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Biomedical subjects

S Higgs

Publications and source records attributed to S Higgs.

At least 19 recordsLinked to original sources

Differential responsiveness to caffeine and perceived effects of caffeine in moderate and high regular caffeine consumers.

RATIONALE: Individual differences in responsiveness to caffeine occur even within a caffeine-consuming population, but the factors that mediate differential responsiveness remain unclear. OBJECTIVES: To compare caffeine's effects on performance and mood in a group of high vs moderate consumers of caffeine and to examine the potential role of subjective awareness of the effects of caffeine in mediating any differential responsiveness. MATERIALS AND METHODS: Two groups of regular caffeine consumers (<200 mg/day and >200 mg/day) attended two sessions at which mood and cognitive functions were measured before and 30 min after consumption of 400-mg caffeine or placebo in a capsule. Cognitive tests included visual information processing, match-to-sample visual search (MTS) and simple and choice reaction times. Post-session questionnaires asked participants to describe any perceived effect of capsule consumption. RESULTS: High consumers, but not moderate consumers, demonstrated significantly faster simple and choice reaction times after caffeine relative to placebo. These effects were not attributable to obvious group differences in withdrawal or tolerance because there were no group differences in baseline mood or in reports of negative affect after caffeine. Instead, the high consumers were more likely to report experiencing positive effects of caffeine, whereas the moderate consumers were more likely to report no effect. CONCLUSIONS: The sensitivity of caffeine consumers to the mood- and performance-enhancing effects of caffeine is related to their levels of habitual intake. High caffeine consumers are more likely than moderate consumers to perceive broadly positive effects of caffeine, and this may contribute to their levels of use.

Adult↗

Differential effects of two cannabinoid receptor agonists on progressive ratio responding for food and free-feeding in rats.

The cannabinoid receptor agonists delta9-tetrahydrocannabinol (delta9-THC) and HU-210 were compared in terms of their effects on: (1) progressive ratio (PR) responding for food, and (2) free food intake. In the first experiment, food-deprived Wistar rats were trained on a time-constrained (60 min) PR-5 schedule for food reinforcement, in which the response requirement incremented by five lever presses for each successive reinforcer. One group of rats received vehicle, 0.5, 1 or 3 mg/kg delta9-THC (i.p.), and three other groups received HU-210 (i.p.) at three different dose ranges, spanning 0.001-0.1 mg/kg. In the second experiment, the effects of the two drugs on free food intake were tested in a separate group of non-deprived rats. For PR responding, delta9-THC significantly increased the break point (final ratio completed) and the total number of lever presses emitted. The same drug also significantly increased free food intake. However, the effects of HU-210 were quite different: it did not alter PR responding at any dose; instead, its only significant effect was to reduce free food intake at 0.06 mg/kg. These data suggest that increased motivation to obtain food might underlie the hyperphagic effects of delta9-THC. However, the synthetic agonist HU-210 has different effects: it only acts to reduce feeding behaviour, an outcome that probably reflects non-specific behavioural disruption. These findings suggest important differences between the two CB1 receptor agonists in terms of their pharmacological effects.

Analysis of Variance↗

Modulation of Anopheles gambiae gene expression in response to o'nyong-nyong virus infection.

To determine if gene expression of An. gambiae is modulated in response to o'nyong-nyong virus (ONNV) infection, we utilized cDNA microarrays including about 20 000 cDNAs. Gene expression levels of ONNV-infected female mosquitoes were compared to that of the uninfected control females harvested at 14 days postinfection. In response to ONNV infection, expression levels of 18 genes were significantly modulated, being at least two-fold up- or down-regulated. Quantitative real-time PCR analysis (qRT-PCR) further substantiated the differential expression of six of these genes in response to ONNV infection. These genes have similarity to a putative heat shock protein 70, DAN4, agglutinin attachment subunit, elongation factor 1 alpha and ribosomal protein L35. One gene, with sequence similarity to mitochondrial ribosomal protein L7, was down-regulated in infected mosquitoes. The expression levels and annotation of the differentially expressed genes are discussed in the context of host/virus interaction including host translation/replication factors, and intracellular transport pathways.

Animals↗

Factors associated with reduced FEV1 in adult patients with cystic fibrosis in a relatively affluent area.

