Phosphoproteomic analysis of chromoplasts from sweet orange during fruit ripening

Like other types of plastids, chromoplasts have essential biosynthetic and metabolic activities which may be regulated via post‐translational modifications, such as phosphorylation, of their resident proteins. We here report a proteome‐wide mapping of in vivo phosphorylation sites in chromoplast‐enriched samples prepared from sweet orange [Citrus sinensis (L.) Osbeck] at different ripening stages by titanium dioxide‐based affinity chromatography for phosphoprotein enrichment with LC‐MS/MS. A total of 109 plastid‐localized phosphoprotein candidates were identified that correspond to 179 unique phosphorylation sites in 135 phosphopeptides. On the basis of Motif‐X analysis, two distinct types of phosphorylation sites, one as proline‐directed phosphorylation motif and the other as casein kinase II motif, can be generalized from these identified phosphopeptides. While most identified phosphoproteins show high homology to those already identified in plastids, approximately 22% of them are novel based on BLAST search using the public databases PhosPhAt and P³DB. A close comparative analysis showed that approximately 50% of the phosphoproteins identified in citrus chromoplasts find obvious counterparts in the chloroplast phosphoproteome, suggesting a rather high‐level of conservation in basic metabolic activities in these two types of plastids. Not surprisingly, the phosphoproteome of citrus chromoplasts is also characterized by the lack of phosphoproteins involved in photosynthesis and by the presence of more phosphoproteins implicated in stress/redox responses. This study presents the first comprehensive phosphoproteomic analysis of chromoplasts and may help to understand how phosphorylation regulates differentiation of citrus chromoplasts during fruit ripening.     

IKK�� downregulation is critical for triggering JNKs-dependent cell apoptotic response in the human hepatoma cells under arsenite exposure

Arsenite has a long history in treating leukemia, which might be also effective in the therapy of other cancers. Our previous published data have demonstrated that arsenite exposure induces apoptosis in the HepG2 human hepatoma cells via activating JNKs/AP-1 pathway, but the upstream signaling events responsible for JNKs (c-Jun N-terminal kinase) cascade activation have not been fully discovered. Since cross-talk between IKK/NF-κB and JNKs pathways under stress conditions is a hot topic, in this article, we investigate the potential roles of IKKα and IKKβ, the catalytic subunits of IKK complexes, in the arsenite-induced JNKs pathway activation in the HepG2 cells. We found that arsenite exposure induced JNKs and AP-1 activation accompanying with a significant reduction of both IKKα and IKKβ expressions. Overexpression of IKKβ, but not of IKKα, inhibited arsenite-induced MKK7/JNKs/AP-1 pathway activation as well as the apoptotic response. Therefore, we conclude that the downregulation of IKKβ expression is the prerequisite signaling event for mediating JNKs pathway activation and the cellular apoptotic response in the HepG2 cells under arsenite exposure. Targeting IKKβ might be helpful to enhance the tumor therapeutic effect of arsenite.     

Overexpression of IL-10 in C2D Macrophages Promotes a Macrophage Phenotypic Switch in Adipose Tissue Environments

Adipose tissue macrophages are a heterogeneous collection of classically activated (M1) and alternatively activated (M2) macrophages. Interleukin 10 (IL-10) is an anti-inflammatory cytokine, secreted by a variety of cell types including M2 macrophages. We generated a macrophage cell line stably overexpressing IL-10 (C2D-IL10) and analyzed the C2D-IL10 cells for several macrophage markers after exposure to adipocytes compared to C2D cells transfected with an empty vector (C2D-vector). C2D-IL10 macrophage cells expressed more CD206 when co-cultured with adipocytes than C2D-vector cells; while the co-cultured cell mixture also expressed higher levels of Il4, Il10, Il1β and Tnf. Since regular C2D cells traffic to adipose tissue after adoptive transfer, we explored the impact of constitutive IL-10 expression on C2D-IL10 macrophages in adipose tissue in vivo. Adipose tissue-isolated C2D-IL10 cells increased the percentage of CD206⁺, CD301⁺, CD11c⁻CD206⁺ (M2) and CD11c⁺CD206⁺ (M1b) on their cell surface, compared to isolated C2D-vector cells. These data suggest that the expression of IL-10 remains stable, alters the C2D-IL10 macrophage cell surface phenotype and may play a role in regulating macrophage interactions with the adipose tissue.     

The Optimal Velocity Criterion in the Diagnosis of Unilateral Middle Cerebral Artery Stenosis by Transcranial Doppler

We evaluated the optimal flow velocity of transcranial doppler (TCD) in detecting unilateral middle cerebral artery (MCA) stenosis and stenosis grading by magnetic resonance angiography (MRA) as the reference standard. 302 nonconsecutive patients with unilateral MCA stenosis detected by TCD underwent MRA of the intracranial arteries. The peak systolic velocity (PSV), mean flow velocity (MFV), and end-diastolic velocity (EDV) of each MCA were recorded. 604 MCA were categorized into four groups depending on the stenosis severity: normal MCA (n = 319, 52.8 %), mild stenosis (n = 94, 15.6 %), moderate stenosis (n = 66, 10.9 %), and severe stenosis (n = 125, 20.7 %). Significant differences in PSV, MFV, and EDV between these four groups were observed (P < 0.001, respectively). The optimal cutoff velocities for detecting MCA stenosis were: PSV = 160 cm/s, MFV = 100 cm/s, EDV = 60 cm/s; the optimal cutoff points to distinguish mild from moderate stenosis were: PSV = 200 cm/s, MFV = 120 cm/s, EDV = 80 cm/s; the cutoffs to distinguish moderate from severe stenosis were: PSV = 280 cm/s, MFV = 180 cm/s, EDV = 110 cm/s. Using PSV as the diagnostic criteria, the correlation for diagnosing MCA stenosis using TCD and MCA was good (Kappa number κ = 0.668); using as MFV criteria, κ = 0.641. The optimal cutoff PSV values in stenosis grading on TCD were 160, 200, and 280 cm/s. The optimal cutoff MFV values were 100, 120, and 180 cm/s. PSV is more accurate than MFV in detecting and grading MCA stenosis.     

苜蓿与沙打旺苗期生长和水分利用对土壤水分变化的反应

通过室内生长箱内盆栽实验,比较了苜蓿和沙打旺苗期的根冠生长和水分利用对种土壤水分环境变化的响应和差异.结果表明,充分供水下苜蓿和沙打旺苗期生物量和蒸腾效率均最高,苜蓿均显著高于沙打旺.土壤水分减少后苜蓿苗期生物量和蒸腾效率下降幅度均大于沙打旺.从低水到阶段低水处理后土壤水分逐渐降低和降低后再复水到低水处理,苜蓿和沙打旺的生物量分别较持续低水处理显著减少47.8%和27.9%.旱后复水后苜蓿根冠比和单位根量耗水量较显著增加,蒸腾效率显著下降; 沙打旺根冠比显著下降,单位根量耗水量和蒸腾效率变化不显著.