Determination of new 2,3-benzodiazepines in rat plasma using high-performance liquid chromatography with ultraviolet detection

A method for the analysis of [1-(4-aminophenyl)-3,5-dihydro-7,8-dimethoxy-4H-2,3-benzodiazepin-4-one] (CFM-2) and its analogues CFM-3, CFM-4 and CFM-5 in rat plasma was developed. The 2,3-benzodiazepines (2,3-BZs) were extracted by liquid–liquid extraction and analyzed using high-performance liquid chromatography (HPLC) with ultraviolet detection (UV) at 240 nm. The method exhibited a large linear range from 0.05 to 2 μg/ml with an intra-assay accuracy for all studied compounds ranging from 92 to 105.5%; whereas the intra-assay precision ranged from 0.59 to 8.16% in rat plasma. The inter-assay accuracy of CFM-2, CFM-4 and their 3-methyl derivatives, CFM-3 and CFM-5 ranged from 92.2 to 107% and the inter-assay precision ranged from 2.17 to 11.9% in rat plasma. The lower limit of detection was 5.5 ng/ml for CFM-2, 6.5 ng/ml for CFM-3, 7 ng/ml for CFM-4 and 8.5 ng/ml for CFM-5 in rat plasma. The pharmacokinetic study demonstrated that 2,3-BZs achieved a peak plasma concentration between 45 and 75 min after drug administration. Moreover, we observed that plasma chromatograms of rats treated with CFM-3, CFM-4 and CFM-5, respectively, showed a peak consistent with CFM-2. Our study suggests that CFM-4, CFM-5 and CFM-3 are prodrugs of CFM-2, in which they are biotransformed in vivo via different metabolic pathways. In particular, CFM-2 has been proven to possess anticonvulsant activity in various models of seizures, acting as α-amino-3-hydroxy-5-methyl-isoxazole-4-propionate (AMPA) receptor antagonist.     

Development of 5-hydroxypyrazole derivatives as reversible inhibitors of lysine specific demethylase 1

A series of reversible inhibitors of lysine specific demethylase 1 (LSD1) with a 5-hydroxypyrazole scaffold have been developed from compound 7, which was identified from the patent literature. Surface plasmon resonance (SPR) and biochemical analysis showed it to be a reversible LSD1 inhibitor with an IC₅₀ value of 0.23 µM. Optimisation of this compound by rational design afforded compounds with K_d values of <10 nM. In human THP-1 cells, these compounds were found to upregulate the expression of the surrogate cellular biomarker CD86. Compound 11p was found to have moderate oral bioavailability in mice suggesting its potential for use as an in vivo tool compound.     

Drought stress memory in the photosynthetic mechanisms of an invasive CAM species, <Emphasis Type="Italic">Aptenia cordifolia</Emphasis>

Plants are known for their high capacity to acclimatise to fluctuating environmental conditions. A wide range of environmental conditions can lead to suboptimal physiological efficiency. However, recent studies have shown that plants can withstand repeated periods of stress. To find out how they do it, we studied photosynthetic adjustments to repeated water stress in Aptenia cordifolia: a facultative, invasive CAM species. Plants were subjected to three cycles of water deficit, and photosynthetic parameters and chloroplast antioxidants were quantified to gain an understanding of the mechanisms by which they cope with repeated stress periods. Significant modification of the photosystems’ antenna and reaction centres was observed in plants subjected to previous water stress cycles, and this led to higher PSII efficiency than in plants challenged with drought for the first time. These findings underline the biological significance of stress memory and show how plants can adjust their photosynthetic apparatus to fluctuating environmental conditions and thus optimise photosynthesis and photoprotection under drought conditions.     

Impact of the non-indigenous shrub species Spartium junceum (Fabaceae) on native vegetation in central Spain