Data collected on adult cystic fibrosis (CF) patients and entered onto the CF database in the south and west regions of England were analysed for the year 2001. FEV1 was taken as a marker of lung disease severity. Data on 371 patients (158 female) mean age 24.7 years (range 16.0-48.9) were assessed. FEV1 was reduced in CF patients infected with Pseudomonas aeruginosa (Pa) (mean 62.6% predicted) compared with those without (mean FEV1 77.8%, P < 0.00001). The reduction was noted irrespective of age group. FEV1 progressively reduced with the increasing need for high-intensity treatment (P < 0.00001) and with the diagnosis of diabetes mellitus (P = 0.03). FEV1 correlated with body mass index (BMI) (r = + 0.42, P < 0.0001). Genetic profile and poverty and deprivation score did not affect the value of FEV1. Chronic infection with Pa mainly in young adults, treatment intensity, diabetes mellitus and reduced body mass index are associated with reduced FEV1 in adult patients with cystic fibrosis in the south and west regions of England. In this relatively affluent area, scores of poverty and deprivation were not associated with the decline in lung function tests.

Adolescent↗

The use of a double subgenomic Sindbis virus expression system to study mosquito gene function: effects of antisense nucleotide number and duration of viral infection on gene silencing efficiency.

Recently we established a simple, effective antisense strategy using a double subgenomic Sindbis (dsSIN) virus expression system to study gene function in mosquitoes. In this study, we further elucidate the effects of antisense nucleotide number and duration of viral infection on mosquito gene silencing efficiency by the dsSIN virus expression system. Over 15 days post virus infection, the degree of parasite melanization was progressively reduced by more than 95%, 75% and 55% in the mosquito Armigeres subalbatus transduced with 600, 147 or 36 bases antisense RNA, targeted to the highly conserved copper binding region of the Ar. subalbatus prophenoloxidase I gene (As-pro-POI), respectively. As the duration of viral infection increased from day 3-15, the degree of parasite melanization progressively decreased in all mosquitoes transduced with antisense RNA, irrespective of the lengths of antisense RNA. Progressive loss of parasite melanization function was found to correlate with down regulation of As-pro-PO expression at both the mRNA and protein activity levels, and reductions in virus titres in mosquitoes transduced with antisense RNA. A small pro-PO RNA (c. twenty-five nucleotides) was identified in mosquitoes transduced with antisense RNA. These data suggest that As-pro-POI gene expression is knocked down by degrading the As-pro-POI mRNA through the RNAi pathway. In conclusion, our study demonstrates that even a short antisense RNA (thirty-six bases) can cause silencing of the As-pro-POI gene, and the effects of endogenous gene silencing by dsSIN expression system on mosquito gene functions can be accumulative.

Animals↗

Infection patterns of o'nyong nyong virus in the malaria-transmitting mosquito, Anopheles gambiae.

Arthropod-borne alphaviruses transmitted by mosquitoes almost exclusively use culicines; however, the alphavirus o'nyong-nyong (ONNV) has the unusual characteristic of being transmitted primarily by anopheline mosquitoes. This unusual attribute makes ONNV a valuable tool in the characterization of mosquito determinants of infection as well as a useful expression system in Anopheles species. We developed a series of recombinant alphaviruses, based upon the genome of ONNV, designed for the expression of heterologous genes. The backbone genome is a full-length infectious cDNA clone of ONNV from which wild-type virus can be rescued. Additional constructs are variants of the primary clone and contain the complete genome plus a duplicated subgenomic promoter element with a multiple cloning site for insertion of heterologous genes. We inserted a green fluorescent protein (GFP) gene downstream of this promoter and used it to characterize infection and dissemination patterns of ONNV within An. gambiae mosquitoes. These experiments allowed us to identify atypical sites of initial infection and dissemination patterns in this mosquito species not frequently observed in comparable culicine infections. The utility of these ONNVs for studies in anopheline mosquitoes includes the potential for identification of vector infection determinants and to serve as tools for antimalaria studies. Viruses that can express a heterologous gene in a vector and rapidly and efficiently infect numerous tissues in An. gambiae mosquitoes will be a valuable asset in parasite-mosquito interaction and interference research.

Alphavirus↗

Differential modulation of murine host immune response by salivary gland extracts from the mosquitoes Aedes aegypti and Culex quinquefasciatus.

