Chemogenetic ON and OFF switches for RNA virus replication.

Therapeutic application of RNA viruses as oncolytic agents or gene vectors requires a tight control of virus activity if toxicity is a concern. Here we present a regulator switch for RNA viruses using a conditional protease approach, in which the function of at least one viral protein essential for transcription and replication is linked to autocatalytical, exogenous human immunodeficiency virus (HIV) protease activity. Virus activity can be en- or disabled by various HIV protease inhibitors. Incorporating the HIV protease dimer in the genome of vesicular stomatitis virus (VSV) into the open reading frame of either the P- or L-protein resulted in an ON switch. Here, virus activity depends on co-application of protease inhibitor in a dose-dependent manner. Conversely, an N-terminal VSV polymerase tag with the HIV protease dimer constitutes an OFF switch, as application of protease inhibitor stops virus activity. This technology may also be applicable to other potentially therapeutic RNA viruses.

Education and advice under ERISA for employee-directed 401(k) plans.

Section 404(c) regulation sets forth the conditions that plan fiduciaries must meet to be relieved of liability for the consequences of employees' control over their accounts. After reviewing applicable laws and regulations, the author concludes that employers desiring to provide employees with education and/or advice services through a third party should be able to do so while still obtaining reliance on the protections of Section 404(c) and without taking on significant additional fiduciary responsibilities.

FREEDOM TO OPERATE: Intellectual Property Protection in Plant Biology and Its Implications for the Conduct of Research.

▪ Abstract Research endeavors are being affected by issues involving intellectual property (patents, copyrights, and trademarks). The acquisition of rights in intellectual property by universities can result in the transfer of new innovations to the private sector, with the university recouping a share of the profits for support of further scientific research. Intellectual property rights available for new plant cultivars include plant patents, plant variety protection certificates, plant breeder's rights, and utility patents. Under the patent laws, there is no explicit exemption for research use, so researchers are increasingly being required to execute materials transfer agreements to obtain permission to use patented materials, such as techniques, genes, seeds, and cell lines, in laboratory research and in breeding programs. Research scientists must educate themselves on these issues so that they can make informed decisions regarding their research practices and the licensing of their discoveries.

Plant responses to environmental stress.

Considerable progress is being made in identifying genes that are important for tolerance to abiotic stress and in defining stress-responsive gene promoters and signal-transduction pathways. Although genetically engineered crop plants with greater resistance to environmental stress have not yet been produced, research is at a turning point where correlative changes can now be tested for effectiveness in conferring stress tolerance.

Regulation of the heat shock response in soybean seedlings.

The transcriptional response of soybean (Glycine max) seedlings during heat shock (HS) was investigated under two different treatment regimes. During prolonged heat treatment at 40 degrees C, active transcription of the HS genes (as measured by "runoff" transcription assays) occurs only during the first few hours. Nonetheless, mRNAs for these genes are present at relatively high abundance even after 9 hours of exposure to 40 degrees C. Because HS mRNAs have a fairly short half-life (less than 3 hours) at 28 degrees C, these results indicate that HS mRNAs are inherently more stable at 40 degrees C. During a second type of heat treatment regime-short pulses of high (45 degrees C) heat followed by 1 to 2 hours at 28 degrees C-transcription of HS genes is comparable to that achieved at 40 degrees C for the first few hours, even though the tissue is maintained at non-HS temperatures. The transcriptional responses to these two different heat treatments indicate that regulatory controls for the transcription of the HS genes must involve more than a simple sensing of ambient temperature, since transcription of these genes can be turned off at 40 degrees C (in the case of prolonged exposure) and can continue at 28 degrees C (following a short, severe heat treatment). Additional results demonstrate that the response of soybean seedlings to a particular HS depends on their prior exposure to heat; seedlings given a preheat treatment (that is known to induce thermotolerance) respond more moderately to a short heat pulse at 45 degrees C. Overall, this research indicates that plants have mechanisms for both monitoring the severity of changes in temperature and for measuring the magnitude and duration of the stress. Such information is then used to regulate the plant's response to heat both transcriptionally and posttranscriptionally.

Presence of Heat Shock mRNAs in Field Crown Soybeans.