Aims The introduction of potentially invasive species through ornamental cultivation or for rehabilitation purposes is a serious environmental problem. They cause damage to biodiversity through loss, increased mortality or ' in situ ' selection phenomena in natural flora. Spartium junceum is a Mediterranean shrub that is not native in most areas of the Iberian Peninsula, although it is extensively grown for the rehabilitation of roadsides. We have investigated the effect on the native vegetation of an old S. junceum (Fabaceae) plantation in a conservation area in the centre of the Iberian Peninsula: the Cuenca Alta del Manzanares nature reserve in Madrid.Methods We compared S. junceum stands with the native nanophanerophytic Cistus ladanifer community at different ecosystem properties: soil properties, temporal soil seed bank contents, standing vegetation and net primary production of annual grasslands growing in these shrublands.Important findings The results highlighted S. junceum 's ability to become established in the new environment (marginal areas of the nature reserve) and ensure its successful growth. This is more apparent in northern and eastern exposures where this formation contacts with the core of the best conserved native vegetation in the nature reserve. Soils under Spartium showed a higher nitrogen content, indicating its capacity—in common with other legumes—to fix N, and conferring an advantage over Cistus, which is N-limited. Other soil nutrients such as phosphorus, magnesium and calcium and water availability are also higher in Spartium soils than in Cistus. Phosphorus is usually a constraint for N-fixers, but in our study, it is the most significant soil variable in both shrub formations and is important to the success of Spartium. Water availability is a key factor for Mediterranean vegetation, and particularly in autumn when soils are recharged. The Spartium formation is able to retain water as its growth produces a closer canopy than Cistus, thereby preventing water evaporation and contributing to the success of this species. Perennials are more frequent in the standing vegetation than in the seed bank, whereas therophytes are similar. The standing vegetation has therophytes and chamaephytes as the predominant growth forms in Spartium sites, and hemicryptophytes and phanerophytes in Cistus. Therophytes are dominant in Spartium and Cistus seed banks, although the first formation has more species. Spartium has a higher number of hemicryptophytes and Cistus is higher in phanerophytes. Northern and eastern aspects show significant differences in richness—with a predominance of annual weed species in Spartium —and in above-ground net primary production, probably as a consequence of the nutrients present in the soils. Ruderal annual species under Spartium (Bromus tectorum, Chenopodium album) have higher germination rates in the greenhouse than in the standing vegetation, suggesting they are at potential risk if environmental conditions change.     

Feeder use patterns in group-housed pregnant sows fed with an unprotected electronic sow feeder (Fitmix)

Previous studies on feeder use in group-housed pregnant sows focused on dynamic groups and protected electronic sow feeders (ESF). This study observed 60 pregnant sows, 1st to 8th parity--housed from Day 29 of pregnancy to 1 week before parturition in stable groups of 20 animals, 1 Fitmix feeder per group. Data from 25 nonconsecutive 24-hr feeding cycles showed sows making several visits to the feeder. Literature on conventional ESF indicated shorter daily feeder occupation. Daily feeder occupation per sow decreased over time (p < .001). The study observed maximum feeder activity in the hours following the start of each feeding cycle. During the experiment, there was a relatively stable, quickly established, and maintained feeder order (W > 0.80, p < .001). This highly correlated with dominance rank (r(s) = 0.80, p < .001). High-ranking sows fed earlier and made as many--but longer--visits as low-ranking sows; thus, they occupied the feeder more time every day (p < .01). Although optimization of the feeder efficiency may take several weeks, Fitmix seems an efficient feeding system for medium-size, stable groups of sows.     

Cross-cultural perspectives on physician and lay models of the common cold

We compare physicians and laypeople within and across cultures, focusing on similarities and differences across samples, to determine whether cultural differences or lay-professional differences have a greater effect on explanatory models of the common cold. Data on explanatory models for the common cold were collected from physicians and laypeople in South Texas and Guadalajara, Mexico. Structured interview materials were developed on the basis of open-ended interviews with samples of lay informants at each locale. A structured questionnaire was used to collect information from each sample on causes, symptoms, and treatments for the common cold. Consensus analysis was used to estimate the cultural beliefs for each sample. Instead of systematic differences between samples based on nationality or level of professional training, all four samples largely shared a single-explanatory model of the common cold, with some differences on subthemes, such as the role of hot and cold forces in the etiology of the common cold. An evaluation of our findings indicates that, although there has been conjecture about whether cultural or lay-professional differences are of greater importance in understanding variation in explanatory models of disease and illness, systematic data collected on community and professional beliefs indicate that such differences may be a function of the specific illness. Further generalizations about lay-professional differences need to be based on detailed data for a variety of illnesses, to discern patterns that may be present. Finally, a systematic approach indicates that agreement across individual explanatory models is sufficient to allow for a community-level explanatory model of the common cold.     

Retinoic acid effects on thyroid function of female rats

AimsRetinoic acid is widely used in dermatological treatment and thyroid cancer management; however its possible side-effects on normal thyroid function remains unknown. We aimed to determine the effects of retinoic acid on thyroid function of adult female rats.Main methodsFemale Wistar rats were treated with all-trans-retinoic acid and 13-cis retinoic acid for 14 and 28 days. Then, rats were killed and thyroid function was evaluated.Key findingsSerum T4 and thyrotropin levels remained unchanged, while serum T3 increased in animals treated with all-trans-retinoic acid for 14 days. No changes were observed in hepatic or renal type 1 iodothyronine deiodinase (D1) activities, while thyroid D1 was higher in animals treated for 14 days with all-trans-retinoic acid, which could be related to the increased serum T3 levels. 13-cis retinoic acid increased thyroid iodide uptake after 28 days. These results show effects of retinoic acid treatment on these thyroid proteins: sodium/iodide symporter and deiodinase.SignificanceRetinoic acid is able to interfere with normal thyroid function, increasing thyroid type 1 deiodinase activity, serum T3 levels and sodium/iodide symporter function. However, the effects are time- and retinoic acid isomer-dependent. Since serum thyrotropin levels did not change in any group, the effects observed are probably mediated by a direct retinoic acid effect on the normal thyroid.