Characterisation of thymol effects on RDL receptors from the bee parasite Varroa destructor.

A major contributor to bee colony decline is infestation with its most devastating pest, the mite Varroa destructor. To control these mites, thymol is often used, although how it achieves this is not understood. One well-documented action of thymol is to modulate GABA-activated ion channels, which includes insect RDL receptors, a known insecticidal target. Here we have cloned two Varroa RDL subunits, one of which is similar to the canonical RDL subunit, while the other has some differences in M4, and, to a lesser extent, M2 and its binding site loops. Expression of this unusual RDL receptor in Xenopus oocytes reveals GABA-activated receptors, with an EC50 of 56 µM. In contrast to canonical RDL receptors, thymol does not enhance GABA-elicited responses in this receptor, and concentration response curves reveal a decrease in GABA Imax in its presence; this decrease is not seen when similar data are obtained from Apis RDL receptors. We conclude that an M2 T6'M substitution is primarily responsible for the different thymol effects, and suggest that understanding how and where thymol acts could assist in the design of novel bee-friendly miticides.

Cysteine modification reveals which subunits form the ligand binding site in human heteromeric 5-HT3AB receptors.

The ligand binding site of Cys-loop receptors is formed by residues on the principal (+) and complementary (-) faces of adjacent subunits, but the subunits that constitute the binding pocket in many heteromeric receptors are not yet clear. To probe the subunits involved in ligand binding in heteromeric human 5-HT(3)AB receptors, we made cysteine substitutions to the + and - faces of A and B subunits, and measured their functional consequences in receptors expressed in Xenopus oocytes. All A subunit mutations altered or eliminated function. The same pattern of changes was seen at homomeric and heteromeric receptors containing cysteine substitutions at A(R92) (- face), A(L126)(+), A(N128)(+), A(I139)(-), A(Q151)(-) and A(T181)(+), and these receptors displayed further changes when the sulphydryl modifying reagent methanethiosulfonate-ethylammonium (MTSEA) was applied. Modifications of A(R92C)(-)- and A(T181C)(+)-containing receptors were protected by the presence of agonist (5-HT) or antagonist (d-tubocurarine). In contrast modifications of the equivalent B subunit residues did not alter heteromeric receptor function. In addition a double mutant, A(S206C)(-)(/E229C)(+), only responded to 5-HT following DTT treatment in both homomeric and heteromeric receptors, indicating receptor function was inhibited by a disulphide bond between an A+ and an A- interface in both receptor types. Our results are consistent with binding to an A+A- interface at both homomeric and heteromeric human 5-HT(3) receptors, and explain why the competitive pharmacologies of these two receptors are identical.

Burden of migraine. A review of its socioeconomic impact.

According to population-based epidemiological studies using International Headache Society diagnostic criteria, the prevalence of migraine in developed countries ranges from 8 to 14%. These prevalence figures confirm the widespread nature of the disorder. Moreover, as migraine is a chronic episodic disorder that predominantly affects people during their working lives (between the ages of 25 and 55 years), indirect costs associated with reduced productivity represent a substantial proportion of the total cost of migraine. The results of health-related quality-of-life studies demonstrate that migraine has a considerable impact on functional capacity, resulting in disrupted work and social activities. Many migraineurs, however, do not seek medical attention, have not been accurately diagnosed by a physician or do not use prescription medication. Therefore, the direct costs of treatment for migraine are relatively small compared with the indirect costs. Migraine is an important chronic illness that has a major impact on the working sector of a population. The overall cost attributable to migraine is unknown, but it is now established that the indirect costs of migraine outweigh the direct costs and therefore represent an obvious target for healthcare intervention aimed at reducing the impact of this chronic condition.

A multinational investigation of the impact of subcutaneous sumatriptan. IV: Patient satisfaction.

This report describes the patient satisfaction results from a prospective, sequential, multinational study. The study was conducted to concurrently evaluate the effects of sumatriptan, compared with customary therapy, on clinical parameters, health-related quality of life, productivity and patient satisfaction in adult patients with moderate to severe migraine. Patients treated migraine attacks for 12 weeks with their customary therapy, followed by 24 weeks' treatment with subcutaneous sumatriptan 6 mg. A questionnaire was conducted at the end of each study phase, or retrospectively at the end of the study, to assess patient satisfaction with customary therapy and sumatriptan. Sumatriptan was considered by most patients (67 to 85%) to be dependable and fast-acting, and to have a long duration of effect, allowing a quick return to normal activities. By comparison, 15 to 32% of patients considered that their customary therapy possessed the same attributes. However, customary therapy was considered to be easy/very easy to use by 82% of patients compared with 62% for subcutaneous sumatriptan. 89% of patients indicated that they would use sumatriptan again in the future. This study demonstrates that treatment of migraine attacks with subcutaneous sumatriptan for 24 weeks is associated with greater patient satisfaction as regards specific drug attributes than customary therapy.

