Identification of downstream components of ubiquitin-conjugating enzyme PHOSPHATE2 by quantitative membrane proteomics in Arabidopsis roots.

MicroRNA399-mediated regulation of the ubiquitin-conjugating enzyme UBC24/phosphate2 (PHO2) is crucial for Pi acquisition and translocation in plants. Because of a potential role for PHO2 in protein degradation and its association with membranes, an iTRAQ (for isobaric tags for relative and absolute quantitation)- based quantitative membrane proteomic method was employed to search for components downstream of PHO2. A total of 7491 proteins were identified from Arabidopsis thaliana roots by mass spectrometry, 35.2% of which were predicted to contain at least one transmembrane helix. Among the quantifiable proteins, five were significantly differentially expressed between the wild type and pho2 mutant under two growth conditions. Using immunoblot analysis, we validated the upregulation of several members in phosphate transporter1 (PHT1) family and phosphate transporter traffic facilitator1 (PHF1) in pho2 and demonstrated that PHO2 mediates the degradation of PHT1 proteins. Genetic evidence that loss of PHF1 or PHT1;1 alleviated Pi toxicity in pho2 further suggests that they play roles as downstream components of PHO2. Moreover, we showed that PHO2 interacts with PHT1s in the postendoplasmic reticulum compartments and mediates the ubiquitination of endomembrane-localized PHT1;1. This study not only uncovers a mechanism by which PHO2 modulates Pi acquisition by regulating the abundance of PHT1s in the secretory pathway destined for plasma membranes, but also provides a database of the membrane proteome that will be widely applicable in root biology research.

Vacuolar Ca2+/H+ transport activity is required for systemic phosphate homeostasis involving shoot-to-root signaling in Arabidopsis.

Calcium ions (Ca(2+)) and Ca(2+)-related proteins mediate a wide array of downstream processes involved in plant responses to abiotic stresses. In Arabidopsis (Arabidopsis thaliana), disruption of the vacuolar Ca(2+)/H(+) transporters CAX1 and CAX3 causes notable alterations in the shoot ionome, including phosphate (P(i)) content. In this study, we showed that the cax1/cax3 double mutant displays an elevated P(i) level in shoots as a result of increased P(i) uptake in a miR399/PHO2-independent signaling pathway. Microarray analysis of the cax1/cax3 mutant suggests the regulatory function of CAX1 and CAX3 in suppressing the expression of a subset of shoot P(i) starvation-responsive genes, including genes encoding the PHT1;4 P(i) transporter and two SPX domain-containing proteins, SPX1 and SPX3. Moreover, although the expression of several PHT1 genes and PHT1;1/2/3 proteins is not up-regulated in the root of cax1/cax3, results from reciprocal grafting experiments indicate that the cax1/cax3 scion is responsible for high P(i) accumulation in grafted plants and that the pht1;1 rootstock is sufficient to moderately repress such P(i) accumulation. Based on these findings, we propose that CAX1 and CAX3 mediate a shoot-derived signal that modulates the activity of the root P(i) transporter system, likely in part via posttranslational regulation of PHT1;1 P(i) transporters.

Predictable capability control scheme for oxygen-exchange blood flow regulation in an extracorporeal membrane oxygenation system.

Extracorporeal membrane oxygenation system is used for rescue treatment strategies for temporary cardiopulmonary function support to facilitate adequately oxygenated blood to return into the systemic and pulmonary circulation systems. Therefore, a servo flow regulator is used to adjust the roller motor speed, while support blood flow can match the sweep gas flow (GF) in a membrane oxygenator. A generalised regression neural network is designed as an estimator to automatically estimate the desired roller pump speed and control parameters. Then, the proportional-integral-derivative controller with tuning control parameters showed good performance to achieve speed regulation and speed tracking in the desired operating point. Given the pressure of carbon dioxide, drainage blood flow, and cannula size, the proposed predictable capability control scheme can be validated to meet the intended uses in clinical applications.

Assistive technology using integrated flexible sensor and virtual alarm unit for blood leakage detection during dialysis therapy.

Blood leakages and blood loss are both serious complications during dialysis therapies. According to dialysis survey reports, these events are life-threatening issues for nephrology nurses, medical staff, and patients. When venous needle dislodgement occurs, it takes only <2.5 min of reaction time for blood loss in an adult patient, resulting in mortality. As an early-warning design, a wireless assistive technology using an integrated flexible sensor and virtual alarm unit was developed to detect blood leakage during dialysis therapies. The flexible sensor was designed using a screen print technique with printing electronic circuits on a plastic substrate. A self-organising algorithm was used to design a virtual alarm unit, consisting of a virtual direct current grid and a virtual alarm driver. In other words, this warning device was employed to identify the blood leakage levels via wireless fidelity wireless network in cloud computing. The feasibility was verified, and commercialisation designs can also be implemented in an embedded system.