CB2 cannabinoid receptor activation is cardioprotective in a mouse model of ischemia/reperfusion

Preventive treatment with cannabinoid agonists has been reported to reduce the infarct size in a mouse model of myocardial ischemia/reperfusion. Here we investigated the possible cardioprotective effect of selective CB₂ cannabinoid receptor activation during ischemia. We performed left coronary artery ligature in C57Bl/6 mice for 30 min, followed by 24 h of reperfusion. Five minutes before reperfusion, mice received intraperitoneal injection of the CB₂ selective agonist JWH-133 (20 mg/kg) or vehicle. Infarct size was assessed histologically and by cardiac troponin I (cTnI) ELISA. Immunohistochemical analysis of leukocyte infiltration, oxidative stress in situ quantification, real-time RT-PCR analysis of inflammatory mediators as well as western blots for kinase phosphorylation was also performed. In addition, we studied chemotaxis and integrin expression of human neutrophils in vitro. JWH-133 significantly reduced the infarct size (I/area at risk: 19.27% ± 1.91) as compared to vehicle-treated mice (31.77% ± 2.7). This was associated with a reduction of oxidative stress and neutrophil infiltration in the infarcted myocardium, whereas activation of ERK 1/2 and STAT-3 was increased. Preinjection of PI3K inhibitor LY294002, MEK 1/2 inhibitor U0126 and JAK-2 inhibitor AG-490 partially abrogated the JWH-133 mediated infarct size reduction. No changes in cardiac CXCL1, CXCL2, CCL3, TNF-α, and ICAM-1 expression levels were found. Furthermore, JWH-133 inhibited the TNF-α induced chemotaxis and integrin CD18/CD11b (Mac-1) upregulation on human neutrophils. Our data suggest that JWH-133 administration during ischemia reduces the infarct size in a mouse model of myocardial ischemia/reperfusion through a direct cardioprotective activity on cardiomyocytes and neutrophils.     

A predicted functional single-nucleotide polymorphism of bone morphogenetic protein-4 gene affects mRNA expression and shows a significant association with cutaneous melanoma in Southern Italian population

Purpose  

An altered expression of bone morphogenetic protein 4 (BMP4) has been found in malignant melanoma cells. We performed an association study to investigate the effect of putative functional single nucleotide polymorphisms (SNPs) of BMP4 on development of cutaneous melanoma (CM).     

Canine cognitive deficit correlates with diffuse plaque maturation and S100β (−) astrocytosis but not with insulin cerebrospinal fluid level

Like humans, canines develop with aging β-amyloid (Aβ) plaques and a progressive cognitive deficit on tasks similar to those used in diagnosis and follow-up of Alzheimer’s disease. Owing to that, dogs are quite unique to investigate the early events taking place in the diffuse Aβ plaque maturation and its relationship with cognitive deficit. The aim of the present investigation was to study the link between the diffuse Aβ plaque maturation and the astro- and microglial reactivity. The involvement of insulin and beta-subunit of S100 protein (S100β) overexpression in the process was also investigated. Aβ plaques were measured and counted in prefrontal cortex of 16 pet dogs of different breeds, weight and sex, classified as control and with a light or severe cognitive deficit. A correlation between canine graded cognitive deficit, diffuse plaque maturation, and S100β (−) astrocytosis, but not with cerebrospinal fluid insulin level, was found that may reflect the very early events of Aβ deposition in Alzheimer’s disease.     

Participation of CYP2C8 and CYP3A4 in the N-demethylation of imatinib in human hepatic microsomes

