Porcine-specific hemoglobin saturation measurements.

The determination of O(2) consumption by using arteriovenous O(2) content differences is dependent on accurate oxyhemoglobin saturation measurements. Because swine are a common experimental species, we describe the validation of CO-oximeter for porcine-specific oxyhemoglobin saturation. After developing a nonlinear mathematical model of the porcine oxyhemoglobin saturation curve, we made 366 porcine oxyhemoglobin saturation determinations with a calibrated blood-gas analyzer and a porcine-specific CO-oximeter. There was a high degree of correlation with minimal variability (r(2) = 0.99, SE of the estimate = 5.2%) between the mathematical model and the porcine-specific CO-oximeter measurements. Bland-Altman comparison showed that the CO-oximeter measurements were biased slightly lower (-0.4 vol%), and the limits of agreement (+/-2 SD) were 0.7 and -1.5 vol%. This is in contrast to a 10-20 vol% error if human-specific methods were used. The results show excellent agreement between the nonlinear model and CO-oximeter for porcine-specific oxyhemoglobin saturation measurements. In contrast, comparison of the porcine-specific oxyhemoglobin saturations with saturations obtained by using human methods highlights the necessity of species-specific measurement methodology.     

Identification, sequencing and mutagenesis of the gene for a d-carbamoylase from Agrobacterium radiobacter

Abstract A clone positive for d-carbamoylase activity (2.7 kb Hin dIII- Bam H1 DNA fragment) was obtained by screening a genomic library of Agrobacterium radiobacter in Escherichia coli . This DNA fragment contains an open reading frame of 912 bp which is predicted to encode a peptide of 304 amino acids with a calculated molecular mass of 34247 Da. The d-carbamoylase gene. named cauA , was placed under the control of T7 RNA-dependent promoter and expressed in E. coli BL21 (DE3). After induction with isopropyl-thio-β-d-galactopyranoside, the synthesis of d-carbamoylase in E. coli reached about 40% of the total protein. The expressed protein was shown to possess a molecular mass, on SDS-PAGE, of 36 kDa and showed an enhanced allowed us to establish that a Pro¹⁴→Leu¹⁴ exchange leads to an inactive enzyme species, while a Cys²⁷⁹→Ser²⁷⁹ exchange did not impair the functional properties of the enxyme.     

JAR human placental choriocarcinoma cells actively synthesize,take up and release polyamines

Uptake of polyamines has been investigated extensively in many cells, but not in placenta, where the polyamine– polyamine oxidase system is supposed to have an immunoregulatory function in pregnancy. Due to the importance of the transfer in this tissue, we have started this study. JAR human placental choriocarcinoma cells in monolayer at confluency were used as a model for measuring the key enzymes of polyamine synthesis and interconversion, rate of uptake and efflux, and the polyamine content. Polyamines were taken up by JAR cells and released by an independent mechanism. Ornithine decarboxylase and spermidine acetyltransferase activities and the rate of transport in and out of the cell were much higher than in other cells, such as L1210 cells. However the systems used for uptake and release appear in many respects to be similar to those observed in L1210 cells, but different from others. The uptake appears to be regulated by an inhibitory protein. Moreover, protein kinase C appears to be involved in the process. The efflux also is regulated as in L1210 cells, through control of H⁺ and Ca²⁺ concentration. In conclusion, this study shows that, in JAR cells, ornithine decarboxylase and spermidine acetyltransferase activities were much higher than in other cells, and so was the rate of transport in and out of the cells. As a result, a much higher polyamine content was observed.     

How the Context Matters. Literal and Figurative Meaning in the Embodied Language Paradigm

The involvement of the sensorimotor system in language understanding has been widely demonstrated. However, the role of context in these studies has only recently started to be addressed. Though words are bearers of a semantic potential, meaning is the product of a pragmatic process. It needs to be situated in a context to be disambiguated. The aim of this study was to test the hypothesis that embodied simulation occurring during linguistic processing is contextually modulated to the extent that the same sentence, depending on the context of utterance, leads to the activation of different effector-specific brain motor areas. In order to test this hypothesis, we asked subjects to give a motor response with the hand or the foot to the presentation of ambiguous idioms containing action-related words when these are preceded by context sentences. The results directly support our hypothesis only in relation to the comprehension of hand-related action sentences.     

