARRAY system was carried out in 273 AML patients. RESULTS: The mutations of PDGFRbeta R685C and SHIP Q1153L were detected for the first time in AML patients. The positivity ratio was 0.73% and 0.36% respectively. CONCLUSION: The mutations of PDGFRbeta R685C and SHIP Q1153L may contribute to leukemogenesis of AML.

Humans↗

Physiological stabilization of rabbits after shipping.

Significant physiological variations that could influence experimental outcomes have been described in laboratory animals following shipping. The objective of the present study was to monitor a variety of physiologic parameters in rabbits after shipping, and to evaluate the time necessary for stabilization of these variables in the new environment. Data indicate that rabbits develop anorexia, hyperglycemia, neutrophilia, lymphopenia and elevated plasma cortisol concentrations immediately after shipping. Most of these effects abate within 2 days after arrival, suggesting that a minimum stabilization period of 48 hours after shipping is advisable prior to use of rabbits in experimental paradigms.

Aircraft↗

Lead exposure of welders and bystanders in a ship repair yard.

Blood samples were collected from 126 long-term ship repair workers and 42 retirees. Lead concentrations were determined by electrothermal atomic absorption spectrophotometry. Three groups with different degrees of lead exposure were identified: (1) 59 welders, burners, and combination men (median blood lead, 39 micrograms/100 ml); (2) 67 painters, laborers, shipfitters, riggers, and other ship repair workers (median blood lead, 26 micrograms/100 ml); and (3) 42 retired welders, painters, and shipfitters (median blood lead, 23 micrograms/100 ml). Thirty active ship repair workers, including 28 welders and burners, had blood lead concentrations above the OSHA "action level" of 40 micrograms/100 ml. Increased levels in several men without direct exposures to lead were apparently caused by "bystander's exposure." In the retirees, blood lead levels decreased with increasing length of retirement. Thus, after eight or more years, the average level was below 20 micrograms/100 ml.

Adult↗

Distribution, enrichment and accumulation of heavy metals in coastal sediments of Alang-Sosiya ship scrapping yard, India.

Since its inception in 1982, the Alang-Sosiya yard has become the largest ship scrapping works in the world. Several hundreds of ships arrive every year. The degree of heavy metal contamination has been studied in bulk and fine sediments from the intertidal zone of this ship scrapping yard, two stations, one on either side at 5 km distance and one reference station 60 km distance near Mahuva, towards the south. The samples have been subjected to a total digestion technique and analysed for elements: Cd, Co, Cu, Cr, Fe, Mn, Ni, Pb, Zn and Al, and %TOC. The absolute metal concentrations reflected variations in BF and FF sediment samples with organic matter content. Enrichment factors (EF) and geoaccumulation indices (Igeo) have been calculated and the relative contamination levels are assessed at these sites. At Alang-Sosiya, the enrichment of heavy metals has been observed to be relatively high.

Environmental Monitoring↗

The significance of dilution in evaluating possible impacts of wastewater discharges from large cruise ships.

In response to public concerns about discharges from large cruise ships, Alaska's Department of Environmental Conservation (ADEC) sampled numerous effluents in the summer of 2000. The data showed that basic marine sanitation device (MSD) technology for black water (sewage) was not performing as expected. Untreated gray water had high levels of conventional pollutants and surprisingly high levels of bacteria. Both black water and gray water discharges sometimes exceeded state water quality standards for toxicants. The state convened a Science Advisory Panel (the Panel) to evaluate impacts associated with cruise ship wastewater discharges. The effluent data received wide media coverage and increased public concerns. Consequently, legislative decisions were made at the State and Federal level, and regulations were imposed before the Panel completed its evaluation. The Panel demonstrated that following the rapid dilution from moving cruise ships, the effluent data from the Summer of 2000 would not have exceeded water quality standards, and environmental effects were not expected.

Alaska↗