Toxoplasmal and Toxocaral Infections: A Clinical Investigation into Their Relationship

An investigation was carried out to determine whether a relationship existed between infections with Toxoplasma gondii and Toxocara canis. Antibodies were sought by the toxoplasma dye test and the toxocara skin sensitivity test. Sixty-seven patients were examined in the United Kingdom; 38 had positive toxoplasma dye tests, two of these being toxocara-positive; and 29 had negative dye tests, six of them being toxocara-positive. From the total of eight toxocarapositive patients antibodies to toxoplasma were detected in two, an incidence no greater than that expected for a normal population.Sera from 60 toxocara-positive African patients were examined; 20 possessed antibodies to Toxoplasma. Analysis according to age group or geographical location indicated that this incidence was no greater than that expected for patients without evidence of toxocaral infection.This study therefore showed no causal or clinically important relationship between toxoplasmal and toxocaral infections.     

Repeated oral administration of capsaicin increases anxiety-like behaviours with prolonged stress-response in rats

This study was conducted to examine the psycho-emotional effects of repeated oral exposure to capsaicin, the principal active component of chili peppers. Each rat received 1 mL of 0.02% capsaicin into its oral cavity daily, and was subjected to behavioural tests following 10 daily administrations of capsaicin. Stereotypy counts and rostral grooming were significantly increased, and caudal grooming decreased, in capsaicin-treated rats during the ambulatory activity test. In elevated plus maze test, not only the time spent in open arms but also the percent arm entry into open arms was reduced in capsaicin-treated rats compared with control rats. In forced swim test, although swimming duration was decreased, struggling increased in the capsaicin group, immobility duration did not differ between the groups. Repeated oral capsaicin did not affect the basal levels of plasma corticosterone; however, the stress-induced elevation of plasma corticosterone was prolonged in capsaicin treated rats. Oral capsaicin exposure significantly increased c-Fos expression not only in the nucleus tractus of solitarius but also in the paraventricular nucleus. Results suggest that repeated oral exposure to capsaicin increases anxiety-like behaviours in rats, and dysfunction of the hypothalamic-pituitary-adrenal axis may play a role in its pathophysiology.     

Massively Parallel Sequencing Reveals the Complex Structure of an Irradiated Human Chromosome on a Mouse Background in the Tc1 Model of Down Syndrome

Down syndrome (DS) is caused by trisomy of chromosome 21 (Hsa21) and presents a complex phenotype that arises from abnormal dosage of genes on this chromosome. However, the individual dosage-sensitive genes underlying each phenotype remain largely unknown. To help dissect genotype – phenotype correlations in this complex syndrome, the first fully transchromosomic mouse model, the Tc1 mouse, which carries a copy of human chromosome 21 was produced in 2005. The Tc1 strain is trisomic for the majority of genes that cause phenotypes associated with DS, and this freely available mouse strain has become used widely to study DS, the effects of gene dosage abnormalities, and the effect on the basic biology of cells when a mouse carries a freely segregating human chromosome. Tc1 mice were created by a process that included irradiation microcell-mediated chromosome transfer of Hsa21 into recipient mouse embryonic stem cells. Here, the combination of next generation sequencing, array-CGH and fluorescence in situ hybridization technologies has enabled us to identify unsuspected rearrangements of Hsa21 in this mouse model; revealing one deletion, six duplications and more than 25 de novo structural rearrangements. Our study is not only essential for informing functional studies of the Tc1 mouse but also (1) presents for the first time a detailed sequence analysis of the effects of gamma radiation on an entire human chromosome, which gives some mechanistic insight into the effects of radiation damage on DNA, and (2) overcomes specific technical difficulties of assaying a human chromosome on a mouse background where highly conserved sequences may confound the analysis. Sequence data generated in this study is deposited in the ENA database, Study Accession number: ERP000439.     

Stress-inducible transgenic nematodes as biomonitors of soil and water pollution

This paper reviews the current status of nematodes with stress-inducible transgenes as biosensors responsive to a range of external stressors, e.g., soil or water pollution, microwave radiation or immunological attack. TransgenicCaenorhabditis elegans carrying reporter genes under heat shock promoter control express reporter products only under stressful conditions. Although relatively insensitive to single metal ions, these worms respond to complex mixtures present in metal-contaminated watercourses and to laboratory mixtures containing similar constituents, but not to any of their components singly at comparable concentrations. Responses to metal mixtures are enhanced by a non-ionic surfactant, Pluronic F-127. Metals taken up by food bacteria and insoluble metal carbonates can also evoke stress responses, both in soil and aqueous media. However, high concentrations of added metals are needed to induce clear-cut responses in soil, owing to metal sorption onto clays and organic matter. Transgenic worms are also stressed by exposure to microwave radiation; pulsed signals generate responses that diminish markedly with distance from the source. Finally, stress responses are inducible by anti-epicuticle antisera and complement, suggesting that immune attack can also activite the heat shock system. The development of rapid microplate toxicity assays based on transgenic nematodes is discussed.     

