Repression of SV40 T oncoprotein expression by DMSO.

SV40 large T oncoprotein-transformed murine mesenchymal 3T3 T stem cells (CSV3 cells) can be induced to growth arrest and then differentiate into adipocytes. When differentiation occurs, SV40 T oncoprotein expression is repressed (Estervig et al., J Virol 63:2718, 1989). To determine if repression of T oncoprotein expression can also be induced pharmacologically, the effect of a variety of agents that have been reported to effect differentiation in various cell types but not in 3T3 T or CSV3 cells was tested. This rationale suggests that if any of these agents repress T oncoprotein expression in CSV3 cells, then the results would establish that repression of T oncoprotein expression can be mediated by mechanisms independent of overt differentiation. The results show that dimethylsulfoxide (DMSO) is the only agent tested that represses T oncoprotein expression in CSV3 cells. Repression occurs in a dosage-dependent manner within 24-96 hours after exposure to DMSO. The effect of DMSO on T oncoprotein expression is mediated by posttranslational mechanisms that decrease the stability of the T oncoprotein. DMSO-induced repression of T oncoprotein expression is also associated with reversion of the transformed phenotype in CSV3 cells as demonstrated by the loss of responsiveness to a specific transformation-associated mitogen. These data support the conclusion that the pharmacological repression of T oncoprotein expression represents a form of cancer suppressor activity that can be mediated by a distinct molecular mechanism.     

Statistical Comparison of Band Spectral Powers: An Aid in Stochastical Analysis of Brain Electrical Activity. Part 2: Construction of Confidence Intervals of Spectral Band Power and their Application in EEG-Analysis

Based on the result that the distribution of estimated spectral power in broad frequency bands can be approximated by x²-distribution the accuracy of the approximation was tested by different models of stochastical processes. Two significance tests for comparison of spectral band powers on the basis of this approximation and the estimation of confidence intervals were developed and tested by application on EEG data. By means of these test statistics, a new basis for statistical comparison between EEG maps of groups as well as between maps of groups and single maps can be suggested.     

Scale dependency and fractal dimension of rarity

The rarity of species in a country is usually determined by counting the number of grid cells occupied by those species on a geographical observation grid. In this paper, we present a measure of rarity that is less sensitive to the shape and size of a country. We demonstrate that the distribution of species on a national grid is not monofractal. Consequently, rarity figures cannot be scaled down to a finer grid merely using scale-area plots. We propose a downscaling method that takes into account the non-monofractal distribution of species. Rarity figures have often been published on a scale comprising a limited number of rarity classes. This article finally provides an insight into the degree of accuracy of such classes.     

Quantitative analysis of trifluoroacetic acid in body fluids of patients treated with halothane

A simple procedure for the quantitative analysis of trifluoroscetic acid (TFA) in urine and serum from patients narcotized with halothane is described. This involves addition of sodium hydroxide to the body fluid, evaporation of the aqueous phase and esterification of TFA in concentrated sulphuric acid with 2,2,2-trichloroethanol. The gaseous phases above the reaction mixture were then analyzed by gas chromatography with a nickel-63 electron-capture detector. The detection limit was 1 μg of TFA per mililitre of body fluid (200 μg of body fluid are analysed) and the relative standard deviation was ±6%. Patients treated with ethrane, another commercial ansesthetic, did not produce any detectable TFA.     

High resolution ultrasound-guided microinjection for interventional studies of early embryonic and placental development in vivo in mice

Background  

In utero microinjection has proven valuable for exploring the developmental consequences of altering gene expression, and for studying cell lineage or migration during the latter half of embryonic mouse development (from embryonic day 9.5 of gestation (E9.5)). In the current study, we use ultrasound guidance to accurately target microinjections in the conceptus at E6.5–E7.5, which is prior to cardiovascular or placental dependence. This method may be useful for determining the developmental effects of targeted genetic or cellular interventions at critical stages of placentation, gastrulation, axis formation, and neural tube closure.     