Our laboratory has extensively defined many parameters of the heat shock (HS) response in etiolated soybean (Glycine max [L.] Merr.) hypocotyls, including the identification of cDNA clones for mRNAs encoding several low molecular weight HS proteins. We have now investigated the response of mature plants to a HS in a growth chamber and to high temperature stress under field conditions. Soybean plants show induction of HS mRNAs when the temperature of the chamber is rapidly shifted from 28 degrees C to 45 degrees C. This temperature of induction is significantly higher than the optimal induction temperature for etiolated hypocotyls, probably reflecting the ability of mature plants to lower their leaf temperatures below the ambient air temperature through transpirational cooling. Samples of soybean leaves were taken from an irrigated and a nonirrigated field during a 24-h period when midday temperatures reached 40 degrees C. Several HS mRNAs were present in samples from both fields, although the levels of these mRNAs were much higher in nonirrigated leaves. This differential response of HS mRNA steady state levels was not a response to water stress, since water-stressed plants at 28 degrees C did not induce HS mRNAS. Rather, these quantitative differences are probably due to differences in actual leaf temperatures between irrigated and nonirrigated leaves. The presence of these HS mRNAS in field-grown plants suggests that HS proteins are produced as part of the normal plant response to high temperature.

Metabolic Regulation during Glyceollin Biosynthesis in Green Soybean Hypocotyls.

The accumulation of the isoflavonoid phytoalexin, glyceollin, occurs in hypocotyls of green soybean seedlings (Glycine max L. Merr. cv Harosoy 63) in response to the injection of a glucan elicitor isolated from the mycelial walls of the fungus, Phytophthora megasperma f. sp. glycinea. This accumulation, which levels off after 24 hours, is preceded by a dramatic, transient rise in extractable activities of two early enzymes in the biosynthetic pathway, phenylalanine ammonia-lyase (PAL) and p-coumaryl CoA ligase (pCL). The maximum amount of extractable activity occurs 12 to 16 hours after elicitor treatment and is coincident with the most rapid period of glyceollin accumulation. These results suggest a regulatory role for these early enzymes in the biosynthesis of this secondary metabolite. High performance liquid chromatography analysis of the early intermediates in the pathway further corroborates this hypothesis. The relative pool size and rate of turnover of p-coumaric acid, an early intermediate in glyceollin production, increase during the period of rapid increases in enzyme activities. Removal of cotyledons from elicitor-treated seedlings reduces glyceollin accumulation approximately 70%. This limitation of phytoalexin accumulation by cotyledon removal is correlated with a similar cotyledon effect on reduction of extractable activities of both PAL and pCL as well as a decrease in the flux of carbon through the p-coumaric acid pool. This research further supports the hypothesis that early enzymic steps in a biosynthetic pathway diverting carbon from primary to secondary metabolites function as regulatory control points.

[Oligodendroglioma. Clinical course in relation to histological grading]. / Das Oligodendrogliom. Der klinische Verlauf in bezug zum histologischen Grading.

The history of 99 patients, operated on for oligodendroglioma, has been related to the microscopic picture of the tumours which were graded in three stages of malignancy. Clinical course and histological grading fit together well. Certain signs and symptoms correlate with the kind of tumour. The prognosis in oligodendroglioma is as bad as in astrocytoma. The danger of recurrence is high, even after radical operation. Radiation therapy is useless.

Activation of endogenous respiration and anion transport in corn mitochondria by acidification of the medium.

Acidification of the suspending medium of corn mitochondria (Zea mays L., WF9 x Mo17) from pH 7.5 to pH 6.8 to 6.4 initiates osmotic swelling with the transportable anions citrate, sulfate, and phosphate. Swelling becomes pronounced with a combination of citrate plus sulfate or phosphate. Acidification proves to activate endogenous respiration, which is essentially zero at pH 7.5. The endogenous respiration transports citrate (in the presence of sulfate or phosphate) which then contributes to respiration and the accelerated osmotic swelling. Mersalyl will inhibit the swelling and antimycin inhibits the endogenous respiration. Magnesium appears to reduce the permeability of the membranes under the acid conditions.