Pivaloylcodeine,a new codeine derivative,for the inhibition of morphine glucuronidation. An in vitro study in the rat

We have previously found that phenanthrenic opioids, including codeine, modulate morphine glucuronidation in the rat. Here codeine and five of its derivatives were compared in their effects on the synthesis of morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) from morphine by rat liver microsomal preparations, and by primary cultures of rat hepatocytes previously incubated for 72 h with either codeine or its derivatives. Acetylcodeine and pivaloylcodeine shared the capability of the parent compound of inhibiting the synthesis of M3G by liver microsomes through a noncompetitive mechanism of action. Their IC₅₀ were 3.25, 2.27, and 4.32 μM, respectively. Dihydrocodeine, acetyldihydrocodeine, and lauroylcodeine were ineffective. In all the experimental circumstances M6G was undetectable in the incubation medium. In primary hepatocyte cultures codeine only inhibited M3G formation, but with a lower efficacy than that observed with microsomes (IC₅₀ 20.91 vs 4.32 μM). Preliminary results show that at micromolar concentrations codeine derivatives exhibit a low rate of affinity for μ opiate receptors. In conclusion, acetyl and pivaloyl derivatives of codeine noncompetitively inhibit liver glucuronidation of morphine interacting with microsomes. This study further strengths the notion that phenanthrenic opioids can modulate morphine glucuronidation independently from their effects on μ opiate receptors.     

Vessel length and conductivity of Ulmus branches: ontogenetic changes and relation to resistance to Dutch elm disease

Changes in age of the hydraulic architecture of Ulmus minor and U. minor × U. pumila juvenile wood were studied and related to tolerance to Dutch elm disease (DED). The xylem vessel dimensions and the conductivity to air of 2- to 7-year-old branches were analyzed and quantified. No obvious differences in vessel length distribution and conductivity were found to explain differences in DED tolerance among the U. minor clones, or, at the taxon level, the higher DED tolerance of U. minor × U. pumila. Among the U. minor clones, the more susceptible one had wider vessels and a higher maximum vessel diameter than the more tolerant clone. Relations between vessel lengths, vessel diameters and branch sizes were highly significant, and varied between taxa. The diameter and length of vessels increased with age, and average values stabilized 1–2 years earlier for U. minor than for U. minor × pumila. Mean maximum vessel length was significantly higher in U. minor and increased more with age and maximum vessel diameter than in U. minor × pumila. With each 0.2 m increase in height up the stem, conductivities for U. minor and U. minor × U. pumila decreased by 59 and 50 %, respectively, probably due to shortening of the vessels. The implications of xylem structure for the means of pathogen movement and resistance to DED are discussed.     

Which are, what is their status and what can we expect from ecosystem services provided by Spanish rivers and riparian areas?

By applying the framework proposed by Millennium Ecosystem Assessment, we analysed the current state and trends of 20 ecosystem services provided by Spanish rivers and riparian areas using 139 indicators. We compared the obtained results with the Europe and UK assessment. It is the first document that attempts to analyse the importance of services provided by Spanish rivers and riparian areas, and it forms part of the evaluation carried out for the Spanish Millennium Ecosystem Assessment. Among the provisioning services, freshwater, hydropower energy and genetic resources were classified as high importance, and water regulation and self-purification capacity, and natural hazard mitigation are among the regulating services, with landscape-aesthetic values and recreation and ecotourism featuring among the cultural services. About 61 % of the assessed ecosystem services are currently declining or degrading, but are higher than the percentage calculated for Europe (45 %) or for the UK (53 %). All regulating services are degrading, especially water regulation, natural hazard mitigation, soil formation and fertility and biological control, and the cultural services related to rural populations. Likewise, the biodiversity of Spanish aquatic ecosystems is decreasing rapidly. Land use changes and overexploitation of biological and mineral raw materials have been the main direct drivers of change in Spanish rivers and riparian zones, and relate directly to increase urbanised areas and irrigated agriculture. Finally, we draw some considerations on alternative models for aquatic ecosystems management which maintain aquatic ecosystem services and their biodiversity.     

Cellulase Production in Semisolid Cultures of Trichoderma viride

Cellulase was produced by Trichoderma viride in semisolid cultures of rice bran, rice straw and rice hulls. T. viride QM 9414 generally produced higher cellulolytic activity on CM-cellulose (C_x activity) using rice bran-rice hull mixture (2:1 w/w) as substrate compared to strains ITCC 1433 and D 4014. It showed higher C_x activity on rice bran-rice straw mixtures than on rice bran-rice hull mixtures. Maximal extraction of the enzyme from mold bran was obtained with 0.05 m sodium citrate buffer, pH 3.5.     

