Molecular Electroporation and the Transduction of Oligoarginines

Certain short polycations, such as TAT and polyarginine, rapidly pass through the plasma membranes of mammalian cells by an unknown mechanism called transduction as well as by endocytosis and macropinocytosis. These cell-penetrating peptides (CPPs) promise to be medically useful when fused to biologically active peptides. I offer a simple model in which one or more CPPs and the phosphatidylserines of the inner leaflet form a kind of capacitor with a voltage in excess of 180 mV, high enough to create a molecular electropore. The model is consistent with an empirical upper limit on the cargo peptide of 40--60 amino acids and with experimental data on how the transduction of a polyarginine-fluorophore into mouse C2C12 myoblasts depends on the number of arginines in the CPP and on the CPP concentration. The model makes three testable predictions.Comment: 15 pages, 5 figure

Identification of microtubule-associated proteins in the centrosome, spindle, and kinetochore of the early Drosophila embryo.

We have developed affinity chromatography methods for the isolation of microtubule-associated proteins (MAPs) from soluble cytoplasmic extracts and have used them to analyze the cytoskeleton of the early Drosophila embryo. More than 50 Drosophila embryo proteins bind to microtubule affinity columns. To begin to characterize these proteins, we have generated individual mouse polyclonal antibodies that specifically recognize 24 of them. As judged by immunofluorescence, some of the antigens localize to the mitotic spindle in the early Drosophila embryo, while others are present in centrosomes, kinetochores, subsets of microtubules, or a combination of these structures. Since 20 of the 24 antibodies stain microtubule structures, it is likely that most of the proteins that bind to our columns are associated with microtubules in vivo. Very few MAPS seem to be identically localized in the cell, indicating that the microtubule cytoskeleton is remarkably complex

Microwave power coupling in a surface wave excited plasma

In recent decades, different types of plasma sources have been used for various types of plasma processing, such as, etching and thin film deposition. The critical parameter for effective plasma processing is high plasma density. One type of high density plasma source is Microwave sheath-Voltage combination Plasma (MVP). In the present investigation, a better design of MVP source is reported, in which over-dense plasma is generated for low input microwave powers. The results indicate that the length of plasma column increases significantly with increase in input microwave power.Comment: 10 pages, 5 figure

The Intermediate Disorder Regime for Directed Polymers in Dimension 1+1

We introduce a new disorder regime for directed polymers with one space and one time dimension that is accessed by scaling the inverse temperature parameter \beta with the length of the polymer n. We scale \beta_n := \beta n^{-\alpha} for alpha non-negative. This scaling sits in between the usual weak disorder (\beta = 0) and strong disorder regimes (\beta > 0). The fluctuation exponents zeta for the polymer endpoint and \chi for the free energy depend on \alpha in this regime, with \alpha = 0 corresponding to the usual polymer exponents \zeta = 2/3, \chi = 1/3 and \alpha >= 1/4 corresponding to the simple random walk exponents \zeta = 1/2, \chi = 0. For 0 < \alpha < 1/4 the exponents interpolate linearly between these two extremes. At \alpha = 1/4 we exactly identify the limiting distribution of the free energy and the end point of the polymer.Comment: 4 pages, 1 figure. Added a more detailed description of scaling results in the critical regime. Improved simulation results

Studies on the mechanism of retinoid-induced pattern duplications in the early chick limb bud: temporal and spatial aspects.

All-trans-retinoic acid causes striking digit pattern changes when it is continuously released from a bead implanted in the anterior margin of an early chick wing bud. In addition to the normal set of digits (234), extra digits form in a mirror-symmetrical arrangement, creating digit patterns such as a 432234. These retinoic acid-induced pattern duplications closely mimic those found after grafts of polarizing region cells to the same positions with regard to dose-response, timing, and positional effects. To elucidate the mechanism by which retinoic acid induces these pattern duplications, we have studied the temporal and spatial distribution of all-trans-retinoic acid and its potent analogue TTNPB in these limb buds. We find that the induction process is biphasic: there is an 8-h lag phase followed by a 6-h duplication phase, during which additional digits are irreversibly specified in the sequence digit 2, digit 3, digit 4. On average, formation of each digit seems to require between 1 and 2 h. The tissue concentrations, metabolic pattern, and spatial distribution of all-trans-retinoic acid and TTNPB in the limb rapidly reach a steady state, in which the continuous release of the retinoid is balanced by loss from metabolism and blood circulation. Pulse-chase experiments reveal that the half-time of clearance from the bud is 20 min for all-trans-retinoic acid and 80 min for TTNPB. Manipulations that change the experimentally induced steep concentration gradient of TTNPB suggest that a graded distribution of retinoid concentrations across the limb is required during the duplication phase to induce changes in the digit pattern. The extensive similarities between results obtained with retinoids and with polarizing region grafts raise the possibility that retinoic acid serves as a natural "morphogen" in the limb