n−3 polyunsaturated N-acylethanolamines are CB2 cannabinoid receptor-preferring endocannabinoids

Anandamide, the first identified endogenous cannabinoid and TRPV1 agonist, is one of a series of endogenous N-acylethanolamines, NAEs. We have generated novel assays to quantify the levels of multiple NAEs in biological tissues and their rates of hydrolysis through fatty acid amide hydrolase. This range of NAEs was also tested in rapid response assays of CB₁, CB₂ cannabinoid and TRPV1 receptors. The data indicate that PEA, SEA and OEA are not endocannabinoids or endovanilloids, and that the higher endogenous levels of these metabolites compared to polyunsaturated analogues are a correlate of their slow rates of hydrolysis. The n?6 NAEs (AEA, docosatetraenoyl and docosapentaenoyl derivatives) activated both CB₁ and CB₂ receptors, as well as TRPV1 channels, suggesting them to be ‘genuine’ endocannabinoids and ‘endovanilloids’. The n?3 NAEs (eicosapentaenoyl, docosapentaenoyl and docosahexaenoyl derivatives) activated CB₂ receptors and some n?3 NAEs (docosapentaenoyl and docosahexaenoyl derivatives) also activated TRPV1 channels, but failed to activate the CB₁ receptor. We hypothesise that the preferential activation of CB₂ receptors by n?3 PUFA NAEs contributes, at least in some part, to their broad anti-inflammatory profile.     

Pyoverdine and histicorrugatin-mediated iron acquisition in <Emphasis Type="Italic">Pseudomonas thivervalensis</Emphasis>

The genome of Pseudomonas thivervalensis LMG 21626^T has been sequenced and a genomic, genetic and structural analysis of the siderophore mediated iron acquisition was undertaken. Pseudomonas thivervalensis produces two structurally new siderophores, pyoverdine PYO_thi which is typical for P. thivervalensis strains and a closely related strain, and the lipopeptidic siderophore histicorrugatin which is also detected in P. lini. Histicorrugatin consists out of an eight amino acid long peptide which is linked to octanoic acid. It is structurally related to the siderophores corrugatin and ornicorrugatin. Analysis of the proteome for TonB-dependent receptors identified 25 candidates. Comparison of the TonB-dependent receptors of P. thivervalensis with the 17 receptors of its phylogenetic neighbor, P. brassicacearum subsp. brassicacearum NFM 421, showed that NFM 421 shares the same set of receptors with LMG 21626^T, including the histicorrugatin receptor. An exception was found for their cognate pyoverdine receptor which can be explained by the observation that both strains produce structurally different pyoverdines. Mass analysis showed that NFM 421 did not produce histicorrugatin, but the analogue ornicorrugatin. Growth stimulation assays with a variety of structurally distinct pyoverdines produced by other Pseudomonas species demonstrated that LMG 21626^T and NFM 421 are able to utilize almost the same set of pyoverdines. Strain NFM 421 is able utilize two additional pyoverdines, pyoverdine of P. fluorescens Pf0–1 and P. citronellolis LMG 18378^T, these pyoverdines are probably taken up by the FpvA receptor of NFM 421.     

Physiological studies on Biomphalaria alexandrina and Bulinus truncatus,the snail vectors of Schistosommiasis

