ALBERT

Notes: Surveys were conducted in three fishing communities in Ghana, where an improved fish smoking technology (the Chorkor smoker) had been introduced previously, to assess the impact of adopting the improved technology on household income, health and nutritional status. Fifty-one fish processors (all women), who had adopted the improved fish processing technology, were selected for the study, in which the participatory rural appraisal (PRA) technique, nutritional survey methods and structured questionnaires were used to obtain both qualitative and quantitative data. The results revealed that there was an improvement in economic activities. This resulted in a positive impact on household income and food consumption patterns of the processors. Improvement in household income resulted from increases in the quality and quantity of output, price per unit output and profits. Improvement in food consumption pattern was mostly in respect of quality and quantity of food intake, but the types of food consumed remained the same. The nutrient intake of the respondents was good on average, with a majority exceeding the recommended daily allowance (RDA) of protein, vitamin B1 and calcium. The average body mass index (BMI) of the processors was also within the ideal range of 20–25. Adoption of the new technology also improved the health status of the respondents. Lessening of eye problems and headaches as a result of reduced exposure to smoke and heat was reported by 52% of the women. However, preschool children were undernourished as a result of poor child feeding practices. In any technology transfer process, apparently unrelated nutritional factors need to be considered.

Notes: This paper explores a consumer complaint programme as a tool for empowering consumers, using the consumer complaint programme of a United States federal agency as a case study. After describing the structure and role of the Federal Reserve System (the central bank of the United States) in handling complaints, the paper follows a complaint through the System, looks at trends in complaints and provides a profile of consumers who complain. The paper concludes with information on the economic impact of complaint resolution and consumer satisfaction with third-party complaint investigation.

Notes: The curriculum that pupils are taught within the classroom environment can either be reinforced or weakened by what goes on in the rest of the school environment. The study focused on pupils aged 11–12 years, to establish if healthy eating theory was put into practice in the school environment. The research encompassed three areas within the school: the pupils who are the recipients of the theory; the teacher who teaches them the importance of food and cooking skills; and the providers of food within the school environment. Three different post-primary schools located within the Coleraine area of Northern Ireland were used.

Notes: This paper examines questions concerning consumer education in relation to consumption and household management. It is based partly on literature studies and partly on a current pilot study, also on studies carried out in the classroom and developmental work in schools and on teacher training courses. The pilot study on consumer education is being carried out in Denmark and is funded by the Danish Ministry of Business Affairs. Another part of the study concerns a qualitative investigation of pupils’ understandings of consumption and its meaning in their lives, but this is not reported here. The key research questions relate to the way in which the young consumer is educated, both formally and informally, and what the possibilities and perspectives are for consumer education. Introductory research is discussed, followed by a presentation and discussion of key issues for consumer education, such as household management, consumption, home economics and education. Finally, three examples are described and discussed which demonstrate how the advocated principles of consumer education and empowerment can be put into practice. These examples are based on developmental work carried out in lower secondary schools and teacher training courses.

Notes: The General Consumer Council for Northern Ireland (GCCNI) is one of the four Consumer Councils operating within the United Kingdom (the others are the Welsh, the Scottish and the National Consumer Council, which covers England and links with the other Consumer Councils). The GCCNI has a long standing interest in consumer education, and has produced a quantity of well-regarded consumer education material aimed at both schools and adults over recent years. It has also organised regular consumer education events for schools and conferences on this subject., However, the GCCNI is aware that many people in Northern Ireland suffer difficulties in relation to consumer issues and is looking for a clear policy on dealing with those difficulties. They contracted researchers at Queen's University, Belfast to investigate the extent of the problem, with the aim of identifying the key aspects of a new consumer education and skills development strategy and the gaps and deficiencies in current provision that need to be addressed. While the GCCNI acknowledges that these problems are not exclusive to Northern Ireland, and that the nature of the problems is relatively well-known, they felt that conditions in Northern Ireland were unique and deserving of a specific and targeted solution. Policy and decision makers in other countries will find the Northern Ireland experience of interest, both from the way in which the problem was approached and the solution which evolved.

Notes: The purpose of the study is to investigate antecedents and consequences of consumer satisfaction/dissatisfaction (CS/D) with the performance of apparel products at purchase, and after consumption, for male and female South Korean consumers. An experimental research technique with a 2 × 2 × 2 between-subjects factorial design with gender (female, male), expectation for future performance, (high, low) and perception of product consumption performance (good, poor) was used. Both similarities and differences between genders were found. Implications and applications of results are discussed.

Notes: Research on how Chinese consumption values influence Chinese consumer behaviour is rare. First, this paper examines consumers’ attitudes towards the physical and intangible attributes of imported fruit. Then, it identifies consumers’ consumption values and the role of these values in purchasing behaviour. Data were collected through point of sale intercept surveys conducted in Guangzhou, China. Latent consumption values of consumers were identified through factor analysis. K-means clustering revealed four natural groupings of consumers, each group demonstrating different consumption values. The results demonstrated the primary importance of symbolic values and hedonic values in the decision to purchase imported fruit. Such consumption values may derive from the intermingling of Confucian and Western cultural values. Results from this study could help to better understand interrelationships among product attributes, consumption values and cultural values, and could make a significant contribution in developing strategies to market imported fruit in China.

Notes: This paper uses the binary choice model to identify the factors that are significantly influencing the household purchase decisions of seafood products for home consumption in Auckland, New Zealand. It is found that ‘quality’ and ‘cooking easiness’ are the main product attributes that significantly influence households’ choices of seafood in Auckland. Also, the representative household has shown a strong preference for fresh and other alternative seafood products, including processed, smoked and canned, over frozen products. Retail outlets are found to be more attractive to the household purchasing seafood for home consumption. The New Zealand seafood industry may find this baseline study useful as a guide to developing future research structure on the domestic market.

Notes: Samples of mayonnaise were prepared in which the oil content was reduced from 30% to 10% and replaced with the fat replacers, Avicel, N Lite S and Simplesse. The control sample had a significantly higher viscosity (P〈 0.001) and droplet size (P〈 0.01) than the samples containing the replacers. There was a significant (P〈 0.001) correlation (r= 0.97) between viscosity and droplet size. Sensory analysis also showed that the control had significantly higher scores (P〈 0. 001) for the attributes of thickness, meltability, glossiness and oiliness. There was a significant (P〈 0.05) correlation (r= 0.93) between viscosity and sensory thickness. However, there was no significant preference for any of the samples, showing that variations in physical properties and sensory profile attributes do not necessarily affect preference. The use of the replacers reduced the energy values of the mayonnaises by 52% to 55%.

Notes: The long-established association between high salt consumption and hypertension has led to calls for a reduction in salt intake. There is particular concern for the health status of Northern Irish consumers, who already have among the highest rates of coronary heart disease and stroke in the United Kingdom. With these concerns in mind, it is essential to establish the sources and patterns of salt consumption from discretionary and processed food sources before a reduction can be achieved. This study investigated differences in salt consumption patterns among consumers of varying socio-economic status. Research consisted of a consumer questionnaire (n= 360) distributed within Northern Ireland. The results of the study revealed high discretionary salt usage and frequent consumption of processed foods among consumers of lower socio-economic status. General observations from the questionnaire would suggest a need for improved health education for consumers in this group. The objective of health promotion should be to encourage a reduction in table and cooking salt usage and, in particular, in the consumption of highly salted processed foods. Enhanced product labelling is also encouraged to increase consumer awareness of current salt levels in food and of the availability of low-salt foods. In addition, the results of this study advocate further development activity in the production of product formulations with low-salt status.

