Final Technical Report: Genetic and Molecular Analysis of a new control pathway in assimilate partitioning.

Assimilate partitioning refers to the systemic distribution of photoassimilate from sites of primary assimilation (source tissue) to import-dependent tissues and organs (sinks). One of the defining questions in this area is how plants balance source productivity with sink demand. We discovered a sucrose-sensing signal transduction pathway that controls the activity of BvSUT1, a proton-sucrose symporter in sugar beet leaf tissue. Sucrose symporters are responsible for sucrose accumulation in the phloem of many plants and, therefore, they mediate the pivotal step in the long-distance transport of photoassimilate to non-photosynthetic tissues, such as roots and seed. We previously showed that sucrose transport activity is directly proportional to the transcription rate of BvSUT1 and that symporter mRNA and protein have high rates of turnover with half-lives on the order of 2 h. We further demonstrated that symporter transcription is regulated by sucrose levels in the leaf and that sucrose-dependent regulation of BvSUT1 transcription is mediated, at least in part, by a protein phosphorylation relay pathway. The goal of the experiments during this current grant were to use genetic and molecular approaches to identify essential components of this vital regulatory system. The initial objectives were to: (1) to characterize Arabidopsis mutants we've isolated that are resistant to growth inhibition by sucrose analogues that are recognized by the sucrose-sensor, (2) to screen for loss of function mutants in BvSUT1-promoter:luciferase transgenic plants that no longer respond to sucrose accumulation in the leaf using non-destructive visualization of luciferase activity, (3) to use gel mobility-shift assays and nuclease protection experiments to identify cis elements in the symporter promoter and DNA-binding proteins that are involved in sucrose regulation of symporter expression

The Uranium-Trend Dating Method: Principles and Application for Southern California Marine Terrace Deposits

Uranium-trend dating is an open-system method for age estimation of Quaternary sediments, using disequilibrium in the 238U–234U–230Th decay series. The technique has been applied to alluvium, colluvium, loess, till, and marine sediments, in this study we tested the U-trend dating method on calcareous marine terrace deposits from the Palos Verdes Hills and San Nicolas Island, California. Independent age estimates indicate that terraces in these areas range from –80 ka to greater than 1.0 Ma. Two low terraces on San Nicolas Island yielded U-trend plots that have a clustered array of points and the ages of these deposits are indeterminate or highly suspect. Middle Pleistocene terraces and one early Pleistocene terrace on San Nicolas Island and all terraces on the Palos Verdes Hills gave reasonably linear U-trend plots and estimated ages that are stratigraphically consistent and in agreement with independent age estimates. We conclude that many marine terrace deposits are suitable for U-trend dating, but U-trend plots must be carefully evaluated and U-trend ages should be consistent with independent geologic control

Novices Outperform Experienced Laparoscopists on Virtual Reality Laparoscopy Simulator

BACKGROUND AND OBJECTIVES: Virtual reality has been poorly studied among gynecologic surgeons. The aim of this study was to evaluate whether performance on the Minimally Invasive Surgery Trainer-Virtual Reality (MIST-VR) laparoscopic trainer reflects laparoscopic experience among gynecologic surgeons and trainees. METHODS: Twenty-six medical students, residents, and attending gynecologic surgeons completed a MIST-VR training program. A new simulated task was then presented to each participant, who repeated the task until proficiency was reached. RESULTS: Attending physicians performed poorly when compared with medical students, requiring more than twice the number of attempts to reach proficiency (Mann-Whitney P\u3c0.01). Among medical students and residents, there was an association between years of live laparoscopy experience and poor simulator performance (Spearman r P=0.01). CONCLUSION: Increased operating room experience and age were associated with worsening simulator performance. Several potential explanations for this trend are discussed, including lack of tactile and contextual feedback. Caution should be exercised when considering current virtual reality simulator technology as a measure of experience or ability among gynecologic surgeons

Temporal dynamics in an immunological synapse:role of thermal fluctuations in signaling

The article analyzes the contribution of stochastic thermal fluctuations in the attachment times of the immature T-cell receptor TCR: peptide-major-histocompatibility-complex pMHC immunological synapse bond. The key question addressed here is the following: how does a synapse bond remain stabilized in the presence of high-frequency thermal noise that potentially equates to a strong detaching force? Focusing on the average time persistence of an immature synapse, we show that the high-frequency nodes accompanying large fluctuations are counterbalanced by low-frequency nodes that evolve over longer time periods, eventually leading to signaling of the immunological synapse bond primarily decided by nodes of the latter type. Our analysis shows that such a counterintuitive behavior could be easily explained from the fact that the survival probability distribution is governed by two distinct phases, corresponding to two separate time exponents, for the two different time regimes. The relatively shorter timescales correspond to the cohesion:adhesion induced immature bond formation whereas the larger time reciprocates the association:dissociation regime leading to TCR:pMHC signaling. From an estimate of the bond survival probability, we show that, at shorter timescales, this probability PΔ(τ) scales with time τ as a universal function of a rescaled noise amplitude DΔ2, such that PΔ(τ)∼τ-(ΔD+12),Δ being the distance from the mean intermembrane (T cell:Antigen Presenting Cell) separation distance. The crossover from this shorter to a longer time regime leads to a universality in the dynamics, at which point the survival probability shows a different power-law scaling compared to the one at shorter timescales. In biological terms, such a crossover indicates that the TCR:pMHC bond has a survival probability with a slower decay rate than the longer LFA-1:ICAM-1 bond justifying its stability

Contact time periods in immunological synapse

This paper resolves the long standing debate as to the proper time scale τ of the onset of the immunological synapse bond, the noncovalent chemical bond defining the immune pathways involving T cells and antigen presenting cells. Results from our model calculations show τ to be of the order of seconds instead of minutes. Close to the linearly stable regime, we show that in between the two critical spatial thresholds defined by the integrin:ligand pair (Δ2∼ 40-45 nm) and the T-cell receptor TCR:peptide-major-histocompatibility-complex pMHC bond (Δ1∼ 14-15 nm), τ grows monotonically with increasing coreceptor bond length separation δ (= Δ2-Δ1∼ 26-30 nm) while τ decays with Δ1 for fixed Δ2. The nonuniversal δ-dependent power-law structure of the probability density function further explains why only the TCR:pMHC bond is a likely candidate to form a stable synapse