Summary The Warburg's manometric technique was used to measure the rate of oxygen consumption of the second generation of laboratory-reared snails, Biomphalaria alexandrina and Bulinus truncatus at two temperatures of 25° and 30°C. The individual weight of the experimental snails ranged between 40 and 78 mg for B. alexandrina, between 60 and 90 mg for B. truncatus.At 25°C, the uninfected snails B. alexandrina consumed oxygen at an average rate of 0.096 ± 0.020 ml/g wet wt/hr. The rate of oxygen consumption increased to an average of 0.147 ± 0.008 ml/g wet wt/hr for uninfected snails maintained at 30°C (about 53 per cent increase). The average RW value for uninfected snails maintained at 25°C was 0.80.The snail Bulinus truncatus showed higher oxygen requirements than the snail Biomphalaria alexandrina. At 25°C, it consumed oxygen at an average rate of 0.124 ± 0.016 ml/g wet wt/hr. At 30°C, the rate of oxygen consumption reached a value of 0.220 + 0.006 ml/g wet wt/hr. The average RQ for Bulinus truncatus maintained at 25°C was 0.87.The rate of oxygen consumption of the schistosome — infected Biomphalaria alexandrina snails, maintained at 25°C decreased to an average rate of 0.059 ± 0.010 ml/g wet wt/hr, (an average of 39 per cent decrease). The respiratory quotient (RQ) also decreased to an average value of 0.58. Further research is suggested to clarify the metabolism of both schistosome-infected and uninfected snails.Read at the Ist African Symposium on Bilharziasis, Cairo Egypt, U.A.R., February, 1969.From the Laboratory of Bilharziasis Research, National Research Centre, Dokki, Egypt, U.A.R.     

Biodegradable fumarate-based polyHIPEs as tissue engineering scaffolds

PolyHIPEs show great promise as tissue engineering scaffolds due to the tremendous control of pore size and interconnectivity afforded by this technique. Highly porous, fully biodegradable scaffolds were prepared by polymerization of the continuous phase of high internal phase emulsions (HIPEs) containing the macromer poly(propylene fumarate) (PPF) and the cross-linker propylene fumarate diacrylate (PFDA). Toluene was used as a diluent to reduce the viscosity of the organic phase to enable HIPE formation. A range of polyHIPE scaffolds of different pore sizes and morphologies were generated by varying the diluent concentration (40-60 wt %), cross-linker concentration (25-75 wt %), and macromer molecular weight ( M n = 800-1000 g/mol). Although some formulations resulted in macroporous monoliths (pore diameter >500 microm), the majority of the polyHIPEs studied were rigid, microporous monoliths with average pore diameters in the range 10-300 microm. Gravimetric analysis confirmed the porosity of the microporous monoliths as 80-89% with most scaffolds above 84%. These studies demonstrate that emulsion templating can be used to generate rigid, biodegradable scaffolds with highly interconnected pores suitable for tissue engineering scaffolds.     

Early trafficking events of Mycobacterium ulcerans within Naucoris cimicoides

The severe skin-destructive disease caused by Mycobacterium ulcerans, named Buruli ulcer, is the third most important mycobacterial disease in humans after tuberculosis and leprosy. Recently we demonstrated that M. ulcerans could colonize the salivary glands of the water bug, Naucoris cimicoides. In this study, we report that M. ulcerans may be delivered from the digested prey aspirate to the coelomic cavity via a unique headspace, the head capsule (HC). During the infected meal, we observed that M. ulcerans clusters adhered to the stylets that were retracted in the HC at the end of the meal. M. ulcerans was able to translocate from the HC to the coelomic cavity where it is phagocytosed by the plasmatocytes. These cells are subverted as shuttle cells and deliver M. ulcerans to the salivary glands. At this early stage of its parasitic life style, two other important features of M. ulcerans can be documented: first, mycolactone is not required for translocation of M. ulcerans into the HC, in contrast to the next step, colonization of the salivary glands; second, M. ulcerans clusters bind a member of the serpin protein family present in the salivary gland homogenate.     

Stable isotope probing of bacterial community structure and gene expression in the rhizosphere of Arabidopsis thaliana

The rhizosphere is an active compartment where plant and microorganisms establish a molecular dialogue. In this study, we analysed the impact of Arabidopsis thaliana on bacterial community structure and the expression of certain beneficial genes using DNA- and mRNA-SIP in the rhizosphere of plantlets grown under (13)CO(2) for 13, 21 and 27 days. DNA- and rRNA-SIP revealed changes in bacterial communities inhabiting the rhizosphere soil that were probably related to modification of root exudates, while root-colonizing populations were maintained over time suggesting their metabolic versatility and adaptation. The impact of the plant via root exudates on the expression of the noncoding RNAs rsmZ, acdS gene encoding 1-aminocyclopropane-1-carboxylate deaminase and nosZ gene encoding nitrous oxide reductase, in the root-adhering soil and on the roots of A. thaliana was determined using mRNA-SIP. Results showed that these genes were present and expressed by bacteria inhabiting roots and by those that derive nutrients from the breakdown of organic matter in soils or from root exudates. The expression of rsmZ under natural conditions indicates the importance of noncoding RNAs in bacterial adaptation to their ecological niches.     