Notes: Eighty-four primary school children (the majority aged 7–8 years), from four schools, two in England and two in Scotland, were surveyed to identify snacking behaviour and the influences on perception and choice of snack foods. A questionnaire plus short interview with activity tasks was adopted, covering general snacking habits and a look at six specific groups of snacks comprising three of ‘unhealthy’ image and three with intermediate or fully ‘healthy’ image. The results showed that, for this age group, there was an overall preference for sweets, crisps and chocolate and less liking for fruit and cereal-containing snacks. Attitudes to sensory properties showed that ‘taste’ was the main reason for snack choices, usually in spite of a ‘healthy’ image belief for the snacks. Choice was also dependent on parental influence and availability. Gender and relative socio-economic level differences were minimal, but regional variations showed higher snacking frequency for the Scottish group, which also had fewer children preferring fruit. Meal patterns had little effect on snacking, but inclusion of a prepared meal at lunchtime may have reduced snacking frequency.

Notes: The study was conducted to determine the food consumption patterns, dietary quality and health status of expectant women and also to assess their level of awareness of nutritional requirements during pregnancy. Food beliefs, taboos and superstitions held by the women were also investigated. Using the purposive sampling technique, 30 pregnant women attending the Maternal and Child Health Clinic at Dodowa (rural) and University Hospital, Legon (suburban), were selected for the study. A structured questionnaire was used to collect information on the respondents’ nutritional knowledge, beliefs, taboos and superstitions and health status. A food frequency questionnaire and the 24-h dietary recall method were used to obtain information on food consumption patterns and dietary quality. The study revealed that, although the majority of the respondents (83.3%) had some knowledge of nutrition, not all were applying it in their feeding practices because of financial constraints. With regard to consumption patterns, most of the women (83.3%) were having three meals a day, while the rest either ate twice a day or anytime they were hungry. Seventy-three per cent of the respondents also increased their food intake during pregnancy. Foods eaten were based mainly on starchy roots and tubers, cereals and vegetables. Legumes, oilseeds and fruits were often lacking in the main meals of respondents. Although animal products were consumed daily, the quantities taken were very small to provide for adequate protein, especially in the rural area. Various types of food were avoided by some of the expectant mothers for reasons such as nausea, loss of appetite, taboos and superstition. With regard to dietary quality, respondents from the rural area had diets of poorer quality in terms of nutrient intake compared with those from the suburban area. In both communities, iron contents of diets were quite low. Forty-seven per cent of the women interviewed were anaemic, with a higher prevalence of anaemia being observed in the rural community. Common ailments reported by the women were dizziness, headache, waist pains and malaria. Based on the results of the study, it was recommended that nutrition education for pregnant women should be stepped up at antenatal clinics especially in rural areas.

Notes: Everyday consumer transactions have the same potential for unexpected consequence whatever the age of the consumers involved. Young and old alike can find that products and services fail to live up to performance claims and that they are left with problems not easily resolved, or costs that are difficult to recover. While not overlooking consumer heterogeneity – especially on the basis of age – older consumers are arguably distinguishable in terms of the social and financial context in which they make decisions and attempt to redress problems. In 1988, attention was drawn to the need for consumer education to look beyond generic objectives to the specific situation of older people and their transactions. More than a decade later, in an allegedly consumer-oriented society, the issue is revisited here to assess the argument's current relevance. Despite the increased availability of information for decisions and consumer protection, difficulties persist in the way information is presented or accessed. Chameleon-like, old problems become manifest in new unfamiliar ways and invalidate experience. Consumer education today is as important as it was in 1988. Arguably, technological change means that the need for a better understanding of dangers, rights and redress procedures is greater than ever and the needs of older people in increasingly complex private and public sector transaction environments are all the more pressing. However, a fundamental revision of the way we approach the design of products, services and environments is needed to improve prospects for older consumers.

Notes: All too often, we are told via the media that our children are overweight, under-exercised and eating poor diets. This is disconcerting at the best of times but even more so when we, as a country, are reviewing our farming and food production practices. So what is our perception of food and how does this relate to the lives we lead? These questions were put to initial teacher-trainees (ITT) at the University of Gloucestershire. They will be the teachers of tomorrow, playing a significant role in the development and lifestyles of our children. The answers received were varied; both encouraging and giving cause for concern. This paper describes an action research project based around the analysis of those answers in the light of current research into food and health. Throughout the paper, published research findings relating to school children's perception of food and nutrition are compared with the trainee teachers’ views. The comparison draws striking similarities. The implications of this point to the way we teach food and nutrition in schools and on our ITT programme at the University of Gloucestershire. Whilst praising the good practice that already exists, this paper recognises that there is a need to adequately equip trainee teachers, especially at primary level, to educate children towards a more holistic understanding of the food we eat and how it relates to the lifestyles we lead.

Notes: The aim of this study was to investigate consumer perception and sensory quality of functional dairy produce in Northern Ireland. Consumer awareness and perception of functional foods were investigated using a structured questionnaire (n= 93). A descriptive profiling test1 was used to evaluate the sensory characteristics of selected margarines within the functional dairy food sector (n= 4). The research findings indicate that the categorical variables of gender (P〈 0.001), age (P〈 0.01), education level (P〈 0.05) and occupation (P〈 0.05) are significantly related to the level of consumer awareness to the concept of functional food. Hence, consumer awareness of functional foods is a key factor influencing future acceptance of functional produce. This study also suggests that there is a strong need for clinical trials to support the role of functional ingredients in achieving optimal health.

Notes: Pyramid Power is a 1-hour programme designed to teach food choices, serving sizes and nutrition. Participants (19 women and 8 men) were challenged to follow the recommendation in a step-by-step manner. Evaluation, two months after the programme, resulted in two major areas of change. Participants improved their scoring of three food groups (bread-cereal-rice-pasta, vegetable and fruit) and decreased the number of fat–sweet mentions. The programme and challenge provides a way for nutritionists to identify what participants are eating, to track progress and to identify future programme needs.

Notes: The inclusion of reduced-fat foods in the daily diet affords consumers the potential to reduce overall dietary fat. Nevertheless, despite an increased range of these food products throughout the market, there has remained a lack of widespread consumer acceptance. The aim of the study was therefore to understand why consumers choose or reject reduced-fat foods. Ninety consumers were interviewed by means of a qualitative approach within food retail outlets in the United Kingdom. Thematic content analysis indicated that uptake or rejection of these products is influenced by health concerns, ‘goodness of fit’ with individual dietary health strategies, the preferences of different household members, perceived sensory properties and scepticism towards these products. The results imply a need to explore and innovate ‘natural’ fat substitutes and to improve the sensory quality of reduced-fat products. Further research is required to determine how these foods are used within the household and incorporated within the overall diet. Policy must seek to restore consumer trust in reduced-fat food products and the food supply generally.

