In vivo electrochemical studies of dopamine clearance in subregions of rat nucleus accumbens: differential properties of the core and shell

The effects of the rodent hepatocarcinogens clofibric acid and diprofibrate on the activity of the peroxisomal fatty acyl-CoA oxidase, DNA synthesis, and apoptosis were compared in cultured rat and human hepatocytes. Rat hepatocytes expressed a 10-fold greater level of the peroxisomal fatty acyl-CoA oxidase compared to human hepatocytes. At the highest concentration (1.0 mM), both drugs induced a two- to threefold increase in this enzyme activity in both rat and human hepatocytes. Ciprofibrate (0.1 and 0.2 mM) caused a twofold increase in DNA synthesis in rat hepatocytes, whereas clofibric acid had no effect on DNA synthesis in these cells. In contrast, increasing concentrations of both clofibric acid and ciprofibrate produced inhibition of DNA synthesis in human hepatocytes. By using the terminal transferase dUTP-biotin nick end labeling technique, it was observed that 0.1 and 0.2 mM clofibric acid and ciprofibrate suppressed transforming growth factor-beta (TGF beta)-induced apoptosis by 50% in rat hepatocytes, but they had no effect on TGF beta-induced apoptosis in human hepatocytes. Although clofibric acid and ciprofibrate diminished TGF beta-induced apoptosis, they had no effect on the basal apoptotic levels in the rat hepatocyte cultures. However, both drugs significantly increased the percent of apoptotic cells in the human hepatocyte cultures. It is concluded that primary rat and human hepatocyte cultures respond differently to peroxisome proliferators. The differences in effects on DNA synthesis and apoptosis support the hypothesis that human liver cells are refractory to peroxisome proliferator-induced hepatocarcinogenesis.     

A Golgi localization signal identified in the Menkes recombinant protein

Menkes disease arises from a genetic impairment in copper transport. The gene responsible for the phenotype has been identified as a copper transporting ATPase ( ATP7A ). Recently, the protein encoded by the ATP7A gene has been localized to the Golgi complex. In order to investigate the role of the Menkes disease protein in copper transport, recombinant constructs containing both the full-length open reading frame and an alternatively spliced form have been successfully expressed and localized in mammalian cells. Other studies of a patient with occipital horn syndrome, an allelic variant of Menkes disease, have demonstrated that only this alternatively spliced isoform and not the full-length form is expressed in this patient. The milder form of this patient's phenotype suggests that the alternatively spliced isoform has some functional role in copper transport. In the present study the full-length recombinant Menkes protein was shown by immunofluorescence to localize to the Golgi apparatus and the alternatively spliced form, lacking sequences for transmembrane domains 3 and 4 encoded by exon 10, was shown to localize to the endoplasmic reticulum. Using sequences from exon 10 fused to a non-Golgi reporter molecule, a 38 amino acid sequence containing transmembrane domain 3 of the Menkes protein was found to be sufficient for localization to the Golgi complex. Therefore, the protein sequence encoded by exon 10 may be responsible for this differential localization and both isoforms may be required for comprehensive transport of copper within the cell.     

Inhibition of ingestive behavior following fourth ventricle bombesin injection in chronic decerebrate rats.

Tested the hypothesis that the effects of 4th ventricle bombesin (BN) injection on feeding require interaction with forebrain neural systems by measuring intraoral sucrose (0.1 M) in tube-fed control and tube-fed supracollicular decerebrate rats after 4th ventricle injections of 1, 5, 10, 20, and 50 ng BN. Fourth ventricle injections of all doses of BN reliably suppressed sucrose intake in both control and chronic decerebrate rats. These results indicate that caudal brain-stem afferent signals produced by 4th ventricle BN injections are integrated by the local neural circuitry of the caudal brain stem, independent of the forebrain systems, to modulate ingestive behavior. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Nucleic acid delivery: the missing pieces of the puzzle?