The receptor activity-modifying proteins (RAMPs) are a family of three single transmembrane proteins that have been identified as accessory proteins to some G-protein-coupled receptors (GPCRs). They can regulate their pharmacology, forward trafficking and recycling, depending on the GPCR. The best characterized receptor complexes formed by RAMPs and GPCRs are the calcitonin peptide family receptors. The association of RAMP1 with the calcitonin receptor-like receptor (CL) constitutes the calcitonin gene-related peptide receptor, whereas RAMP2 or 3 with CL generates adrenomedullin receptors. In this case, the RAMPs substantially alter the pharmacology and trafficking properties of this GPCR. Amylin receptor subtypes are formed from calcitonin receptor (CTR) interactions with RAMPs. Although the RAMPs themselves are not responsive to calcitonin peptide family ligands, there is clear evidence that they participate in ligand binding, although it is still unclear whether this is by directly participating in binding or through allosteric modulation of CL or CTR. A considerable amount of mutagenesis data have now been generated on RAMPs to try and identify the residues that play a role in ligand interactions, and to also identify which residues in RAMPs interact with CL and CTR. This review will focus on RAMP mutagenesis studies with CL, summarizing and discussing the available data in association with current RAMP models and structures. The data reveal key regions in RAMPs that are important for ligand binding and receptor interactions.This article is part of a themed section on Molecular Pharmacology of GPCR. To view the editorial for this themed section visit http://dx.doi.org/10.1111/j.1476-5381.2010.00695.x     

Searching for new animal models of Alzheimer′s disease

The pathophysiology of chronic neurodegenerative diseases, as Alzheimer′s diseases, has remained inaccessible till recently. But this situation is changing quickly. In the past decades, genes causing familiar forms of the disease have been identified and provided the genetic framework for the emerging amyloid hypothesis. On the basis of these findings, engineered mouse models have been developed and have allowed the understanding of crucial information about the pathogenic process. Certain observations obtained by transgenic mice, however, do not easily fit with the simplest version of the amyloid hypothesis. Even if there are transgenic lines that offer robust and relatively faithful reproductions of a subset of Alzheimer′s disease′s features, a mouse model that recapitulates all aspects of the disease has not yet been produced. Several still not completely known factors combine to produce highly variability across transgenic mouse models. Discrepancies in neuropathology and behaviour between transgenic mouse models and human Alzheimer′s disease, and among different transgenic-lines, suggest caution in the interpretation of the results. Here we try to analyze critically some of the information provided by transgenic mice but ascertaining which elements of the neuropathological and behavioural phenotype of these various strains of transgenic mice are relevant to that observed in Alzheimer′s disease continues to be a challenge.     

Side chain cyclization based on serine residues: synthesis, structure, and activity of a novel cyclic analogue of the parathyroid hormone fragment 1-11

The N-terminal region of the parathyroid hormone (PTH) is sufficient to activate the G-protein-coupled PTH receptor 1 (PTHR1). The shortest PTH analogue displaying nanomolar potency is the undecapeptide H-Aib-Val-Aib-Glu-Ile-Gln-Leu-Nle-His-Gln-Har-NH(2) that contains two helix-stabilizing residues (Aib(1,3)). To increase the helical character and proteolytic stability of this linear peptide, we replaced Gln(6,10) with (a) Lys(6) and Glu(10) to introduce a lactam bridge and (b) Ser(6,10) to form a diester bridge upon cross-linking with adipic acid. These cyclopeptides were, respectively, 468-fold less and 12-fold more potent agonists than the linear analogue. Despite their different potencies, all three analogues adopted similar α-helix structures, as shown by NMR and molecular dynamics studies. However, the diester bridge could better mimic the orientation and chemical properties of the side chains of Gln(6) and Gln(10) in the linear PTH analogue than the lactam moiety. This is apparently important for efficient receptor activation and provides further insights into the receptor-bound ligand conformation.     

Thermal Behavior,Structure and Morphology of Propene/Higher 1‐Olefin Copolymers

Summary: Propene‐based random copolymers, synthesized with two isospecific methylaluminoxane‐activated ansa‐zirconocenes and containing even and odd carbon‐numbered higher 1‐olefins, were investigated by DSC and WAXD. Nature and amount of co‐units affect primary and secondary crystallization as well as melting behavior; their role is discussed in the light of microstructural features of the macromolecular chains. The specific role of 1‐olefin comonomers in the development of the γ‐polymorph was assessed by crystallizing selected samples at relatively high temperatures. A good agreement between the fraction of γ‐phase calculated from the deconvolution of the DSC melting endotherms and from WAXD analysis was found. The γ‐phase content nicely correlates with the average length of the isotactic sequences. In the WAXD patterns of some propene/1‐butene copolymers a new reflection at 2θ ? 10°, likely corresponding to the newly discovered trigonal modification of isotactic polypropene, was found.

Room temperature WAXD of copolymer M‐PBu7.5 after isothermal crystallization at two different temperatures.