Notes: Even though the permanent income and relative income hypotheses have been introduced in past research to explain consumer behaviour, no previous attempt was undertaken to integrate them in one model. In this article, the hypotheses were synthesized into a single model. The model was empirically tested using data from the 1996 Canadian Family Expenditure Survey. The results indicate that household consumption behaviour is generally explained by both hypotheses when integrated into one model, contrary to previously treating them as mutually exclusive hypotheses.

Notes: In 1999, Central Government launched an initiative to establish Consumer Support Networks (CSNs) throughout Scotland, England and Wales. The purpose of the networks is to improve access to high quality consumer advice services. Information and advice agencies will join together to provide expert, accurate and timely advice for consumers. By April 2002, 191 networks had been established covering in excess of 99% of the population. Only one local authority area in England has yet to establish a network. This paper reviews the development of consumer information and advice provision in the UK and considers the context for establishing CSNs. It also examines the need for consumer advice, the role of Central Government and the quality framework. Each CSN will conduct a gap analysis and assess the need for consumer advice in the local area. This will enable the network to develop a good understanding of consumers in the area and to take a more strategic approach in planning service developments to meet the identified needs. Consumer Support Networks aim to improve the provision of consumer information, education and advice. The literature suggests that consumers who are well educated regarding their rights are empowered through an increased ability to exercise them. Clearly branded advice services, marketed effectively to the public should result in higher levels of consumer awareness and, consequently, improve access to services. It is also expected that better flows of information between agencies will result in greater protection for consumers. Sharing ‘best practice’ nationwide should stimulate service improvements throughout.

Notes: This paper addresses the following subjects: biotechnology and consumers, concern about risks, consumer acceptance, labelling of foods produced using biotechnology, the legal approach to consumer protection, and consumer protection policies relating to biotechnology products in the European Union, the United States, Turkey and global institutions such as the Convention for Biological Diversity (CBD) and the World Trade Organisation (WTO). It is likely that biotechnology will gain ground much more rapidly in the twenty-first century than in the past. Despite rapid, detailed and precise advances in gene technology, its applications have not been the received with a great deal of consumer enthusiasm. Consumers have approached genetically modified foods with considerable apprehension and opposition. Consumer concerns about bioengineered food products focus on the questions of human health, environmental and social risks and benefits. The most important stages in the process of marketing new foods produced using biotechnology are to demonstrate user need and consumer acceptance. Generally, the technical complexity of biotechnology makes it difficult for consumers to understand details of the product and the specific attributes of biotechnology applications. Scientific uncertainty, the nature of consumer concerns and general reluctance to accept biotechnology products, increase the importance of consumer protection. Legal protection is a very important factor in the solution of new social problems related to technological advances. More specifically, consumer and environmental law support consumer protection related to foods produced with biotechnology. The basic principles of consumer law can be re-formulated as consumer rights. Environmental law is a new phenomenon, but precautionary principles and public participation in decision-making for environmental law are relevant to consumer protection relating to bioengineered food products.

Notes: The four-dimensional nature of the Kaplans’ model of aesthetic response to images was tested on Point-of-Purchase (POP) material. It was found that only two robust dimensions emerged in the analysis, Clarity and Mystery. These however, had a significant influence on the subjects’ rating of the POP’s attractiveness. Path analysis suggested each of these dimensions was independent of the other. The implications of the findings are discussed and possible future areas of research identified.

Notes: Many variables have been proposed as additions to the theory of planned behaviour structure, and evidence exists to support the value of a measure of ethical obligation and self-identity. Furthermore, some research has suggested that ethical obligation may serve as an antecedent to attitude as well as intention. This paper presents findings from a large scale survey that highlights the usefulness of ethical obligation and self-identity in the prediction of intention. Additionally, the role of both these variables in the prediction of attitude is also suggested. This is examined within the ‘ethical’ context of fair trade grocery purchases. Methodological implications for further research are discussed.

Notes: This paper first presents evidence from Spain on the relationship between the labour situation of both spouses within the family and the time dedicated to the care of the children, the elderly and the infirm. Second, it analyses the effect that government benefits for such care would have on the labour market and on household poverty. The results show that while Spanish society has advanced in the incorporation of women into the labour market, most of them still have to assume total responsibility for housework and the care of the children, the elderly and the infirm. Against this background, we find that benefits would decrease female participation in the labour market and the number of hours worked, while they would also contribute towards reducing poverty.

Notes: The Food Standards Agency was set up in April 2000 to protect the health of the public and the interests of consumers in relation to food. Its principal purpose is to put the consumer first. But what does this mean in practice? The Agency is finding ways to help consumer organizations and consumer representatives contribute more effectively to its work. However, consumer organizations cannot hope to represent the full range of consumer perspectives, and so the Agency is keen to reach out to individual consumers using a variety of innovative mechanisms. This article will describe why the Agency was established and how it achieves its main objective of putting the consumer first. It will discuss how the Agency is developing and testing new ways of working to ensure that the consumer perspective is fully accounted for when developing and evaluating policies, including setting up an influential new Consumer Committee. It will go on to highlight a number of recent innovative consumer consultations and structures that have been established to involve consumers in Agency policy making. It will discuss these processes and how the lessons that have been learnt are informing future Agency strategies. The article will then describe how the Agency protects consumers through effective risk communication strategies and how its Annual Consumer Attitudes Survey will help it measure success so that it can continue to set new stand-ards in putting consumers first in relation to food.

Notes: Despite the increased documentation of consumers’ purchases of organic food products, the motivations for such purchases are relatively under-researched. An individual’s choice of food products can be linked clearly to ethical stances, but ethical choices can also vary from individual to individual, from industry to industry and among countries. Consequently, this paper investigates the degree to which ethical beliefs influence Scottish consumer perceptions, beliefs, attitudes and purchasing decisions, with regard to organic dairy products. Consumer purchasing motivations are revealed as being self-interest-centred (i.e. better tasting, safer), rather than altruistic. Therefore, to achieve future market development, organic dairy producers cannot rely upon the minority of hardcore green consumers to sustain growth, but must aim to modify perceptions and attitudes of larger consumer segments by implementing educational marketing campaigns that reinforce the ethical, environmental and societal benefits of organic production.

Notes: Physically disabled women today have to face the inconvenience of shopping for their own as well as their families’ clothes and the inability of finding fashionable clothes that fit any disability. A total of 40 physically disabled women between the ages of 21 years and 60 years were interviewed on the problems they experienced when shopping for fashionable clothes that fit their disabilities. A total of 40 clothing shops were observed to get information on the extent to which the shops cater for the needs of the physically disabled consumer. Results from the interviews showed that the majority of physically disabled female clothing consumers experienced major problems in finding fashionable clothes that fit their disabilities. It was also clear that these women experienced problems with unusable access routes, unsuitable parking lots, display racks that are too high, spaces between racks that are too narrow, inadequate space in fitting rooms and a lack of assistance in shops. The observations were compared with the complaints the women had and it was found that these women had not overestimated their problems.

