Up-regulation of the P2Y2 receptor by cytokines in neuronal cells

Abstract only availableAlzheimer's Disease (AD) is characterized by inflammation and neurodegeneration in the brain due to the presence of extracellular amyloid beta (A β) plaques and neurofibrillary tangles. Microglial and astrocyte cells associated with these plaques and tangles have been shown to release cytokines in AD patients, which have a proinflammatory effect on the brain. The P2Y2 receptor (P2Y2R) is a receptor protein that is up-regulated in response to damage or stress in a variety of tissues, including blood vessels and salivary gland epithelium. Recently our laboratory has shown that activation of the P2Y2R enhances α -secretase-dependent amyloid precursor protein (APP) processing. APP is proteolytically processed by β - and γ -secretases to release neurodegenerative A β. Alternatively, APP can be cleaved within the A β domain by α -secretase releasing the non-amyloidogenic product, sAPP α, which has been shown to have neuroprotective properties. Primary neurons have low P2Y2R expression, however, it has been demonstrated that cytokines up-regulate P2Y2R in smooth muscle cells. Therefore, this study will explore if cytokines up-regulate P2Y2R expression in primary rat neurons and in SH-SY5Y human neuroblastoma cells. Primary rat neurons and SH-SY5Y human neuroblastoma cells were plated on glass cover slips 24 or 48 hours with individual treatment, or a combination of, human interleukin-1 β (IL1- β), tumor necrosis factor α (TNF α), and interferon γ (IF γ). P2Y2R activity was measured by increases in intracellular calcium concentration ([Ca2+]i ) in response to the P2Y2R agonist UTP. Results support the hypothesis that P2Y2R is up-regulated by cytokines in neuronal cells. Furthermore, real-time PCR results indicate a two-fold increase in P2Y2R mRNA after cytokine treatment. Therefore, activation of the up-regulated P2Y2R in stressed neurons generates a neuroprotective (sAPP α) rather than neurodegenerative (A β) peptide. These results could have a substantial impact on the understanding and treatment of neurological disorders such as AD.Life Sciences Undergraduate Research Opportunity Progra

TLR3 deficiency exacerbates the loss of epithelial barrier function during genital tract Chlamydia muridarum infection

PROBLEM: Chlamydia trachomatis infections are often associated with acute syndromes including cervicitis, urethritis, and endometritis, which can lead to chronic sequelae such as pelvic inflammatory disease (PID), chronic pelvic pain, ectopic pregnancy, and tubal infertility. As epithelial cells are the primary cell type productively infected during genital tract Chlamydia infections, we investigated whether Chlamydia has any impact on the integrity of the host epithelial barrier as a possible mechanism to facilitate the dissemination of infection, and examined whether TLR3 function modulates its impact. METHOD OF STUDY: We used wild-type and TLR3-deficient murine oviduct epithelial (OE) cells to ascertain whether C. muridarum infection had any effect on the epithelial barrier integrity of these cells as measured by transepithelial resistance (TER) and cell permeability assays. We next assessed whether infection impacted the transcription and protein function of the cellular tight-junction (TJ) genes for claudins1-4, ZO-1, JAM1 and occludin via quantitative real-time PCR (qPCR) and western blot. RESULTS: qPCR, immunoblotting, transwell permeability assays, and TER studies show that Chlamydia compromises cellular TJ function throughout infection in murine OE cells and that TLR3 deficiency significantly exacerbates this effect. CONCLUSION: Our data show that TLR3 plays a role in modulating epithelial barrier function during Chlamydia infection of epithelial cells lining the genital tract. These findings propose a role for TLR3 signaling in maintaining the integrity of epithelial barrier function during genital tract Chlamydia infection, a function that we hypothesize is important in helping limit the chlamydial spread and subsequent genital tract pathology