Ceftiofur derivates in serum and endometrial tissue after intramuscular administration in healthy mares

Endometritis is one of the major problems in the horse breeding industry. The use of antibiotics for treatment of endometritis in the mare is recommended as best practice. The intrauterine application of antibiotics, however, has been under discussion over the last years because of concerns about its efficacy. The systemic use of antibiotics has been considered more effective because of its better distribution within the uterus. The objective of the present study was to determine the concentration of ceftiofur derivates in serum and endometrial tissue after intramuscular administration. Specifically, the authors tested the hypothesis that ceftiofur concentrations in serum and endometrial tissue remain above the minimum inhibitory concentration (MIC) for common uterine pathogens for 24 h. Nine mares in estrus received a single dose of 2.2 mg/kg ceftiofur hydrochloride intramuscular per kg of body weight. Blood samples and endometrial tissue were obtained immediately before treatment (−1 h) and 2 h and 24 h after treatment. Endometrial tissue was collected with a Kevorkian biopsy punch. Additional blood samples were collected 4 h and 10 h after treatment from the jugular veins. For determination of ceftiofur derivates in serum and endometrial tissue a high performance liquid chromatography (HPLC) assay was used. Results in serum and uterine tissue revealed greatest concentration of ceftiofur at 2 h and lowest concentrations at 24 h after treatment. Concentrations of ceftiofur at 2 and 24 h after treatment were significantly greater in serum than in endometrial tissue, but remained above the reported MIC for Streptococcus equi zooepidemicus and Escherichia coli in both serum and endometrial tissue until 24 h after treatment.     

Circadian Changes in the Surface Area of Renal Glomeruli from Normal Rats

In an attempt to further investigate circadian changes in kidney function, the planar surface area (PSA) (µm 2) of renal isolated glomeruli from normal rats was monitored using a computerized image analyzer method. Eight male Sprague-Dawley (SPRD) rats, aged 12û14 weeks, were entrained to a 12-h light/12-h dark cycle in controlled environmental conditions of temperature (22±2°C) two weeks before the experiments started. Isolated glomerular preparations were obtained, using a mechanical sieving technique, at four different circadian times: 07:00, 13:00, 19:00 and 01:00. It was the finding of the present study that the PSA of the glomeruli varied significantly over the 24-h period, but showed a weak amplitude. The size of the glomeruli reached the highest values at night (21358.59±456.72 µm 2 at 21:17) with an acrophase at 21:39, as do all the other renal parameters, but also blood pressure and many vasoactive compounds involved in the regulation of the mesangial cell physiology, contractile element of the glomerulus. Such chronobiological data not only provided a clear example of complementarity between in vivo and ex vivo experiments, but evidenced that temporal changes do exist in kidney structure and could be correlated with rhythms in renal physiology.     

Retrovirus purification: method that conserves envelope glycoprotein and maximizes infectivity.

A Sepharose 4B chromatographic method for purification of retroviruses is described which was less time consuming, increased purified virus yields, conserved viral glycoprotein, and increased recovery of biological infectivity in comparison with conventional sucrose gradient ultracentrifugation techniques.     

Beyond reduction of atherosclerosis: PON2 provides apoptosis resistance and stabilizes tumor cells

Major contributors to atherosclerosis are oxidative damage and endoplasmic reticulum (ER) stress-induced apoptosis; both of which can be diminished by the anti-oxidative protein paraoxonase-2 (PON2). ER stress is also relevant to cancer and associated with anti-cancer treatment resistance. Hence, we addressed, for the first time, whether PON2 contributes to tumorigenesis and apoptotic escape. Intriguingly, we found that several human tumors upregulated PON2 and such overexpression provided resistance to different chemotherapeutics (imatinib, doxorubicine, staurosporine, or actinomycin) in cell culture models. This was reversed after PON2 knock-down. Remarkably, just deficiency of PON2 caused apoptosis of selective tumor cells per se, demonstrating a previously unanticipated oncogenic function. We found a dual mechanistic role. During ER stress, high PON2 levels lowered redox-triggered induction of pro-apoptotic CHOP particularly via the JNK pathway, which prevented mitochondrial cell death signaling. Apart from CHOP, PON2 also diminished intrinsic apoptosis as it prevented mitochondrial superoxide formation, cardiolipin peroxidation, cytochrome c release, and caspase activation. Ligand-stimulated apoptosis by TRAIL or TNFα remained unchanged. Finally, PON2 knock-down caused vast reactive oxygen species formation and stimulated JNK-triggered CHOP expression, but inhibition of JNK signaling did not prevent cell death, demonstrating the pleiotropic, dominating anti-oxidative effect of PON2. Therefore, targeting redox balance is powerful to induce selective tumor cell death and proposes PON2 as new putative anti-tumor candidate.