Mosquitoes (Diptera: Culicidae) are major vectors of numerous infectious agents. Compounds in mosquito saliva not only facilitate blood-feeding, but may also have an impact upon the immune system of vertebrate hosts. Consequently, the exposure to mosquito saliva may influence pathogen transmission, establishment and disease development. Using two medically important vector mosquitoes, Aedes aegypti (L.) and Culex quinquefasciatus Say, we examined the effects of mosquito saliva on immune cells of host mice. After antigen-specific or non-specific stimulation, murine splenocyte proliferation and production of both Th1 and Th2 cytokines were significantly reduced in the presence of salivary gland extract (SGE) from Ae. aegypti, but not SGE from Cx. quinquefasciatus. T cell populations were highly susceptible to this suppression, showing increased mortality and reduced division rates - judged by flow cytometric analyses. Evidently these two culicine mosquitoes differ in their host immunomodulatory activities.

Aedes↗

The effects of 7-OH-DPAT, quinpirole and raclopride on licking for sucrose solutions in the non-deprived rat.

Pharmacological manipulations that alter dopamine (DA) at DA receptor subtypes produce reductions in feeding behaviour. What remains uncertain is the exact way in which these reductions in feeding are achieved as a consequence of differing drug actions at separate receptor subtypes. In this study our aim was to compare the anorectic effects of the preferential D3/D2 agonists 7-hydroxy-2-(di-n-propylamino)tetralin (7-OH-DPAT) and quinpirole and the non-selective D2/D3 antagonist raclopride on the microstructure of licking responses in non-deprived rats. In a 20-min test, trained adult, male hooded rats had access to one of three solutions: 1%, 3% or 10% sucrose. 7-OH-DPAT (0.1-1.0 mg/kg, i.p.), quinpirole (0.03-0.3 mg/kg, s.c.), raclopride (0.03-0.3 mg/kg, i.p.) or vehicle were injected 20 min prior to the start of the licking test. A lickometer recorded the timing of each lick, from which the microstructural parameters of bout frequency and bout duration were also computed. All compounds reduced the mean bout duration, while 7-OH-DPAT and raclopride also brought about a compensatory increase in bout number. Analysis of the licking rates over the test session showed that 7-OH-DPAT, quinpirole and raclopride decreased the initial rate, without affecting the rate of decline of licking. Changes in licking microstructure (i.e. initial rate of licking and mean bout duration) after the administration of 7-OH-DPAT, quinpirole and raclopride, are consistent with an action of these dopaminergic compounds to reduce palatability.

Animals↗

Effect of mosquito salivary gland treatment on vesicular stomatitis New Jersey virus replication and interferon alpha/beta expression in vitro.

The sensitivity of vesicular stomatitis (VS) viruses to interferon (IFN)-mediated antiviral effects has been well documented. Previous studies in our laboratory have shown the ability of mosquito saliva to enhance vesicular stomatitis New Jersey (VSNJ) virus infection in mice. To investigate the effect of mosquito saliva on virus replication and IFN alpha/beta expression, virus titers were analyzed at various time points after infection in cells that were treated with mosquito salivary gland homogenate (SGH). Salivary gland treatment of mouse fibroblast cells (L929) resulted in a significant increase in virus growth kinetics compared with untreated controls. In contrast, Vero cells, which are deficient in the IFN alpha/beta response, did not yield increased viral titers in the time points examined. Treatment of L929 cells with an IFN alpha/beta neutralizing antibody also slightly increased virus yield. Ribonuclease protection assays revealed that induction of IFN alpha2 expression was reduced in L929 cells treated with SGH. Modulation of IFN alpha/beta by mosquito saliva may be a critical determinant of the transmission and pathogenesis of VSNJ virus.

Animals↗

Molecular characterization of California serogroup viruses isolated in Russia.

Nucleotide sequencing was used to characterize unidentified California (CAL) serogroup virus isolates from Russia. These viruses were isolated from mosquitoes and humans during epidemiologic investigations on the role of CAL serogroup viruses in the increased incidence of arboviral encephalitis in Russia. Most of the isolates were identified serologically as snowshoe hare (SSH), Inkoo (INK), and Tahyna (TAH) viruses, but some of the isolates were difficult to classify serologically, suggesting that they could be reassortant viruses. There is evidence that at least 2 of these viruses are not reassortant viruses. Sequence analysis revealed that the Russian viruses differ from other Eurasian and North American CAL serogroup viruses in all of the segments analyzed. They are most closely related to SSH virus. Whether they differ sufficiently to be considered a new group of SSH-like viruses remains to be determined.