The P2Y2 nucleotide receptor is an inhibitor of vascular calcification

BACKGROUND AND AIMS: Mutations in the 5'-nucleotidase ecto (NT5E) gene that encodes CD73, a nucleotidase that converts AMP to adenosine, are linked to arterial calcification. However, the role of purinergic receptor signaling in the pathology of intimal calcification is not well understood. In this study, we examined whether extracellular nucleotides acting via P2Y2 receptor (P2Y2R) modulate arterial intimal calcification, a condition highly correlated with cardiovascular morbidity. METHODS: Apolipoprotein E, P2Y2R double knockout mice (ApoE-/-P2Y2R-/-) were used to determine the effect of P2Y2R deficiency on vascular calcification in vivo. Vascular smooth muscle cells (VSMC) isolated from P2Y2R-/- mice grown in high phosphate medium were used to assess the role of P2Y2R in the conversion of VSMC into osteoblasts. Luciferase-reporter assays were used to assess the effect of P2Y2R on the transcriptional activity of Runx2. RESULTS: P2Y2R deficiency in ApoE-/- mice caused extensive intimal calcification despite a significant reduction in atherosclerosis and macrophage plaque content. The ectoenzyme apyrase that degrades nucleoside di- and triphosphates accelerated high phosphate-induced calcium deposition in cultured VSMC. Expression of P2Y2R inhibits calcification in vitro inhibited the osteoblastic trans-differentiation of VSMC. Mechanistically, expression of P2Y2R inhibited Runx2 transcriptional activation of an osteocalcin promoter driven luciferase reporter gene. CONCLUSIONS: This study reveals a role for vascular P2Y2R as an inhibitor of arterial intimal calcification and provides a new mechanistic insight into the regulation of the osteoblastic trans-differentiation of SMC through P2Y2R-mediated Runx2 antagonism. Given that calcification of atherosclerotic lesions is a significant clinical problem, activating P2Y2R may be an effective therapeutic approach for treatment or prevention of vascular calcification

Endothelial Cell-Specific Deletion of P2Y2 Receptor Promotes Plaque Stability in Atherosclerosis-Susceptible ApoE-Null Mice

OBJECTIVE: Nucleotide P2Y2 receptor (P2Y2R) contributes to vascular inflammation by increasing vascular cell adhesion molecule-1 expression in endothelial cells (EC), and global P2Y2R deficiency prevents fatty streak formation in apolipoprotein E null (ApoE-/-) mice. Because P2Y2R is ubiquitously expressed in vascular cells, we investigated the contribution of endothelial P2Y2R in the pathogenesis of atherosclerosis. APPROACH AND RESULTS: EC-specific P2Y2R-deficient mice were generated by breeding VEcadherin5-Cre mice with the P2Y2R floxed mice. Endothelial P2Y2R deficiency reduced endothelial nitric oxide synthase activity and significantly altered ATP- and UTP (uridine 5'-triphosphate)-induced vasorelaxation without affecting vasodilatory responses to acetylcholine. Telemetric blood pressure and echocardiography measurements indicated that EC-specific P2Y2R-deficient mice did not develop hypertension. We investigated the role of endothelial P2Y2R in the development of atherosclerotic lesions by crossing the EC-specific P2Y2R knockout mice onto an ApoE-/- background and evaluated lesion development after feeding a standard chow diet for 25 weeks. Histopathologic examination demonstrated reduced atherosclerotic lesions in the aortic sinus and entire aorta, decreased macrophage infiltration, and increased smooth muscle cell and collagen content, leading to the formation of a subendothelial fibrous cap in EC-specific P2Y2R-deficient ApoE-/- mice. Expression and proteolytic activity of matrix metalloproteinase-2 was significantly reduced in atherosclerotic lesions from EC-specific P2Y2R-deficient ApoE-/- mice. Furthermore, EC-specific P2Y2R deficiency inhibited nitric oxide production, leading to significant increase in smooth muscle cell migration out of aortic explants. CONCLUSIONS: EC-specific P2Y2R deficiency reduces atherosclerotic burden and promotes plaque stability in ApoE-/- mice through impaired macrophage infiltration acting together with reduced matrix metalloproteinase-2 activity and increased smooth muscle cell migration