Notes: Three denim jeans products from the same manufacturer with differentiated pricing and labels (antiqued, sandblasted, and stonewashed) were used to determine the relationship between price and quality. Both qualitative and quantitative procedures were used to analyse the garments. All three jeans were made of twill weave with 3 × 1 repeat. Both structural and performance characteristics were compared using standardized tests. Analysis of variance was used to compare the selected structural and performance characteristics among the three jeans products. Tukey’s HSD tests were used for post hoc multiple comparisons for three jeans. The findings revealed that despite the fact that all three jeans products were from the same store, and that the fabrics used the same weave, degree and direction of twist, the jeans differed significantly for several structural and performance characteristics. Several possibilities for future research were explored.

Notes: 1 The present study attempted to pharmacologically characterize the subtypes of α-adrenoceptors mediating the vasoconstriction in the isolated and perfused canine vesical artery. 2 Noradrenaline (NA) and phenylephrine (PE, an α1-adrenoceptor agonist) induced a dose-dependent vasoconstriction, whereas xylazine (an α2-agonist) did not induce any clear vascular constrictor response. 3 Prazosin at 0.01 μm and rauwolscine at 0.1 μm failed to affect the NA-induced vasoconstriction. Prazosin at 0.1 μm antagonized the vasoconstrictor responses to NA, with pKB value of 7.8. 4 WB 4101 at 0.01–0.1 μm dose-dependently inhibited the responses to NA, with a pKB value of 8.9. The vasoconstrictor responses to NA were not significantly affected by chloroethylclonidine (10–30 μm) or BMY 7378 (0.1 μm). 5 The present results indicate that the canine vesical arteries dominantly contain α1-adrenoceptors but have no α2-adrenoceptors, and the functional subtype of α1-adrenoceptor is characterized as an α1A-adrenoceptor subtype.

Notes: 1 In this study, we have investigated the vasodilator response to acetylcholine under diabetes conditions in isolated renal arteries of rabbits. We have also examined the contribution of endothelium-derived nitric oxide (EDNO) and endothelium-derived hyperpolarizing factor (EDHF) to the endothelium-dependent relaxation caused by acetylcholine in the renal arteries of alloxan-induced diabetic rabbits. 2 Acetylcholine (10−10−10−4 m) produced cumulative concentration–response curve in the renal arteries of both control and diabetic rabbits. The EC50 values and maximal responses to acetylcholine were not significantly different relative to diabetic conditions. In order to isolate the EDHF component of acetylcholine-induced vasodilator response, l-nitro-methyl arginine ester (l-NAME, 10−4 m) and indomethacin (10−6 m) were added to the Krebs' solution throughout the experiment. Under these conditions, acetylcholine induced vasodilatation in the isolated renal arteries from both control and diabetic rabbits. The vasodilator response to acetylcholine was not affected under diabetic conditions. 3 Sodium nitroprusside (SNP)-induced relaxation was increased in the diabetic rabbits compared with the control animals. 4 Tetrabutyl ammonium (TBA, 0.5 mm) produced a significant reduction in acetylcholine-induced vasodilatation in both preparations from control and diabetic animals, consistent with involvement of K+ channels in mediating this response. Glibenclamide (1 μm) attenuated acetylcholine-induced vasodilatation in preparations from control animals only, while iberiotoxin (0.05 μm) significantly reduced the vasodilator response to acetylcholine in preparations from both control and diabetic animals. 5 The role of EDNO in mediating acetylcholine-induced vasodilatation was examined. The vascular preparations were incubated with 20 mm K+-Krebs' solution to inhibit the EDHF contribution to acetylcholine-induced vasodilatation. Under this condition, acetylcholine induced a vasodilator response in both preparations from control and diabetic rats. Pretreatment with l-NAME (10−4 m) attenuated acetylcholine-induced vasodilatation in both preparations, indicating an nitric oxide-mediated vasodilator response. 6 Our results indicated that acetylcholine-induced vasodilatation in the isolated renal arteries of alloxan-induced diabetic rabbits was not affected under diabetic conditions. Acetylcholine-induced vasodilatation is mediated by two vasodilator components; namely, EDHF and EDNO. The contribution of EDHF and EDNO to acetylcholine-induced vasodilatation was not affected under diabetic conditions and there was no indication of endothelial dysfunction associated with diabetes. EDHF component was found to act mainly through high conductance Ca2+-activated K+ channels under normal and diabetic conditions, while the adenosine triphosphate-dependent K+ channels were involved in mediating acetylcholine vasodilator response in the control preparations only.

Notes: 1 It has been suggested that opioids may play an indirect role in the regulation of the peripheral circulation through the control of nitric oxide (NO) release in vascular tissue. The current study was undertaken to investigate the effect of nitric oxide synthase (NOS) blockade on responses to morphine in phenylephrine (PE)- or KCl-precontracted rat aortic rings. 2 Morphine (3 × 10−8−3 × 10−5 m) administration did not cause any significant effect on basal tonus of endothelium-intact or endothelium-denuded preparations. Morphine produced concentration-dependent relaxation responses in endothelium-intact as well as in endothelium-denuded rat aortic rings precontracted by PE or KCl. Removal of endothelium did not significantly alter the relaxation responses to morphine. 3 The relaxant responses to morphine were significantly and partially inhibited by pretreatment of tissues with naloxone (NAL, 3 × 10−5 m) for 5 min. The inhibitory effect of NAL on relaxant responses to morphine in PE- or KCl-precontracted rings did not differ significantly between endothelium-intact and endothelium-denuded preparations. 4 Incubation of endothelium-intact or endothelium-denuded rat aortic rings with NOS inhibitor, Nω-nitro-l-arginine methyl ester (l-NAME, 10−4 m) for 20 min did not cause a significant inhibition on relaxation responses to morphine. 5 These findings confirmed the presence of opiate receptors in rat thoracic aorta, but suggested that mechanisms other than NO release play a role in the relaxant effect of morphine on rat aortic rings.

Notes: Summary 1 It was shown recently that stimulation of cardiac muscarinic M2-receptors revealed an enhanced negative inotropic response in isolated rat left atria after exposure to hypochlorite-induced oxidative stress. This phenomenon was not observed after stimulation of the cardiac A1-receptor, which like the M2-receptor is coupled to Gi-proteins. Since even the contractile response to M3-receptor stimulation was not amplified in the rat portal vein, we hypothesized a M2-receptor specificity of this hypochlorite-induced enhancement. 2 The present study was performed in order to investigate whether the sympathoinhibitory response to presynaptically located M2-receptor stimulation would also be modified after exposure to hypochlorite in the rat tail artery. We applied electrical field stimulation (EFS) in order to mimic sympathetic neurotransmission. 3 EFS increased the vascular tone frequency-dependently (0.3–4 Hz). EFS-induced vasoconstriction could be attenuated by acetylcholine (30 nm−1 μm) in a concentration-dependent manner. Hypochlorite (10 and 100 μm) did not affect the sympathoinhibitory effect of acetylcholine (100 nm). 4 In conclusion, in contrast to cardiac M2-receptors, hypochlorite did not amplify the sympathoinhibitory effects of presynaptic M2-receptors. The different responsiveness between neuronal and cardiac M2-receptors to hypochlorite may be explained by the different G-protein subunits involved in the activation of the underlying signalling cascade.