Animals↗

Characterization of an endogenous gene expressed in Aedes aegypti using an orally infectious recombinant Sindbis virus.

Sindbis virus expression vectors have been used successfully to express and silence genes of interest in vivo in several mosquito species, including Aedes aegypti, Ae. albopictus, Ae. triseriatus,Culex pipiens, Armigeres subalbatus and Anopheles gambiae. Here we describe the expression of an endogenous gene, defensin, in Ae. aegypti using the orally infectious Sindbis virus, MRE/3'2J expression vector. We optimized conditions to infect mosquito larvae per os using C6/36Ae. albopictus cells infected with the recombinant virus to maximize virus infection and expression of defensin. Infection with the parental Sindbis virus (MRE/3'2J) did not induce defensin expression. Mosquito larvae infected by ingestion of recombinant Sindbis virus-infected C6/36 cells expressed defensin when they emerged as adults. Defensin expression was observed by western analysis or indirect fluorescent assay in all developmental stages of mosquitoes infected with MRE/3'2J virus that contained the defensin insert. The multiplicity of infection of C6/36 cells and the quantity of infected cells consumed by larvae played an important role in defensin expression. Parental viruses, missing the defensin insert, and/or other defective interfering virus may have contributed to these observations.

Aedes↗

Effects of dietary restraint on flavour-flavour learning.

Flavour preference learning in 21 restrained and 21 unrestrained females was explored using an evaluative conditioning paradigm. Each participant was exposed to an adapted version of the procedure used by Johnsrude et al., (1999, Learning & Motivation30, 250-264). During conditioning, participants sampled 10 instances each of three novel flavours presented in a semi-randomized order. After sampling a flavour they were instructed to eat a sweet according to three different flavour-reinforcement contingencies. One flavour was accompanied by a sweet on 90% of trials and was presented alone on 10% of the trials, while the remaining flavours were rewarded at ratios 50% : 50% and 10% : 90%, respectively. The conditioning phase occurred in conjunction with a counting task requiring continuous use of working memory, and was immediately followed by the participants making hedonic ratings of each flavour. Very few participants showed awareness of the purpose of the experiment or the specific reward contingencies. Despite this, the ratings given by the unrestrained eaters were highly correlated with the reward ratio experienced during conditioning. In contrast, restrained eaters exhibited no evidence for evaluative learning. These findings may explain the equivocal nature of results from previous studies of positive flavour-flavour learning (FFL) and may offer a novel theoretical context within which to study dietary restraint.

Adult↗

The effect of cytotoxic lesions of the hippocampus on recognition memory in the rat: effects of stimulus size.

Rats with excitotoxic hippocampal lesions were trained on delayed nonmatching-to-sample (DNMS) with small goal boxes, containing complex objects, presented on a pseudo trial-unique schedule. A series of experiments then tested performance on repeated presentation of either the small object or large empty goal boxes. All rats acquired the nonmatching rule, but hippocampal-lesioned rats performed less well than controls on choice accuracy for the final 2 blocks of acquisition. In the study's main phase, the lesions impaired choice accuracy when the large empty boxes were used as stimuli. This deficit was ameliorated when the rats were tested with the small object boxes, although the performance of the hippocampal-lesioned rats was still below that of controls. These results extend previous reports of box size-dependent effects of hippocampal aspiration lesions on DNMS and suggest that selective damage to the hippocampus, not neuronal loss in adjacent structures or fiber tracts, is critical for the effect.

Animals↗

Effect of prophenoloxidase expression knockout on the melanization of microfilariae in the mosquito Armigeres subalbatus.

Melanization is an effective defence reaction used by mosquito hosts to kill malarial and filarial worm parasites. Although phenoloxidase (PO) has long been considered to be the key enzyme in the biosynthesis of melanotic material in insects, there is no direct evidence verifying its role in parasite melanization. To elucidate the role of PO in the melanization of microfilariae (mf) by mosquitoes, a double subgenomic Sindbis (dsSIN) recombinant virus was used to transduce Armigeres subalbatus mosquitoes with a 600 base antisense RNA targeted to the highly conserved copper-binding region of an Ar. subalbatus PO gene. Compared with controls, haemolymph PO activity in mosquitoes transduced with antisense RNA was significantly reduced. When these mosquitoes were challenged with Dirofilaria immitis mf, the melanization of mf was almost completely inhibited. These data verify that PO is an essential component of the biochemical pathway required for the melanization of parasites, and that the dsSIN expression system represents a useful tool in the functional analysis of endogenous gene expression in mosquitoes.