The delivery of genes or RNA interference (RNAi) agents can increase or decrease the expression of virtually any protein in a cell, and this process opens the path for cures to most diseases that afflict humans. However, the high molecular weight, anionic nature, and instability of nucleic acids in the presence of enzymes pose major obstacles to their delivery and frustrates their use as human therapies. This Account describes current ideas about the mechanisms in nonviral nucleic acid delivery and how lipidic and polymeric carriers can overcome some of the critical barriers to delivery. Over the last 20 years, researchers have developed a multitude of polymeric and lipidic vectors, but only a small fraction of these have progressed into clinical trials. None of these vectors has received FDA approval, which indicates that the current vectors do not yet have suitable properties for effective in vivo nucleic acid delivery. Nucleic acid delivery is a multistep process and inefficiencies at any stage result in a dramatic decrease in gene delivery or gene silencing. However, the majority of studies investigating synthetic vectors focus solely on optimization of endosomal escape. A small number of studies address how to improve uptake via targeted delivery, and an even smaller fraction examine the intracellular fate of the delivery systems and nucleic acid cargo. The internalization of genes into the cell nucleus remains an inefficient and mysterious process. In the case of DNA delivery, strategies are needed to increase and accelerate the migration of DNA through the cytoplasm and transport it through the nuclear membrane. siRNA delivery involves fewer barriers. siRNA is more readily released from the carrier and more resistant to enzymatic degradation, and its target is in the cytoplasm; hence, siRNA delivery systems are becoming a clinical reality. With regard to siRNA therapy, the exact cytoplasmic location of RNA-induced silencing complex (RISC) formation and activity is unknown, which makes specific targeting of the RISC for more efficient delivery difficult. Furthermore, we would like to identify the factors that favor the binding of siRNA to Ago-2. If we could understand how the half-life of siRNA and Ago-2/siRNA complex in the cytoplasm can be modulated without interfering with RISC functions that are essential for normal cell activity, we could increase siRNA delivery efficiency. In this Account, we review the current synthetic vectors and propose alternative strategies in a few cases. We also suggest how certain cellular mechanisms might be exploited to improve gene transfection and silencing. Finally, we discuss whether some carriers that deliver the siRNA to cells could also repackage the siRNA into exosomes. The exosomes would then transport the siRNA into a subsequent population of cells that manifest the siRNA effect. This piggy-back mechanism may be responsible for reported deep tissue siRNA effects using certain carriers.     

Smoke gas analysis by Fourier transform infrared spectroscopy – summary of the SAFIR project results

The determination of toxic components from fire gases is difficult because the environment is hot, reactions are often temperature dependent, and a lot of soot may be produced. Due to the different properties of the gas components, a different time‐consuming procedure for each species has traditionally been used. The use of FTIR (Fourier transform infrared) spectrometers as a continuous monitoring technique overcomes many of the problems in smoke gas analyses. FTIR offers an opportunity to set up a calibration and prediction method for each gas showing a characteristic spectral band in the infrared region of the spectrum. The objective of the SAFIR project was to further develop the FTIR gas analysis of smoke gases to be an applicable and reliable method for the determination of toxic components in combustion gases related to fire test conditions. The optimum probe design, filter parameters and the most suitable sampling lines in terms of flow rate, diameter, construction material and operating temperature have been specified. In the large scale, special concern was given to the probe design and the effects of the probe location as well as practical considerations of the sampling line length. Quantitative calibration and prediction methods have been constructed for different components present in smoke gases. Recommendations on how to deal with interferents, non‐linearities and outliers have been provided and a verification method for the spectrometer for unexpected variations and for the different models have been described. FTIR measurement procedures in different fire test scenarios have been studied using the recommendations of this project for measurement techniques and analysis and an interlaboratory trial of the FTIR technique in smoke gas analysis was carried out to define the repeatability and reproducibility of the method in connection with a small scale fire test method, the cone calorimeter. Copyright © 2000 John Wiley & Sons Ltd.     