Central factorials under the Kontorovich-Lebedev transform of polynomials

We show that slight modifications of the Kontorovich-Lebedev transform lead to an automorphism of the vector space of polynomials. This circumstance along with the Mellin transformation property of the modified Bessel functions perform the passage of monomials to central factorial polynomials. A special attention is driven to the polynomial sequences whose KL-transform is the canonical sequence, which will be fully characterized. Finally, new identities between the central factorials and the Euler polynomials are found.Comment: also available at http://cmup.fc.up.pt/cmup/ since the 2nd August 201

Impact of annual praziquantel treatment on urogenital schistosomiasis in a seasonal transmission focus in central Senegal

In Sub-Saharan Africa, urogenital schistosomiasis remains a significant public health problem, causing 150.000 deaths/year with approximately 112 million cases diagnosed. The Niakhar district is a disease hotspot in central Senegal where transmission occurs seasonally with high prevalences. The aim of this study was to determine the effect of annual treatment over 3 years on the seasonal transmission dynamics of S. haematobium in 9 villages in the Niakhar district. Adults and children aged between 5 and 60 years were surveyed from 2011 to 2014. Urine samples were collected door-to-door and examined for S. haematobium eggs at baseline in June 2011, and all participants were treated in August 2011 with PZQ (40 mg/kg). After this initial examination, evaluations were conducted at 3 successive time points from September 2011 to March 2014, to measure the efficacy of the annual treatments and the rates of reinfection. Each year, during the transmission period, from July to November-December, malacological surveys were also carried out in the fresh water bodies of each village to evaluate the infestation of the snail intermediate hosts. At baseline, the overall prevalence of S. haematobium infection was 57.7%, and the proportion of heavy infection was 45.3%, but one month after the first treatment high cure rates (92.9%) were obtained. The overall infection prevalence and proportion of heavy infection intensities were drastically reduced to 4.2% and 2.3%, respectively. The level of the first reinfection in February-March 2012 was 9.5%. At follow-up time points, prevalence levels varied slightly between reinfection and treatment from 9.5% in June 2012 to 0.3% in March 2013, 11.2 in June 2013, and 10.1% April 2014. At the end of the study, overall prevalence was significantly reduced from 57.7% to 10.1%. The overall rate of infested Bulinid snails was reduced after repeated treatment from 0.8% in 2012 to 0.5% in 2013. Repeated annual treatments are suggested to have a considerable impact on the transmission dynamics of S. haematobium in Niakhar, due to the nature of the epidemiological system with seasonal transmission. Thus, to maintain this benefit and continue to reduce the morbidity of urogenital schistosomiasis, other approaches should be integrated into the strategy plans of the National program to achieve the goal of urogenital schistosomiasis elimination in seasonal foci in Senegal

Synthesis, structural characterization, antimicrobial and cytotoxic effects of aziridine, 2-aminoethylaziridine and azirine complexes of copper(II) and palladium(II).