Animals↗

The effect of the dopamine D2 receptor antagonist raclopride on the pattern of licking microstructure induced by midazolam in the rat.

The role of dopamine in the effects of midazolam on ingestive behaviour was investigated using microstructural analysis of licking behaviour in the rat. Midazolam (1.8 mg/kg i.p.) was administered alone or in combination with the dopamine D2 receptor antagonist raclopride (0.1 and 0.3 mg/kg i.p.). The effect on licking patterns during 60 s exposure to a range of concentrations of sucrose solution was recorded using an automated lickometer. Midazolam increased the total number of licks via an increase in mean bout duration, an effect consistent with the proposal that these drugs enhance palatability. Midazolam also decreased the intrabout lick rate, probably because of muscle relaxant effects. Pre-treatment with raclopride blocked midazolam-induced increases in mean bout duration, at doses that by themselves were ineffective, but did not reverse the decrease in intrabout lick rate. These data point to the interdependence of benzodiazepine and dopamine substrates in the mediation of palatability.

Animals↗

Population biology of Eimeria (Protozoa: Apicomplexa) in Apodemus sylvaticus: a capture/recapture study.

The first long-term (27 month) survey of single species of Eimeria occurring in a population of Apodemus sylvaticus in the UK showed that Eimeria apionodes, E. hungaryensis and E. uptoni occurred throughout the period whilst E. sp. E. was only found during 4 months. In first-captured animals, overall prevalence of Eimeria spp. was 73%: a figure higher than previously published, but possibly an underestimate. Multiple infections (2 or 3 species) occurred in 34% of first-capture infected animals, but without significant associations. There were no significant differences of prevalence rates between the sexes, nor between adults and juveniles, except for E. uptoni which was more common in juveniles than in adults. E. hungaryensis and E. uptoni probably occurred as near-continuous infections whilst E. apionodes occurred more intermittently but with no long-term total immunity. Monthly prevalence data indicated ongoing trends over the 27 months. Prevalences of E. apionodes indicated an annual cycle, lowest in January-May and with a peak in October-November, correlating positively with both the number of animals and the percentage of juveniles in the host population and also with the environmental relative humidity. There was no consistent pattern for E. uptoni and an annual cycle for E. hungaryensis was not apparent.

Age Factors↗

Development of a Sindbis virus expression system that efficiently expresses green fluorescent protein in midguts of Aedes aegypti following per os infection.

A double subgenomic Sindbis (dsSIN) virus, MRE/3'2 J/GFP, was constructed to efficiently express green fluorescent protein (GFP) in the midgut of Aedes aegypti following per os infection. The MRE/3'2 J/GFP RNA genome contained the nonstructural genes and cis-acting sequences of the dsSIN virus, TE/3'2 J/GFP, but had the structural genes of MRE16 SIN virus. MRE/3'2 J/GFP virus, unlike TE/3'2 J/GFP virus, efficiently infected mosquitoes orally. At 1-2 days postinfection, GFP was observed as multiple foci of expression on the lumenal side of the midgut. At 10-12 days postinfection, thirteen of fifteen mosquitoes infected with MRE/3'2 J/GFP virus had high levels of GFP expression in the mosquito midgut. The MRE3'2 J dsSIN expression system should be an important tool for efficient gene expression in Ae. aegypti midguts.

Animals↗

Chemical and gamma-ray mutagenesis of the white gene in Aedes aegypti.

A molecular understanding of an insect gene can be facilitated by analysing the phenotypes of mutants for that gene. Protocols were developed for both chemical and gamma-ray mutagenesis in Aedes aegypti using the white (w) gene as an assay. Wild-type adult males were subjected to varying doses of either ethyl methanesulphonate (0. 1%, 0.5% or 1%) or gamma rays (1500 R or 3000 R), mated to females homozygous for the recessive w mutation, and progeny screened for the w phenotype, indicating non-complementation. The expression of newly induced w alleles was either complete or mosaic. Gamma-ray mutagenesis resulted in high (1.65 or 6.39%, depending on dose) induction of mutant alleles for the w gene, but not for a different gene, red-eye (0.15%). Gamma-ray-induced w alleles did not revert at a reasonable frequency following additional irradiation, suggesting that the high rate of gamma-ray-induced w mutagenesis is not due to a transposon insertion event.

Aedes↗