Community onsite treatment of cold strong sewage in a UASB-septic tank

Two community onsite UASB-septic tanks namely R1 and R2 were operated under two different HRT (2 days for R1 and 4 days for R2) in parallel over a year and monitored over the cold half of the year. During the monitoring period, the sewage was characterised by a high COD(tot) of 905mg/l with a high fraction of COD(ss), viz. about 43.7%, and rather low temperature of 17.3 degrees C. The achieved removal efficiencies in R1 and R2 for COD(tot), COD(sus), COD(col), COD(dis), BOD(5) and TSS were "51%, 83%, 20%, 24%, 45% and 74%" and "54%, 87%, 10%, 28%, 49% and 78%", respectively. The difference in the removal efficiencies of those parameters in R1 and R2 is marginal and was only significant (p<0.05) for COD(sus). The sludge filling period of the reactors is expected to be 4-7 years. In view of that, the UASB-septic tank system is a robust and compact system as it can be adequately designed in Palestine at 2 days HRT.     

Effect of water stress by Polyethylene Glycol 8000 and Sodium Chloride on germination of Ephedra alata Decne seeds

Ephedra alata Decne is a perennial shrub and it is a very effective sand-binder. In Saudi Arabia, the species is associated with sand dunes formation, especially the mobile, non-saline and low moisture content ones. Its geographical distribution in Saudi Arabia includes the Northern, Eastern and Central regions. The aims of this study were to determine the effects of temperature, water potential and Sodium Chloride on germination of E. alata. Seeds were collected from King Khalid Centre of Wildlife Research and Development at Thumama (80 km north east of Riyadh), Saudi Arabia. Seeds were germinated at four alternating temperature regimes (8/22; 9/23; 13/27 and 18/35 °C). Seeds were also germinated under stress of aqueous Polyethylene Glycol (PEG) solutions mixed to create water potentials of 0; −0.3; −0.6; −1.2 and −1.5 MPa. Seed were also germinated in Sodium Chloride solutions of 0, 0.05, 0.1, 0.2 and 0.3 mol l⁻¹. Optimum germination was attained at 13/28 °C that corresponds to temperatures prevailing during spring time. Seeds germinated in Polyethylene Glycol solutions exhibited significantly lower germination than control especially when water potential fell below −0.3 MPa. Germination was also negatively affected by 0.1 mol l⁻¹ Sodium Chloride solution or above. Results indicated that the germination temperature responses of the nondormant seeds synchronize the event of germination with the season when environmental conditions are more favorable for subsequent growth and seedling establishment. Germination was also sensitive to both water potential and salinity.     

Neuropharmacological screening of two 1,5-benzodiazepine compounds in mice

This work investigates whether the two 1,5-benzodiazepine compounds: 4-(2-hydroxyphenyl)-1,5-benzodiazepin-2-one (RG0501) and Benzopyrano [4,3-c] 1,5-benzodiazepine (RG0502) have any neuropharmacological activities. Diazepam and Flunitrazepam were used as drug references. The investigational 1,5-BDZ were tested in vivo for potentiating hexobarbital-induced sleep and pentylenetetrazole (PTZ)-induced seizures. Our study demonstrated that the increase of sleep duration was significantly higher with RG0501 as compared to RG0502. However, RG0502 anticonvulsant effect was more pronounced than that of RG0501 in the range dose of 6.25-37.5 mg.kg^-1. From the 50 mg.kg^-1 dose, RG0502 offered a protection against clonic-tonic seizures as well as lethality (p ≤ 0.05). The results showed that the required doses to obtain a pharmacological activity were more than those of the references. This difference could be related to the lack of specific substituants responsible for the pharmacological activity in the tested compounds.