Notes: 1 This study was aimed at evaluating the effect of rutin and harmaline (1-methyl-7-methoxy-3,4-dihydro-β-carboline) on the development of the surgically induced reflux oesophagitis, on gastric secretion, lipid peroxidation, polymorphonucleocytes (PMNs) accumulation, superoxide and hydroxyl radical production in PMNs, cytokine [interleukin-1β (IL-1β), tumour necrosis factor-α (TNF-α)] production in blood and [Ca2+]i mobilization in PMNs. 2 Rutin and harmaline significantly prevented the development of reflux oesophagitis and gastric secretion. Treatments of oesophagitis rats with rutin and harmaline inhibited lipid peroxidation, and myeloperoxidase (MPO) in the oesophagus in comparison with untreated rats. 3 Superoxide anion and hydrogen peroxide production in 1 μm formylmethionylleucylphenylalanine (fMLP)- or 0.1 μg ml−1N-phorbol 12-myristate 13-acetate (PMA)-activated PMNs was inhibited by rutin and harmaline in a dose-dependent fashion. Rutin and harmaline effectively scavenged the hydroxyl radical and hydrogen peroxide. Treatments of oesophagitis rats with rutin and harmaline inhibited IL-1β production in the oesophagus in comparison with untreated rats, but TNF-α production was not affected by rutin and harmaline. The fMLP-induced elevation of [Ca2+]i was inhibited by rutin. 4 The results of this study suggest that rutin and harmaline may have beneficial protective effects against reflux oesophagitis by the inhibition of gastric acid secretion, oxidative stress, inflammatory cytokine production (i.e. IL-1β), and intracellular calcium mobilization in PMNs in rats.

Notes: 1 Although monumental efforts have been made to define the action sites of cough, the importance of neurotransmitter systems in the cough reflex has received limited attention. We studied the roles for four major neurotransmitters [acetylcholine, histamine, serotonin (5-hydroxytryptamine, 5-HT) and dopamine] in the modulation of the cough reflex. 2 Atropine (muscarinic cholinergic blocking agent), pyrilamine maleate (PM, histamine H1 blocker), cimetidine (histamine H2 blocker), 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT, specific 5-HT1A receptor agonist) and SCH-23390 (selective dopamine D1 receptor antagonist) were examined on the cough response to inhaled capsaicin in conscious guinea-pigs. 3 All the drugs significantly decreased the number of capsaicin-induced coughs in a dose-dependent manner. To compare the sensitivity of these drugs on cough response, we calculated the effective doses for 50% inhibition of cough (ED50) when the animals were exposed to 3 × 10−4 m capsaicin. The ED50 values were 0.03 μm kg−1 for atropine, 0.2 μm kg−1 for 8-OH-DPAT, 6.2 μm kg−1 for SCH-23390, 8.5 μm kg−1 for PM and 13.9 μm kg−1 for cimetidine. 4 These findings indicated that all these four neurotransmitters may be involved in the regulation of the cough reflex. Multiple changes of these neurotransmitters in disorders of the central nervous system might synergically affect the cough reflex.

Notes: 1 This study examined the potentiating effects of competitive antagonists of the adrenergic alpha2 receptors and of phenoxybenzamine (POB) an irreversible antagonist, on the stimulation-induced efflux of [3H]-noradrenaline in arterial tissue of rabbit. This was done to determine if the lack of concordance of efflux potentiation by antagonists with the expectations of presynaptic negative feedback theory can be attributed to increasingly successful competition from rising perineuronal transmitter concentrations, when stimulation parameters are increased, in the presence of a fixed concentration of competitive antagonist. 2 Tissues were stimulated with a fixed pulse number and frequency, to rule out confounding factors, and major alterations in the concentration of released transmitter were achieved through variations in the pulse duration. 3 Rauwolscine potentiated transmitter release less at the longest, rather than at the shortest pulse duration, and showed a potentiation of release that was indifferent to the quantities of released transmitter. This was also seen with POB although it binds covalently to the presynaptic receptor. 4 Noradrenaline inhibited stimulation-induced transmitter release confirming the presence of presynaptic alpha inhibitory sites. 5 Yohimbine potentiated transmitter release the same as did rauwolscine and POB, and protected the relevant sites against POB potentiation, confirming site identity. The combination of POB and rauwolscine had no greater effect than did either alone certifying that they acted similarly and that maximally effective concentrations of each were used. 6 Consequently, noradrenaline breakthrough of presynaptic receptor blockade does not explain the non-conforming observations made with competitive antagonists in tests of presynaptic theory.

Notes: 1 The aim of this study was to characterize the histamine receptor type mediating relaxation of the vascular bed of the nasal mucosa from the guinea-pig, and to determine the role of cyclo-oxygenase products and nitric oxide in this relaxant response to histamine. These studies were performed in isolated nasal mucosae examined in vitro to obtain potencies of histamine receptor-type selective agonists in causing vasorelaxation and to determine affinities of histamine receptor antagonists for inhibiting histamine-induced relaxation. 2 After contraction of nasal mucosae with noradrenaline, histamine caused a maximal relaxation response that was 75 ± 6% of the contraction caused by noradrenaline with a mean EC50 value of 4.3 ± 0.5 μm. Neither dimaprit (H2-receptor selective) nor R-α-methylhistamine (H3-receptor selective) caused significant relaxation of nasal mucosae. In contrast, betahistine (H1-receptor selective) caused an 81 ± 7% relaxation of noradrenaline-induced tone with an EC50 value of 15 ± 1 μm. 3 pA2 experiments were performed to obtain KB values of chlorpheniramine (H1-receptor selective) and diphenhydramine (H1-receptor selective) for blocking histamine-stimulated relaxation of nasal mucosae. KB values for chlorpheniramine (0.87 nm) and diphenhydramine (7.4 nm) were consistent with their interaction at the H1-receptor type. Additionally, neither 10 μm cimetidine (H2-receptor selective) nor 1 μm thioperamide (H3-receptor selective) had any effect on the relaxation curve for histamine. 4 In the presence of 10 μm indomethacin (cyclo-oxygenase inhibitor), histamine caused a maximal relaxation response of 73 ± 5% of the noradrenaline-induced tone with an EC50 value of 2.9 ± 0.2 μm, which was not different from control values (EC50 = 5.0 ± 0.4 μm; maximal relaxation = 71 ± 6%). In contrast, 200 μm NG-nitro-l-arginine (nitric oxide synthase inhibitor) completely inhibited histamine-induced relaxation of nasal mucosae. 5 In conclusion, data from the present study suggest only the H1-receptor type mediates relaxation of nasal mucosal blood vessels to histamine, and histamine-induced relaxation of nasal mucosae is entirely dependent on nitric oxide production.