A multinational investigation of the impact of subcutaneous sumatriptan. I: Design, methods and clinical findings.

This report describes the design, methods and clinical results of a prospective sequential multinational (5 countries) study conducted to evaluate the effects of subcutaneous sumatriptan on health-related quality of life, workplace productivity, clinical parameters and patient satisfaction. Adult patients with moderate to severe migraine initially received customary therapy for migraine episodes for 12 weeks, followed by 24 weeks' treatment with self-administered subcutaneous sumatriptan 6 mg. Demographic, baseline, health-related quality of life and patient satisfaction rating data were collected during visits to the clinic. Data relating to migraine symptoms, migraine therapy, work productivity and non-work activity time were collected on diary cards filled out by the patients. 749 patients were recruited to the study and 637 received at least 1 dose of sumatriptan. Overall, 75.5% of migraines were successfully treated within 2 hours with sumatriptan compared with 31.9% with customary therapy; 36% of patients reported complete relief at 2 hours with sumatriptan treatment compared with 1% of patients receiving customary therapy. 69% of patients successfully treated 70% of their migraines with sumatriptan within 2 hours, compared with 12% of patients with customary therapy. No serious adverse events were reported; 50% of patients reported an adverse event during the 12-week customary therapy phase and 89% of patients during the 24-week sumatriptan phase. These clinical results, which are consistent with those reported in randomised blinded studies of subcutaneous sumatriptan, suggest that relief of migraine symptoms occurs more often, and in less time, in patients receiving subcutaneous sumatriptan rather than customary therapy as their primary medication.

Diet physical form, fatty acid chain length, and emulsification alter fat utilization and growth of newly weaned pigs.

An experiment was conducted to examine the interplay of diet physical form (liquid vs. dry), fatty acid chain length [medium- (MCT) vs. long-chain triglyceride (LCT)], and emulsification as determinants of fat utilization and growth of newly weaned pigs. Ninety-six pigs were weaned at 20.0 ± 0.3 d of age (6.80 ± 0.04 kg) and fed ad libitum 1 of 8 diets for 14 d according to a 2(3) factorial arrangement of treatments with 6 pens per diet and 2 pigs per pen. The MCT contained primarily C8:0 and C10:0 fatty acids, whereas the LCT mainly contained C16:0, C18:0, C18:1, and C18:2. Diet physical form greatly impacted piglet growth (P < 0.001), with liquid-fed pigs (486 g/d) growing faster than dry-fed pigs (332 g/d) by 46%. Pigs fed LCT grew 22% faster (P = 0.01) than MCT-fed pigs; however, effects of emulsifier were not detected (P > 0.1). Furthermore, feed intake and G:F were 15% and 29% greater for liquid-fed pigs, and intake also was 21% greater for pigs fed LCT (P = 0.01). Diet physical form had no effect on apparent ileal fatty acid digestibility, but as expected, digestibility was greater (P < 0.001) for the MCT than the LCT diet (98.5% vs. 93.4%). Emulsification improved digestibility of most fatty acids in pigs fed LCT but not MCT (interaction, P < 0.01). Both jejunal and ileal villi height increased from 7 to 14 d postweaning (P < 0.01). Liquid-fed pigs had greater jejunal crypt depth (P < 0.05) compared with pigs fed the dry diet; however, ileal morphology was not affected by diet physical form, fat chain length, or emulsification. Plasma ketone body concentrations were 6-fold greater in pigs fed MCT than LCT, and the difference was greater in pigs fed dry diets (interaction, P = 0.01). The bile salt concentration in jejunal digesta was 2.2-fold greater in pigs fed LCT than in pigs fed MCT (P < 0.001). Collectively, we conclude that feeding liquid diets containing emulsified LCT can improve fat utilization and markedly accentuate feed intake, growth, and G:F of weanling pigs.

Use of Saccharomyces cerevisiae fermentation product on growth performance and microbiota of weaned pigs during Salmonella infection.