The synthesis, spectroscopic and X-ray structural characterization of copper(II) and palladium(II) complexes with aziridine ligands as 2-dimethylaziridine HNCH2CMe2 (a), the bidentate N-(2-aminoethyl)aziridines C2H4NC2H4NH2 (b) or CH2CMe2NCH2CMe2NH2 (c) as well as the unsaturated azirine NCH2CPh (d) are reported. Cleavage of the cyclometallated Pd(II) dimer [μ-Cl(C6H4CHMeNMe2-C,N)Pd]2 with ligand a yielded compound [Cl(NHCH2CMe2)(C6H4CHMe2NMe2-C,N)Pd] (1a). The reaction of the aziridine complex trans-[Cl2Pd(HNC2H4)2] with an excess of aziridine in the presence of AgOTf gave the ionic chelate complex trans-[(C2H4NC2H4NH2-N,N′)2Pd](OTf)2 (2b) which contains the new ligand b formed by an unexpected insertion and ring opening reaction of two aziridines (“aziridine dimerization”). CuCl2 reacted in pure HNC2H4 or HNCH2CMe2 (b) again by “dimerization” to give the tris-chelated ionic complex [Cu(C2H4NC2H4NH2-N,N′)3]Cl2 (3b) or the bis-chelated complex [CuCl(C2H2Me2NC2H2Me2NH2-N,N′)2]Cl (4c). By addition of 2H-3-phenylazirine (d) to PdCl2, trans-[Cl2Pd(NCH2CPh)2] (5d) was formed. All new compounds were characterized by NMR, IR and mass spectra and also by X-ray structure analyses (except 3b). Additionally the cytotoxic effects of these complexes were examined on HL-60 and NALM-6 human leukemia cells and melanoma WM-115 cells. The antimicrobial activity was also determined. The growth of Gram-positive bacterial strains (S. aureus, S. epidermidis, E. faecalis) was inhibited by almost all tested complexes at the concentrations of 37.5–300.0 μg mL−1. However, MIC values of complexes obtained for Gram-negative E. coli and P. aeruginosa, as well as for C. albicans yeast, mostly exceeded 300 μg mL−1. The highest antibacterial activity was achieved by complexes 1a and 2b. Complex 2b also inhibited the growth of Gram-negative bacteria. Graphical abstract: Synthesis, structural characterization, antimicrobial and cytotoxic effects of aziridine, 2-aminoethylaziridine and azirine complexes of copper(ii) and palladium(ii

D 5.3.2.2. Initial report on sensory and African consumer acceptance for Group 2. Project AFTER “African Food Tradition rEvisited by Research”.

The sensory profile and consumer acceptance of Lanhouin, a traditional salted fermented fish made from two types of fish (king fish and cassava fish) and three different fermentation processes (aerobic, semi aerobic and anaerobic condition) were explored. Traditional Lanhouin were sensorially different, with king fish Lanhouin having the highest smell (score of putrid odour) and cassava fish Lanhouin being the whitish. Three classes of consumers' behaviour essentially, those who like all the Lanhouin samples (Lanhouin likers; 35 % of consumers), those who does not prefer king fish Lanhouin from anaerobic fermentation (Lanhouin KFAN dislikers; 37 %) and those who does not like cassava fish Lanhouin from aerobic fermentation (Lanhouin CFA dislikers; 28 %) were identified. Consumers' acceptance was significantly associated with size of fish, whitish colour, dried fish odour and ammoniacal odour. Considering consumers' preference, Lanhouin from king fish with semi aerobic fermentation and Lanhouin from cassava fish (except for aerobic fermentation) were more accepted and could be interesting for reengineering. The similarity in tasting attributes for cassava fish and king fish when used for cooked rice will be helpful to design the form in which Lanhouin can be marketed for large acceptance Considering Kong, no prior work has explored sensory tasting. Four samples were produced for the sensory test: Kong was smoked with wood only and or with wood/coconut. For the sensory evaluation, smoked kong only and smoked kong in sauce and were scored by a semitrained sensory panel. Then, 109 consumers were interviewed. Acceptability was only evaluated on smoked kong. The results show that all four samples were quite distinctive from each other but not when in sauce. Most of the Kong products were on average acceptable but moist kong had a better acceptability than dried kong samples submitted to the same processes of smoking. Consumers were segmented into two three groups as: “indifferent likers” (27%), “dry kong dislikers” (15%), and “dry Kong less preferred” (59%). Clusters really differed in terms of professional activity in fish sector but did not significantly differ in the frequency of consumption. Therefore it appears that, smoked Kong is a main product of consumption in the Senegalese market. Since more than 50% of consumers consumed smoked Kong at least once a week, it appears that Kong is a main product of consumption with a strong potential for the Senegalese market but also for the international market. (Résumé d'auteur