Notes: 1 The effects of intraperitoneal (i.p.) lipopolysaccharide on vascular reactivity to noradrenaline in rat aorta under different conditions of passive tension, as well as on mortality in normotensive and hypertensive rats, were studied. 2 Concentration–response curves to noradrenaline were obtained in aorta rings, at two levels of passive tension: 3 and 0.5 g, from control and lipopolysaccharide-treated Wistar rats. Contractile responses were expressed as percentage of the maximal response to noradrenaline obtained in the beginning of the experiment at a resting tension of 2 g. The maxima were significantly larger (P 〈 0.05) at 3 g than at 0.5 g in both groups of rats: 117.8 vs. 62.3%, respectively, for control animals; 85.8 vs. 32.5%, respectively, for lipopolysaccharide-treated rats. 3 The 24-h mortality after the i.p. administration of lipopolysaccharide was lower in spontaneously hypertensive rats (1/12; 8%), when compared with control Wistar–Kyoto rats (5/11; 45%). However, mortality was higher in Wistar–Kyoto made hypertensive by 8-day administration of corticosterone (6/6; 100%). 4 We conclude that a differential sensitivity to noradrenaline of aortic smooth muscle at two different levels of passive tension is still present in lipopolysaccharide-treated animals. Chronic hypertension in SHR rats is associated with resistance to the lethal effects of lipopolysaccharide, whereas abrupt-onset hypertension induced by corticosterone leads to an increased mortality. 5 These results are compatible with the myofibrillary hypothesis, which explains vascular hyper-reactivity in chronic arterial hypertension, by postulating that a more favourable relative position (and/or proportion) for actin and myosin occurs, whereas in states of vascular hypo-reactivity, such as vasodilatory shock, the opposite phenomenon may exist.

Notes: 1 The present study aimed to evaluate the role of κ-opioid receptors at two peripheral sites, the vas deferens and the proximal colon, in κ-opioid receptor knockout mice. We investigated the role of the κ-opioid receptor in the vas deferens twitch response and in the colonic ‘off-contraction’, a rebound contractile response which follows the inhibitory response to low frequencies stimulation (10, 20, 30 Hz) and which has been suggested to ‘locally’ reproduce the contractile component of the peristaltic reflex. 2 Transmural stimulation of the vas deferens at lower frequencies (10 Hz, 10 V, 1 ms pulse trains lasting 0.5 s) evoked a contractile response that was significantly higher in the preparations from knockout mice because of lack of κ-opioid receptors than in wild type mice. A selective κ-opioid receptor agonist, U-50,488H, induced a dose-dependent inhibition of the electrically stimulated contraction in vas deferens. The percentages of reduction of the twitch response were significantly lower in knockout mice than in wild type mice after treatment with U-50,488H. The reduction of twitch response caused by U-50,488H was not reversed by administration of nor-binaltorphimine (nor-BNI) (5 × 10−6 m), a selective κ-opioid receptor antagonist, in preparations from both knockout mice and wild type mice. U-50,488H has no effect on postsynaptic adrenergic receptors, as its administration did not affect the direct contractile response to noradrenaline. 3 Transmural stimulation (5 Hz, 20 V, 2 ms pulse trains lasting 30 s) induced inhibition of spontaneous activity of colonic strips during the period of stimulation, followed by an ‘off-contraction’ after the cessation of stimulation. The statistical evaluation of the ‘off-contraction’ responses between the two strains showed no significant difference. The off-contraction, measured in specimens from knockout mice, was inhibited concentration-dependently by U-50,488H (P 〈 0.01) and significantly less than from wild type mice. 4 The effect of U-50,488H was not reversed by administration of nor-BNI (5 × 10−6 m), either in preparations from knockout mice or from wild type mice. 5 Our data may suggest that κ-opioid receptors are involved in some peripheral responses to the nerve stimulation, as indicated by the effect of U-50,488H, a selective κ-opioid receptor agonist. However, the involvement of κ-opioid receptor was also present, although less apparent, in κ -opioid receptor knockout mice, suggesting either that this drug acts not only on κ-opioid receptors but also on other receptor sites, such as κ-like receptors. An alternative interpretation can be related to a sodium channel blocking action of U-50,488H, which could explain the inhibitory effects of twitch response still present but less evident in knockout strain and the lack of effect of the antagonist nor-BNI.

Notes: 1 The effects of neuropeptide Y (NPY) upon the isolated vasculature are reviewed. 2 The vasconstrictor responses to periarterial nerve stimulation (PNS) and neurotransmission by noradrenaline (NA) and ATP are discussed and illustrated using canine isolated perfused splenic artery. 3 Modulation of the vascular responses to PNS by NPY via pre- and post-junctional NPY Y2 and Y1 receptors is discussed. 4 Evidence is presented for different α1-adrenoceptor subtypes mediating vasoconstriction to exogenous and endogenously released NA and their different locations in the neurovascular junction and extrajunctional regions. 5 Activation of NPY Y1-receptors potentiates sympathetic nerve activated α1-adrenoceptor vasoconstriction. The proposal that the postjunctional α1B adrenoceptor may be linked to the NPY Y1-receptor and is responsible for co-operation between sympathetic and NPYergic interactions in the vasculature is discussed.

Notes: 1 The parasympathetic nervous system is responsible for maintaining normal bladder function, contracting the bladder smooth muscle (detrusor) and relaxing the bladder outlet during micturition. 2 Contraction of the bladder involves direct contraction via M3 receptors and an indirect ‘re-contraction’ via M2-receptors whereby a reduction in adenylate cyclase activity reverses the relaxation induced by β-adrenoceptor stimulation. 3 Muscarinic receptors are also located on the epithelial lining of the bladder (urothelium) where they induce the release of a diffusible factor responsible for inhibiting contraction of the underlying detrusor smooth muscle. The factor remains unidentified but is not nitric oxide, a cyclooxygenase product or adenosine triphosphate. 4 Finally, muscarinic receptors are also located prejunctionally in the bladder on cholinergic and adrenergic nerve terminals, where M1-receptors facilitate transmitter release and M2 or M4-receptors inhibit transmitter release. 5 In pathological states, changes may occur in these receptor systems resulting in bladder dysfunction. Muscarinic receptor antagonists are the main therapeutic agents available for treatment of the overactive bladder, but whether their therapeutic effect involves actions at all three locations (detrusor, prejunctional, urothelial) has yet to be established.

Notes: 1 The aim of this study was to examine whether sodium nitroprusside (SNP)-induced relaxation of rat fundus longitudinal smooth muscle involves ryanodine-sensitive Ca2+ release. 2 SNP (300 nm–30 μm) elicited concentration-dependent relaxation of precontracted (1 μm carbachol) rat fundus, an effect almost abolished by the selective guanylyl cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ, 10 μm). 3 SNP-mediated relaxations were almost abolished by 10 μm ryanodine. 4 SNP-mediated relaxations were also reduced by either 1 μm apamin (a selective small conductance Ca2+-sensitive K+ channel, SKCa, inhibitor) or the selective L-type Ca2+ channel inhibitor, nicardipine (3 μm). 5 SNP-induced relaxations were insensitive to 1 mm tetraethylammonium chloride (an inhibitor of large-conductance Ca2+-sensitive K+ channels) and 1 μm glibenclamide (an ATP-sensitive K+ channel inhibitor). 6 These data suggest that SNP-mediated fundus relaxation occurs via a cGMP-mediated and ryanodine-sensitive mechanism which requires, at least in part, SKCa and L-type Ca2+ channel activity.