Fluorescence characteristics of thermo-altered exinites (sporinites)

Fluorescence spectra of heat-treated exinites (sporinites) vary systematically with increasing temperatures of treatment. The wavelength of the fluorescence maximum, λ_max, shifts toward the longer wavelength as a result of thermal alteration. The maxima shift from 560 nm in untreated lignite to 640 nm in lignite after heating to 350 °C. Numerical values, such as fluorescence maximum, λ_max, and spectral quotient, $\text{Q}{}_{\mathrm{S}}\text{=}\text{I}{}_{650}{}_{\mathrm{I}}\text{500}$, can be used as rank parameters of coals. The technique can also be used as a tool to monitor thermal reactions of coals at relatively moderate temperatures. The method can distinguish certain resinites from exinites.     

Olfactory Responses to Aphid and Host Plant Volatile Releases: (E)-β-Farnesene an Effective Kairomone for the Predator Adalia bipunctata

The volatiles released from several aphid and host plant species, alone or associated, were studied for their infochemical role in prey location. Using a four-arm olfactometer, the attraction of several combinations of three aphid (Myzus persicae, Acyrthosiphon pisum, and Brevicoryne brassicae) and three plant (Vicia faba, Brassica napus, and Sinapis alba) species toward Adalia bipunctata larvae and adults was observed. Both predatory larvae and adults were attracted only by A. pisum and M. persicae when they were crushed, whatever the host plant. (E)-beta-farnesene, the aphid alarm pheromone, was the effective kairomone for the ladybird. Plant leaves alone (V. faba, B. napus, and S. alba) or in association with nonstressed whole aphids (the three species) did not have any attraction for the predator. The B. brassicae specialist aphid is the only prey that was not attracted to A. bipunctata larvae and adults, even if they were crushed. Release of B. brassicae molecules similar to the host plant allelochemicals was demonstrated by GC-MS analysis. The lack of behavioral response of the ladybird at short distance toward the cruciferous specialist aphid was related only to the absence of (E)-beta-farnesene in the aphid prey volatile pattern.     

Thermoplastic olefin/clay nanocomposites: Morphology and mechanical properties

Thermoplastic olefin (TPO)/clay nanocomposites were made with clay loadings of 0.6–6.7 wt %. The morphology of these TPO/clay nanocomposites was investigated with atomic force microscopy, transmission electron microscopy (TEM), and X‐ray diffraction. The ethylene–propylene rubber (EPR) particle morphology in the TPO underwent progressive particle breakup and decreased in particle size as the clay loading increased from 0.6 to 5.6 wt %. TEM micrographs showed that the clay platelets preferentially segregated to the rubber–particle interface. The breakup of the EPR particles was suspected to be due to the increasing melt viscosity observed as the clay loading increased or to the accompanying chemical modifiers of the clay, acting as interfacial agents and reducing the interfacial tension with a concomitant reduction in the particle size. The flexural modulus of the injection moldings increased monotonically as the clay loading increased. The unnotched (Izod) impact strength was substantially increased or maintained, whereas the notched (Izod) impact strength decreased modestly as the clay loading increased. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 928–936, 2004     

Field assisted and flash sintering of alumina and its relationship to conductivity and MgO-doping

We show that flash-sintering in MgO-doped alumina is accompanied by a sharp increase in electrical conductivity. Experiments that measure conductivity in fully dense specimens, prepared by conventional sintering, prove that this is not a cause-and-effect relationship, but instead that the concomitant increase in the sintering rate and the conductivity share a common mechanism. The underlying mechanism, however, is mystifying since electrical conductivity is controlled by the transport of the fastest moving charged species, while sintering, which requires molecular transport or chemical diffusion, is limited by the slow moving charged species. Joule heating of the specimen during flash sintering cannot account for the anomalously high sintering rates. The sintering behavior of MgO-doped alumina is compared to that of nominally pure-alumina: the differences provide insight into the underlying mechanism for flash-sintering. We show that the pre-exponential in the Arrhenius equation for conductivity is enhanced in the non-linear regime, while the activation energy remains unchanged. The nucleation of Frenkel pairs is proposed as a mechanism to explain the coupling between flash-sintering and the non-linear increase in the conductivity.