Anaerobically fermented yeast products are a rich source of nutritional metabolites, mannanoligosaccharides, and ß-glucans that may optimize gut health and immunity, which can translate into better growth performance and a reduced risk of foodborne pathogens. The objective of this study was to quantify the effects of Saccharomyces cerevisiae fermentation product (Diamond V Original XPC) inclusion in nursery diets on pig performance and gastrointestinal microbial ecology before, during, and after an oral challenge with Salmonella. Pigs (n = 40) were weaned at 21 d of age, blocked by BW, and assigned in a 2 × 2 factorial arrangement consisting of diet (control or 0.2% XPC) and inoculation (sterile broth or Salmonella). Pigs were fed a 3-phase nursery diet (0 to 7 d, 7 to 21 d, and 21 to 35 d) with ad libitum access to water and feed. On d 14, pigs were orally inoculated with 10(9) cfu of Salmonella enterica serovar Typhimurium DT104 or sterile broth. During d 17 to 20, all pigs were treated with a 5 mg/kg of BW intramuscular injection of ceftiofur-HCl. Growth performance and alterations in the gastrointestinal microbial ecology were measured during preinoculation (PRE; 0 to 14 d), sick (SCK; 14 to 21 d), and postinoculation (POST; 21 to 35 d). Body weight and ADG were measured weekly. Rectal temperature (RT) was measured weekly during PRE and POST, and every 12 h during SCK. Diet had no effect on BW, ADG, or RT during any period (P = 0.12 to 0.95). Inclusion of XPC tended (P < 0.10) to increase Salmonella shedding in feces during SCK. Consumption of XPC altered the composition of the gastrointestinal microbial community, resulting in increased (P < 0.05) populations of Bacteroidetes and Lactobacillus after Salmonella infection. Pigs inoculated with Salmonella had decreased ADG and BW, and increased RT during SCK (P < 0.001). Furthermore, fecal Salmonella cfu (log(10)) was modestly correlated (P = 0.002) with BW (r = -0.22), ADFI (r = -0.27), ADG (r = -0.36), G:F (r = -0.18), and RT (r = 0.52) during SCK. After antibiotic administration, all Salmonella-infected pigs stopped shedding. During POST, an interaction between diet and inoculation (P = 0.009) on ADG indicated that pigs infected with Salmonella grew better when eating XPC than the control diet. The addition of XPC to the diets of weanling pigs resulted in greater compensatory BW gains after infection with Salmonella than in pigs fed conventional nursery diets. This increase in BW gain is likely associated with an increase in beneficial bacteria within the gastrointestinal tract.

Evaluation of the total peroxyl radical-scavenging capacity of flavonoids: structure-activity relationships.

The antioxidant activity of a series of flavonoids against peroxyl radicals generated from thermal homolysis of 2, 2'-azobis-amidinopropane was determined by the Total Oxyradical Scavenging Capacity (TOSC) assay. Seven flavonoids with hydroxy and/or methoxy substitution were analyzed and compared to the water-soluble vitamin E analogue Trolox. The most active compound was the flavonol quercetin, followed by its 3-glycoside derivative rutin; these were 7 and 5 times, respectively, better scavengers of peroxyl radical than Trolox. Among the flavones with both hydroxy and methoxy substitution, the most active against peroxyl radicals was the 5,6,4'-trihydroxy-7,8,3'-trimethoxyflavone (thymonin), with a TOSC value 1.5 times greater than that of Trolox. The activity of the remaining flavones was in the following relative order: 5, 4'-dihydroxy-6,7,8,3'-tetramethoxyflavone > 5-hydroxy-3,6,7,3', 4'-pentamethoxyflavone (artemetin) > 5,4'-dihydroxy-3,6, 7-trimethoxyflavone > 5,6,7,8,2',3',4',5'-octamethoxyflavone (agehoustin A). The data suggest a potential role for dietary intake of flavonoid-containing foods in lowering the risk of certain pathophysiologies that have been associated with free-radical-mediated events.

Association of prokaryotes with symptomatic appearance of withering syndrome in black abalone Haliotis cracherodii.

Withering syndrome (WS) is an epizootic fatal wasting disease that is devastating California Channel Island populations of black abalone Haliotis cracherodii. Our studies suggest a strong pathogen-disease association. The pathogen is an intracellular prokaryote that infects epithelial cells lining the gut and enzyme secreting cells of the digestive diverticula. It multiplies by binary fission in round to oval, basophilic, membrane-bound colonies teeming in the cytoplasm. Infection of the digestive diverticula is accompanied by a complete loss of digestive enzyme granules and metaplasia of enzyme secretory cells to a morphology similar to epithelium lining the gut. Extensive infection of digestive diverticular cells and the resultant deficiency in digestive enzymes correlates to the degree of pedal muscle atrophy and the severity of signs associated with WS. Electron microscopically the intracellular pathogen is a rod-shaped, ribosome-rich, gram-negative, prokaryote with a trilaminar cell wall consistent with the order Rickettsiales. Microbiological and protozoological methods produced no patterns that implicated other types of microbes. Chemical analysis of tissue from animals from a population with WS did not support an association between WS and environmental pollutant exposure to polycyclic aromatic hydrocarbons, polychlorinated biphenyls, or chlorinated pesticides.