Notes: 1 The aim of this work was to investigate the effect of acute and chronic exposure of rat pheochromocytoma (PC12) cells to elevated cAMP, using forskolin, dibutyryl-cAMP (db-cAMP) or isobutylmethylxanthine (IBMX), on endogenous dopamine content and release and on [3H]-1-methyl-4-phenylpyridinium ([3H]-MPP+) uptake and release, under basal conditions and under KCl-stimulation. 2 Cultured PC12 cells synthetized and accumulated large amounts of dopamine, but not noradrenaline or adrenaline. The release of dopamine by the cells was markedly increased in response to 50 mm KCl. 3 Acute and chronic treatment of the cells with forskolin (30 μm), but not IBMX (100 μm), slightly increased the spontaneous release of dopamine and significantly decreased the release induced by 50 mm KCl. 4 Chronic treatment of the cells with forskolin (30 μm), but not IBMX (100 μm), markedly decreased the cellular content of dopamine. 5 Cultured PC12 cells removed and accumulated [3H]-MPP+, which, similarly to dopamine, was released by KCl. 6 Acute treatment of the cells with forskolin (30 μm) or db-cAMP (2.5 mm), but not IBMX (100 μm), slightly increased the spontaneous release, but did not affect KCl-induced release of [3H]-MPP+. On the other hand, chronic treatment of the cells with forskolin produced, on [3H]-MPP+, similar effects to those obtained for dopamine. 7 Acute and chronic treatment of the cells with reserpine (50 nm) produced similar results to those obtained with forskolin on either dopamine or [3H]-MPP+ handling. 8 In conclusion, cAMP, similarly to reserpine, increases the spontaneous release and decreases the KCl-induced release of [3H]-MPP+ and dopamine. This suggests that cAMP impairs the vesicular monoamine transporter.

Notes: 1 The aim of this study was to characterize the adenosine receptor mediating vasodilation in the microvasculature of the hamster cheek pouch in vivo. A range of adenosine agonists was used including N6-cyclopentyladenosine (CPA) (A1 agonist), 5′-N-ethylcarboxamidoadenosine (NECA) (non-selective), 2-chloroadenosine (2CADO) (non-selective), 2-p-(2-carboxyethyl)-phenethylamino-5′-N-ethylcarboxamidoadenosine (CGS 21680) (A2A agonist), N6-(3-iodobenzyl)-adenosine-5′-N-methyluronamide (IBMECA) (A3 agonist) and adenosine, as well as the adenosine antagonists 8-sulphophenyltheophylline (8-SPT) (A1/A2 antagonist), 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) (A1 antagonist) and 4-(2-[7-amino-2-(2-furyl)[1,2,4]-triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM 241385) (A2A antagonist). 2 All the adenosine analogues used induced vasodilation at concentrations between 10 nm and 1 μm, and the potency order was NECA 〉 CGS 21680 〉 2CADO 〉 CPA=IBMECA 〉〉 adenosine, indicating an action at A2A receptors. 8-SPT (50 μm) antagonized vasodilator responses to NECA with an apparent pKB of 5.4, consistent with an action at A1 or A2 receptors and confirming that A3 receptors are not involved in this response. 3 DPCPX (10 nm) had no effect on vasodilation evoked by NECA, suggesting that this response was not mediated via A1 receptors, while ZM 241385 (10 nm) antagonized dilator responses to NECA with an apparent pKB of 8.9 consistent with an action via A2A receptors. 4 Overall these results suggest that adenosine A2A receptors mediate vasodilation in the hamster cheek pouch in vivo.

Notes: 1 The hypothesis that the non-adrenergic, non-cholinergic excitatory (NANC-e) innervation is involved in the induction of asthma and that antagonists of NANC-e neurotransmitter could reduce bronchoconstriction during asthma was tested. 2 The first objective was to identify the neurotransmitter(s) of NANC-e innervation from a group of selected putative neurotransmitters. The second objective was to use the antagonist of the identified neurotransmitter(s) to determine its effectiveness against bronchoconstriction to ovalbumin (OVA) in sensitized guinea-pigs. 3 Neurotransmitter identification was performed using the ‘tracheal pouch’, a surgical preparation established for demonstrating NANC innervation, in anaesthetized guinea-pig airways. A segment of trachea was cannulated and clamped at one end and the other end was connected to a pressure transducer. The stump of the trachea was connected to a ventilator to keep the blood gas values within the normal range. The vagus nerve and the sympathetic nerves were isolated bilaterally and cut. The left carotid artery was cannulated to monitor blood pressure and for sampling blood for blood gas analysis. The jugular vein was cannulated for administration of test agents. 4 Both NANC-e and NANC-i (inhibitory) control responses of airways were obtained by bilateral vagal stimulation after complete autonomic blockade with atropine, propranolol and prazosin. The relaxation of the tracheal pouch was indicative of the NANC-i response and the increase in insufflation pressure of the ventilated peripheral airways was due to NANC-e stimulation. 5 The involvement of the putative neurotransmitters such as neurokinin-A (NK-A), histamine, serotonin and endothelin (ET) was investigated by using the respective antagonists, MEN-10376, pyrilamine maleate, cyproheptadine hydrochloride, and two ET receptor antagonists (BQ-123 and IRL-1038), respectively. The antagonists were administered at the dose rate of 4 mg kg−1 i.v. which was determined from preliminary studies by testing against the respective agonists. 6 MEN-10376 (neurokinin-2 receptor antagonist) significantly inhibited the insufflation pressure (peripheral airway pressure) increase caused by NANC-e stimulation. MEN-10376 also inhibited the fall in blood pressure caused by bilateral vagal stimulation. The 5-HT antagonist, cyproheptadine, significantly enhanced the NANC-e response. 7 After identifying the NANC-e neurotransmitter as NK-A, the effectiveness of its antagonist, MEN-10376, was evaluated for its ability to attenuate the increase in insufflation pressure (bronchoconstriction) induced in guinea-pigs sensitized by OVA. Guinea-pigs were sensitized to OVA (200 mg i.p.) and 10 days later prepared for the determination of tracheal pouch and insufflation responses to 100 μg of OVA administered i.v. (challenge dose). This caused an increase in insufflation pressure in the presence of adrenergic and cholinergic blockade, which was significantly attenuated by MEN-10376. 8 These studies indicated that neurokinin-2 receptors were involved in the vagally mediated efferent neurotransmission of NANC-e and that NANC-e plays a role in allergen-induced bronchoconstriction.

