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Generation and Culture of Blood Outgrowth Endothelial Cells from Human Peripheral Blood

Historically, the limited availability of primary endothelial cells from patients with vascular disorders has hindered the study of the molecular ...

University of Cambridge

Phosphatidylinositol transfer proteins (PITPs) are versatile proteins required for signal transduction and membrane traffic. The best characterized mammalian PITPs are the Class I PITPs, PITPα (PITPNA) and PITPβ (PITPNB), which are single domain proteins with a hydrophobic cavity that binds a phosphatidylinositol (PI) or phosphatidylcholine molecule. In this study, we report the lipid binding properties of an uncharacterized soluble PITP, phosphatidylinositol transfer protein, cytoplasmic 1 (PITPNC1) (alternative name, RdgBβ), of the Class II family. We show that the lipid binding properties of this protein are distinct to Class I PITPs because, besides PI, RdgBβ binds and transfers phosphatidic acid (PA) but hardly binds phosphatidylcholine. RdgBβ when purified from Escherichia coli is preloaded with PA and phosphatidylglycerol. When RdgBβ was incubated with permeabilized HL60 cells, phosphatidylglycerol was released, and PA and PI were now incorporated into RdgBβ. After an increase in PA levels following activation of endogenous phospholipase D or after addition of bacterial phospholipase D, binding of PA to RdgBβ was greater at the expense of PI binding. We propose that RdgBβ, when containing PA, regulates an effector protein or can facilitate lipid transfer between membrane compartments.

Phosphatidylinositol transfer protein, cytoplasmic 1 (PITPNC1) binds and transfers phosphatidic acid.

There is growing evidence to support the use of trial-based functional analyses, particularly in classroom settings. However, there currently are no evaluations of this procedure with typically developing children. Furthermore, it is possible that refinements may be needed to adapt trial-based analyses to mainstream classrooms. This study was designed to expand the trial-based functional analysis literature by implementing the procedure in 2 mainstream primary school classrooms and validating the analysis through comparison of multiple treatment options, including some that were not indicated by the functional analysis. We also extended the procedure by including a peer-attention condition and obtaining data from teachers regarding the feasibility of the procedures. For all participants, functional analysis results helped to identify effective treatments. Furthermore, relative effects among treatments were accurately predicted by the functional analysis outcomes. Teachers reported that they understood the logic of functional analysis and found both analysis and treatment procedures to be easy and effective.

Emily Groves

Department of Medicine

University of Cambridge

Generation and Culture of Blood Outgrowth Endothelial Cells from Human Peripheral Blood

Emily Groves

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University of Cambridge

Generation and Culture of Blood Outgrowth Endothelial Cells from Human Peripheral Blood

Department of Medicine