Measurement of monomer-oligomer distributions via fluorescence moment image analysis

We present higher-order moment analysis of fluorescence intensity fluctuations from individual laser scanning microscopy images applied to study monomer-oligomer distributions. We demonstrate that the number densities and brightness ratios of a mixed population of monomers and oligomers can be determined by analyzing higher-order moments of the fluorescence intensity fluctuations from individual images for specific ranges of densities and particle brightness ratios. Computer simulations and experiments with fluorescent microspheres and cells were performed to illustrate the detection limits and accuracy of this statistical approach. The simulation results show that the concentration of the dimer or oligomer population should be less than or equal to the monomeric concentration for the method to provide accurate results, and that the upper density detection limit of the population of monomers is one order-of-magnitude higher than the concentration of the oligomers. We implemented this technique to resolve two populations of fluorescent microspheres with different brightness ratios and we also applied the moment-analysis method to examine the distribution of aggregation states of PDGF-beta receptors in human fibroblast cells. The method was able to resolve a tetrameric population of the PDGF-beta receptors relative to the background distribution of nonspecifically bound fluorophore.     

Selective inhibition of ATPase activity during contraction alters the activation of p38 MAP kinase isoforms in skeletal muscle

Muscle contractions strongly activate p38 MAP kinases, but the precise contraction‐associated sarcoplasmic event(s) (e.g., force production, energetic demands, and/or calcium cycling) that activate these kinases are still unclear. We tested the hypothesis that during contraction the phosphorylation of p38 isoforms is sensitive to the increase in ATP demand relative to ATP supply. Energetic demands were inhibited using N‐benzyl‐p‐toluene sulphonamide (BTS, type II actomyosin) and cyclopiazonic acid (CPA, SERCA). Extensor digitorum longus muscles from Swiss Webster mice were incubated in Ringer's solution (37°C) with or without inhibitors and then stimulated at 10 Hz for 15 min. Muscles were immediately freeze‐clamped for metabolite and Western blot analysis. BTS and BTS + CPA treatment decreased force production by 85%, as measured by the tension time integral, while CPA alone potentiated force by 310%. In control muscles, contractions resulted in a 73% loss of ATP content and a concomitant sevenfold increase in IMP content, a measure of sustained energetic imbalance. BTS or CPA treatment lessened the loss of ATP, but BTS + CPA treatment completely eliminated the energetic imbalance since ATP and IMP levels were nearly equal to those of non‐stimulated muscles. The independent inhibition of cytosolic ATPase activities had no effect on contraction‐induced p38 MAPK phosphorylation, but combined treatment prevented the increase in phosphorylation of the γ isoform while the α/β isoforms unaffected. These observations suggest that an energetic signal may trigger phosphorylation of the p38γ isoform and also may explain how contractions differentially activate signaling pathways. J. Cell. Biochem. 114: 1445–1455, 2013. © 2013 Wiley Periodicals, Inc.     

Quinolines as a novel structural class of potent and selective PDE4 inhibitors. Optimisation for inhaled administration

Crystallography driven optimisation of a lead derived from similarity searching of the GSK compound collection resulted in the discovery of quinoline-3-carboxamides as highly potent and selective inhibitors of phosphodiesterase 4B. This series has been optimized to GSK256066, a potent PDE4B inhibitor which also inhibits LPS induced production of TNF-α from isolated human peripheral blood mononuclear cells with a pIC₅₀ of 11.1. GSK256066 also has a suitable profile for inhaled dosing.     

Plasma and lipoprotein levels of tea catechins following repeated tea consumption

Epidemiological studies suggest that antioxidant flavonoids in tea may reduce the risk of cardiovascular disease, possibly via protection of low-density lipoproteins (LDL) against oxidation. However, the extent of absorption of tea flavonoids and their accumulation in LDL during regular consumption of tea is not clear. Therefore we investigated plasma and lipoprotein levels of catechins during tea consumption and the impact on LDL oxidizability ex vivo. Eighteen healthy adults consumed, in an incomplete balanced cross-over design, green tea, black tea, black tea with milk or water, one cup every 2 hr (eight cups/day) for three days. Blood samples were obtained in the mornings and evenings of each day. Plasma total catechin concentration was determined in all blood samples, and the distribution of catechins among lipoproteins was determined at the end of the third day (t = 60 hr). The resistance of LDL to copper-induced oxidation ex vivo was assessed before tea consumption and at t = 60 hr. Repeated tea consumption during the day rapidly increased plasma total catechin levels whereas they declined overnight when no tea was consumed. There was a gradual increase in plasma levels in the mornings (respectively, 0.08 microM vs. 0.20 microM on first and last day of black tea consumption) and evenings (respectively, 0.29 microM vs. 0.34 microM on first and last day of black tea consumption). Green tea catechins were mainly found in the protein-rich fraction of plasma (60%) and in high-density lipoproteins (23%). Although present in LDL, the concentration of catechins in LDL was not sufficient to enhance the resistance of LDL to oxidation ex vivo. Addition of milk to black tea did not affect any of the parameters measured. In conclusion, the present study shows that catechin levels in blood rapidly increase upon repeated tea consumption. The accumulation of catechins in LDL particles is not sufficient to improve the intrinsic resistance of LDL to oxidation ex vivo.