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Recently, we characterized neoamphimedine (neo) as an ATP-competitive inhibitor of the ATPase domain of human Topoisomerase IIα. Thus far, neo is the only pyridoacridine with this mechanism of action. One limiting factor in the development of neo as a therapeutic agent has been access to sufficient amounts of material for biological testing. Although there are two reported syntheses of neo, both require 12 steps with low overall yields (≤6%). In this article, we report an improved total synthesis of neo achieved in 10 steps with a 25% overall yield. In addition, we report an expanded cytotoxicity study using a panel of human cancer cell lines, including: breast, colorectal, lung, and leukemia. Neo displays potent cytotoxicity (nM IC50 values) in all, with significant potency against colorectal cancer (lowest IC50 = 6 nM). We show that neo is cytotoxic not cytostatic, and that neo exerts cytotoxicity by inducing G2-M cell cycle arrest and apoptosis.

Using simulated falls performed under supervised conditions and activities of daily living (ADL) performed by elderly subjects, the ability to discriminate between falls and ADL was investigated using tri-axial accelerometer sensors, mounted on the trunk and thigh. Data analysis was performed using MATLAB to determine the peak accelerations recorded during eight different types of falls. These included; forward falls, backward falls and lateral falls left and right, performed with legs straight and flexed. Falls detection algorithms were devised using thresholding techniques. Falls could be distinguished from ADL for a total data set from 480 movements. This was accomplished using a single threshold determined by the fall-event data-set, applied to the resultant-magnitude acceleration signal from a tri-axial accelerometer located at the trunk.

The use of surface neuromuscular electrical stimulation (SNMES) in medicine is well established. However, discomfort has been identified as limiting the use of SNMES in these applications. This pilot study investigated the influence of various electrode sizes and their positioning on perceived pain and discomfort during neuromuscular electrical stimulation (NMES) of the gastrocnemius muscle using surface electrodes. This study formed part of a research project to develop a therapeutic device for calf muscle blood flow assist applications. Twelve healthy subjects (n=12) participated in this pilot study. Each participant attended the trial centre for testing which consisted of SNMES to four different electrode stimulation sites using two electrode sizes (round with areas 19.63 and 38.48 cm2). Comfort was assessed by asking the subjects to indicate the stimulation amplitude corresponding to the onset of discomfort (pain threshold) and the amplitude at which the discomfort became unbearable (pain tolerance). Of the four stimulation sites tested, two were deemed unsuccessful as it was very difficult to obtain a muscle contraction using these sites, while the remaining two sites elicited good muscle contraction. The most comfortable stimulation was achieved by placing the cathode electrode high on the calf, below the proximal end of the muscle heads and the anode electrode towards the end of the muscle belly and when the 19.63 cm2 electrodes were used at these sites (p=<0.001).

Progeny recovered from backcrossed transgenic maize tissue culture regenerants (R0) were analyzed to determine the segregation, expression, and stability of the introduced genes. Transgenic A188 x B73 R0 plants (regenerated from embryogenic suspension culture cells transformed by microprojectile bombardment; see [9]) were pollinated with nontransformed B73 pollen. Inheritance of a selectable marker gene, bar, and a nonselectable marker gene, uidA, was analyzed in progeny (R1) representing four independent transformation events. Activity of the bar gene product, phosphinothricin acetyltransferase (PAT), was assessed in plants comprising the four R1 populations. The number of R1 plants containing PAT activity per total number of R1 plants recovered for each population was 2/7, 19/34, 3/14 and 73/73. Molecular analysis confirmed the segregation of bar in three R1 populations and the lack of segregation in one R1 population. Cosegregation analysis indicated genetic linkage of bar and uidA in all four R1 populations. Analysis of numerous R2 plants derived from crossing transformed R1 plants with nontransformed inbreds revealed 1:1 segregation of PAT activity in three of four lines, including the line that failed to segregate in the R1 generation. Integrated copies of bar in one line appeared to be unstable or poorly transmitted.

A reproducible system for the generation of fertile, transgenic maize plants has been developed. Cells from embryogenic maize suspension cultures were transformed with the bacterial gene bar using microprojectile bombardment. Transformed calli were selected from the suspension cultures using the herbicide bialaphos. Integration of bar and activity of the enzyme phosphinothricin acetyltransferase (PAT) encoded by bar were confirmed in all bialaphos-resistant callus lines. Fertile transformed maize plants (R0) were regenerated, and of 53 progeny (R1) tested, 29 had PAT activity. All PAT-positive progeny analyzed contained bar. Localized application of herbicide to leaves of bar-transformed R0 and R1 plants resulted in no necrosis, confirming functional activity of PAT in the transgenic plants. Cotransformation experiments were performed using a mixture of two plasmids, one encoding PAT and one containing the nonselected gene encoding [beta]-glucuronidase. R0 plants regenerated from co-transformed callus expressed both genes. These results describe and confirm the development of a system for introduction of DNA into maize.

Erotomania is a mono-delusional disorder which, in its "pure" or "primary" form, should probably belong to the diagnostic category of Paranoia. Traditionally, the prognosis and response to treatment are very poor, but one of the present authors has previously proposed that the condition should respond to pimozide. Case histories of two patients successfully treated with this drug are presented.