Effects of Anabolic Steroids and High-Intensity Aerobic Exercise on Skeletal Muscle of Transgenic Mice

In an attempt to shorten recovery time and improve performance, strength and endurance athletes occasionally turn to the illicit use of anabolic-androgenic steroids (AAS). This study evaluated the effects of AAS treatment on the muscle mass and phenotypic characteristics of transgenic mice subjected to a high-intensity, aerobic training program (5d/wk for 6 weeks). The transgenic mice (CETP^+/-LDLr^-/+) were engineered to exhibit a lipid profile closer to humans. Animals were divided into groups of sedentary (Sed) and/or training (Ex) mice (each treated orally with AAS or gum arabic/vehicle: Sed-C, Sed-M, ex-C, ex-M). The effects of AAS (mesterolone: M) on specific phenotypic adaptations (muscle wet weight, cross-sectional area, and fiber type composition) in three hindlimb muscles (soleus:SOL, tibialis anterior:TA and gastrocnemius:GAS) were assessed. In order to detect subtle changes in fiber type profile, the entire range of fiber types (I, IC, IIAC, IIA, IIAD, IID, IIDB, IIB) was delineated using mATPase histochemistry. Body weight gain occurred throughout the study for all groups. However, the body weight gain was significantly minimized with exercise. This effect was blunted with mesterolone treatment. Both AAS treatment (Sed-M) and high-intensity, aerobic training (ex-C) increased the wet weights of all three muscles and induced differential hypertrophy of pure and hybrid fibers. Combination of AAS and training (ex-M) resulted in enhanced hypertrophy. In the SOL, mesterolone treatment (Sed-M and ex-M) caused dramatic increases in the percentages of fiber types IC, IIAC, IIAD, IID, with concomitant decrease in IIA, but had minimal impact on fiber type percentages in the predominantly fast muscles. Overall, the AAS-induced differential adaptive changes amounted to significant fiber type transformations in the fast-to-slow direction in SOL. AAS treatment had a significant effect on muscle weights and fiber type composition in SOL, TA and GAS which was even maximized in animals subjected to metabolically high-intensity aerobic exercise.     

Tetrahydro-β-carboline derivatives targeting fatty acid amide hydrolase (FAAH) and transient receptor potential (TRP) channels

A series of twenty-five derivatives of tetrahydro-β-carbolines 1–3 was synthesized and assayed on FAAH and TRPV1 and TRPA1 channels. Four carbamates, that is, 5a,c,e, and 9b inhibited FAAH with significant potency and interacted also effectively with TRPV1 and TRPA1 nociceptive receptors, while ureas 7b,d,f, and 8a,b were endowed with specific submicromolar TRPV1 modulating activities.     

Climatic change and its ecological implications at a subantarctic island

Summary Marion Island (47°S, 38°E) has one of the most oceanic climates on earth, with consistently low air temperatures, high precipitation, constantly high humidity, and low incident radiation. Since 1968 mean surface air temperature has increased significantly, by 0.025° C year^–1. This was strongly associated with corresponding changes in sea surface temperature but only weakly, or not at all, with variations in radiation and precipitation. We suggest that changing sealevel (atmospheric and oceanic) circulation patterns in the region underlie all of these changes. Sub-Antarctic terrestrial ecosystems are characterized by being species-poor and having a simple trophic structure. Marion Island is no exception and a scenario is presented of the implications of climatic change for the structure and functioning of its ecosystem. Primary production on the island is high and consequently the vegetation has a large annual requirement for nutrients. There are no macroherbivores and even the insects play only a small role as herbivores, so most of the energy and nutrients incorporated in primary production go through a detritus, rather than grazing, cycle. Ameliorating temperatures and increasing CO₂ levels are expected to increase productivity and nutrient demand even further. However, most of the plant communities occur on soils which have especially low available levels of nutrients and nutrient mineralization from organic reserves is the main bottleneck in nutrient cycling and primary production. Increasing temperatures will not significantly enhance microbially-mediated mineralization rates since soil microbiological processes on the island are strongly limited by waterlogging, rather than by temperature. The island supports large numbers of soil macro-arthropods, which are responsible for most of the nutrient release from peat and litter. The activities of these animals are strongly temperature dependent and increasing temperature will result in enhanced nutrient availability, allowing the potential for increased primary production due to elevated temperature and CO₂ levels to be realized. However, housemice occur on the island and have an important influence on the ecosystem, mainly by feeding on soil invertebrates. The mouse population is strongly temperature-limited and appears to be increasing, possibly as a result of ameliorating temperatures. We suggest that an increasing mouse population, through enhanced predation pressure on soil invertebrates, will decrease overall rates of nutrient cycling and cause imbalances between primary production and decomposition. This, along with more direct effects of mice (e.g. granivory) has important implications for vegetation succession and ecosystem structure and functioning on the island. Some of these are already apparent from comparisons with nearby Prince Edward Island where mice do not occur. Other implications of climatic change for the island are presented which emphasize the very marked influences that invasive organisms have on ecosystem structure and functioning. We suggest that changing sealevel circulation patterns, by allowing opportunities for colonization by new biota, may have an even more important influence on terrestrial sub-Antarctic ecosystems than is suggested merely on the basis of associated changes in temperature or precipitation.