Interactions of Methicillin Resistant Staphylococcus aureus USA300 and Pseudomonas aeruginosa in Polymicrobial Wound Infection

Understanding the pathology resulting from Staphylococcus aureus and Pseudomonas aeruginosa polymicrobial wound infections is of great importance due to their ubiquitous nature, increasing prevalence, growing resistance to antimicrobial agents, and ability to delay healing. Methicillin-resistant S. aureus USA300 is the leading cause of community-associated bacterial infections resulting in increased morbidity and mortality. We utilized a well-established porcine partial thickness wound healing model to study the synergistic effects of USA300 and P. aeruginosa on wound healing. Wound re-epithelialization was significantly delayed by mixed-species biofilms through suppression of keratinocyte growth factor 1. Pseudomonas showed an inhibitory effect on USA300 growth in vitro while both species co-existed in cutaneous wounds in vivo. Polymicrobial wound infection in the presence of P. aeruginosa resulted in induced expression of USA300 virulence factors Panton-Valentine leukocidin and α-hemolysin. These results provide evidence for the interaction of bacterial species within mixed-species biofilms in vivo and for the first time, the contribution of virulence factors to the severity of polymicrobial wound infections.     

Poor Immune Reconstitution in HIV-Infected Patients Associates with High Percentage of Regulatory CD4+ T Cells

CD4⁺ regulatory T cells (Tregs) are essential for the maintenance of the immune system''s equilibrium, by dampening the activation of potential auto-reactive T cells and avoiding excessive immune activation. To correctly perform their function, Tregs must be maintained at the right proportion with respect to effector T cells. Since this equilibrium is frequently disrupted in individuals infected with the human immunodeficiency virus (HIV), we hypothesize that its deregulation could hamper immune reconstitution in patients with poor CD4⁺ T cell recovery under highly active antiretroviral therapy (HAART). We analysed Tregs percentages amongst CD4⁺ T cells in 53 HIV-infected patients under HAART, with suppression of viral replication and distinct levels of immune reconstitution. As controls, 51 healthy individuals were also analysed. We observed that amongst the patients with Nadir values (the lowest CD4⁺ T cell counts achieved) <200 cells/µL, the individuals with high Tregs percentages (≥10% of total CD4⁺ T cells) had the worse CD4⁺ T cell reconstitution. In accordance, the well-described direct correlation between the Nadir value and CD4⁺ T cell reconstitution is clearly more evident in individuals with high Tregs proportions. Furthermore, we observed a strong negative correlation between Tregs percentages and CD4⁺ T cell recovery among immunological non-responder HIV⁺ individuals. All together, this work shows that high Tregs frequency is an important factor associated with sub-optimal CD4⁺ T cell recovery. This is particularly relevant for immunological non-responders with low Nadir values. Our results suggest that the Tregs proportion might be of clinical relevance to define cut-offs for HAART initiation.     

An Overview of Ten Italian Horse Breeds through Mitochondrial DNA

BackgroundThe climatic and cultural diversity of the Italian Peninsula triggered, over time, the development of a great variety of horse breeds, whose origin and history are still unclear. To clarify this issue, analyses on phenotypic traits and genealogical data were recently coupled with molecular screening.MethodologyTo provide a comprehensive overview of the horse genetic variability in Italy, we produced and phylogenetically analyzed 407 mitochondrial DNA (mtDNA) control-region sequences from ten of the most important Italian riding horse and pony breeds: Bardigiano, Esperia, Giara, Lipizzan, Maremmano, Monterufolino, Murgese, Sarcidano, Sardinian Anglo-Arab, and Tolfetano. A collection of 36 Arabian horses was also evaluated to assess the genetic consequences of their common use for the improvement of some local breeds.ConclusionsIn Italian horses, all previously described domestic mtDNA haplogroups were detected as well as a high haplotype diversity. These findings indicate that the ancestral local mares harbored an extensive genetic diversity. Moreover, the limited haplotype sharing (11%) with the Arabian horse reveals that its impact on the autochthonous mitochondrial gene pools during the final establishment of pure breeds was marginal, if any. The only significant signs of genetic structure and differentiation were detected in the geographically most isolated contexts (i.e. Monterufolino and Sardinian breeds). Such a geographic effect was also confirmed in a wider breed setting, where the Italian pool stands in an intermediate position together with most of the other Mediterranean stocks. However, some notable exceptions and peculiar genetic proximities lend genetic support to historical theories about the origin of specific Italian breeds.