D 5.3.2.3. Initial report on sensory and African consumer acceptance for Group 3. Project AFTER “African Food Tradition rEvisited by Research”

Sensory evaluation and consumer testing of Hibiscus, Baobab and Jaabi (Yaabande) was undertaken with African and EU consumers in Senegal and Cameroon. The sensory profile and consumer acceptance of Bissap (Hibiscus sabdariffa L.) drink (commercial and traditional) made from infusions of red calices of either local or Sudanese origin, pure or mixed, and either as syrup or a juice (infusion) was explored. There were significant correlations between the sensory perceptions of the drink by panellists. The acceptability of the drink was tested by two consumer groups: Europeans (n=60) and Africans (n=100) in Dakar. Three classes of behaviour from the consumers were identified. There were a) those who preferred syrup (syrup likers; 43% of consumers) b) those who prefer juices (juice likers; 36% of consumers) and c) those who preferred all of the samples (indifferent likers; 21% of consumers). The liking by African and EU consumers was similar but European consumers were more likely to prefer syrup, consume bissap the least often and purchase bissap juice in bottles. African consumers were more likely to prefer juice, consume bissap more frequently and purchase it in sachets. Both groups preferred to purchase natural bissap drinks rather than ones with added flavour. The sensory characteristics important to each class of consumers differed. There were significant correlations between acceptability, bissap taste for the cluster of the group of the juice likers. There were significant correlations between acceptability, sweet taste, acidic taste for the syrup likers. The study shows that the distinctions between the acceptability groups are very clear from a sensory perspective. The market study should take these preference groups into account when launching Hibiscus drinks on a new market. The sensory profile of seven samples of bouye (baobab) drinks (syrup and juice) was evaluated by 17 panellists. Consumer testing was investigated at four different locations in Dakar using the central location metho. These were the following: ESP high school (n=32); Grand Yoff (n=36), Guediawaye (n=12), and Gueule Tapée (n=26) areas. The acceptability of the drink was tested by African consumers (mainly Senegalese) (n=104) in Dakar who tested five different drinks from the seven first samples. A cluster analysis demonstrated that consumers behave differently with respect to acceptability. Three classes of behaviour from the consumers were identified. There were a) those who clearly preferred the juice from FWS (27% of consumers) b) those who preferred all of the samples (indifferent likers) (31% of consumers), c) those who liked the juices but not the syrups (juice likers) (42% of consumers). With respect to acceptance, the reengineering approach appears to have two options being products suited to a) milk taste or b) taste and odour characteristic of baobab with a good concentration for the two products. The reengineering approach should also explore ways to increase the shelf life with optimum quality regarding the sensory attributes and consumer acceptance. Optimizing scales for pasteurization shall be conducted in order to have a product without caramel smell. Sensory profile and consumer acceptability was undertaken for Jaabi, a wild fruit commonly used in savannah region of Africa, and its processed product, a cake locally called: Yaabande. The study took in the northern part of Cameroon through collection of two varieties of Jaabi (Dakamji and Lamouji) from four origins (Garoua, Maroua, Mokolo, Mora), and processed Yaabande from these samples, using three baking techniques (Sun baking, vapour cooking, under earth stewing). Sensory and consumers tests indicated that the taste of the products constitutes the main criteria for consuming the products. The Dakamji variety of Jaabi appeared more homogenous whatever the origin than the Lamouji variety. Meanwhile, all samples were acceptable at comparable level corresponding to pleasant character. With respect to acceptance, the main option of reengineering approach is based on standardisation of process procedures in order to guarantee the taste of the products. Inclusion of therapeutic aspects of the products may be coupled to this option for market development. (Résumé d'auteur

Allogenic hematopoietic stem cell transplantation after reduced intensity conditioning regimen for elderly patients (60 years and older) with hematologic malignancies using unrelated donors: a retrospective study for the French society for stem cell transplantation (SFGM-TC)

Peer reviewe