Notes: 1 The effect of WAY100635 (N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl) cyclohexanecarboxamide trihydrochloride), a 5-HT1A receptor antagonist, on cardiovascular function was studied. 2 The i.v. injection of WAY100635 dose-dependently decreased blood pressure in anaesthetized rats; while in pithed rats WAY100635 (1 mg  kg−1) displaced the phenylephrine pressor effect. 3 WAY100635 antagonized phenylephrine-induced contraction in rabbit and rat aorta (pA2 of 6.88 and 7.93 and Schild slopes of −0.83 and −1.21, respectively); while in rat caudal artery pKB was 7.45 and the Schild slope of −0.56, suggesting a complex interaction in this vessel. 4 The results show that WAY100635 induced hypotension in the anaesthetized rat and suggest that this effect could be partially explained by antagonism of vascular α1-adrenoceptors.

Notes: 1 Purinergic transmission from sympathetic nerves in the guinea-pig vas deferens was monitored using intracellular recording techniques. Stimulation of the hypogastric nerve with trains of 15 pulses at 1 Hz evoked excitatory junction potentials (EJPs) which increased in amplitude from the first pulse and reached a maximum after 6–8 pulses. 2 Caffeine (3 and 10 mm), depolarized cells by 5–10 mV and increased the amplitude of the first few EJPs in each train but reduced the maximum amplitude of EJPs late in the train. 3 The adenosine receptor antagonist 8-p-sulphophenyl-theophylline (8-SPT; 30 μm) had no effect on either the resting membrane potential or the EJP amplitude; however, at 100 μm it reduced the amplitude of all EJPs by 5–10%. 4 Adenosine (10 and 30 μm) reduced the amplitude of EJPs in a concentration-dependent manner. The inhibitory effect of adenosine on EJP amplitude was prevented by pretreatment with either caffeine (3 mm) or 8-SPT (30 μm). 5 Ryanodine (30 μm) did not alter EJP amplitude and did not inhibit the enhancement of the first EJP by caffeine (3 mm). Incubation of the tissue with the cell permeable calcium chelator 1-2-bis(o-aminophenoxy)ethane-N,N-N′,N′-tetraacetic acid (BAPT-AM) resulted in a depression of EJP amplitude and a longer time to reach maximum amplitude. In cells that had been exposed to BAPT-AM, caffeine 3 mm still increased amplitude of EJP early in the train. 6 The phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX; 500 μm), hyperpolarized cells and increased the amplitude of EJP throughout the train of stimulation. In the presence of IBMX, caffeine 3 mm still depolarized the cells and enhanced the EJP early in the train of stimulation. 7 The findings in this study confirm that caffeine and 8-SPT are effective inhibitors of the actions of adenosine. However, caffeine has an additional action to enhance EJP early during a train of stimulation, which cannot be attributed to blockade of adenosine receptors, but which may be related to inhibition of phosphodiesterase.

Notes: 1 The β3-adrenoceptor agonist ZD 7114, like the non-selective β-adrenoceptor agonist isoprenaline, but unlike the β1-adrenoceptor agonist dobutamine and the β2-adrenoceptor agonist salbutamol, produced an increment on mouse embryonic fibroblast proliferation. The half-maximal stimulation of cell growth occurred at substantially lower concentrations with the β3-adrenoceptor agonist (EC50: 5.5 × 10−8 m) than with isoprenaline (EC50: 1.25 × 10−6 m). 2 The selective β3-adrenoceptor antagonist SR 5923 OA prevented the β3-stimulated fibroblast proliferation. Conversely, practolol and butoxamine did not prevent fibroblast growth. 3 Additionally, a decrease of cAMP was obtained in fibroblasts cells upon stimulation with isoprenaline and ZD 7114. 4 The expression of β-adrenoreceptors on fibroblast cells was also studied by radioligand binding. The Ki values in the presence of β1- and β2-adrenoceptor antagonist was two-fold higher than the Ki values for β3 adrenoceptor antagonist indicating the presence of A3-receptor subtype. 5 Inhibitors of different intracellular coupling pathways including phospholipase C (U 73122), protein kinase C (staurosporine), calcium/calmodulin (trifluoroperazine) and calcium channel (verapamil), prevented the stimulatory actions of the selective β3-adrenoceptor agonist ZD 7114. 6 The presence of β3-adrenoceptors on embryonic mouse fibroblast cells may play a role in the modulation of cell growth and biologic activity. The mechanism by which ZD 7114 triggers cell proliferation and function, involves the activation of phospholipase C, PKC, calcium/calmodulin and the influx of calcium.

Notes: 1 Cytokines may parallel or regulate the beneficial effects of β-adrenoceptor antagonist treatment observed in chronic heart failure (CHF) patients. Therefore, this study was performed in order to investigate alterations of cytokine levels in β-blocker-treated patients suffering from CHF. 2 We investigated plasma cytokine levels in eight healthy controls and 12 CHF patients. The patients were treated with standard medication (CHFstd) or with standard medication and additional β1-blocker metoprolol (CHFmet). Interleukin-(IL)-1α, IL-1β, IL-1 receptor antagonist (IL-1ra), IL-2, IL-6, IL-8, IL-10, tumor necrosis factor-α (TNF), soluble TNF receptor type 1 (sTNF-R1), sTNF-R2, and sCD14 were measured by ELISA. 3 IL-1α and IL-1β were not detectable in any of the tested groups. IL-2, TNF, or sCD14 were not altered as compared with healthy control subjects. CHFstd patients expressed enhanced IL-1ra, IL-6, IL-8, IL-10, sTNF-R1 and sTNF-R2. In CHFmet patients IL-1ra, IL-6 and IL-8 remained at the same level. In contrast, sTNF-R1 levels were significantly reduced, although not to control, whereas the sTNF-R2 and IL-10 were reduced to control levels. 4 The cAMP levels of mononuclear cells – recalculated for the patients included in this study from previous work [Werner et al. (2001). Basic Res. Cardiol., 96, 290]– correlated inversely with the sTNF-R2 data (Pearson, r=−0.46; P=0.041; Spearman, r=−0.64, P=0.002). 5 The present data indicate an interaction of the neurohumoral and the cytokine system in CHF patients at the cAMP level. Thus, measurement and correlation of sTNF-R2 and cAMP may provide a tool useful during investigation of β-blocker therapy.

Notes: 1 There is good evidence that β-blockers improve ventricular function, disease progression and survival in patients with left ventricular systolic dysfunction. The aim of this study was to determine the effects of atenolol therapy on the sympathetic nervous system at rest and after ergometric exercise, on left ventricular function and on baseline plasma atrial natriuretic factor (ANF) in ambulatory patients with chronic heart failure (CHF). 2 Twenty-two patients [left ventricular ejection fraction (LVEF) 〈36%; New York Heart Association II–III] were studied before atenolol treatment. Because of cardiac events (new Hospital admission or death) only 13 patients completed 1 year of treatment. Baseline noradrenaline (NE) concentrations were similar in patients and controls while ANF was higher in patients than in controls (328 ± 35 pg ml−1vs. 37 ± 3 pg ml−1; P 〈 0.01). 3 Patients with events showed higher NE (540 ± 87 pg ml−1vs. 303 ± 44 pg ml−1; P 〈 0.01) and ANF (460 ± 70 pg ml−1vs. 291 ± 44 pg ml−1; P 〈 0.03) at rest; and greater NE response to exercise (2.003 ± 525 pg ml−1vs. 694 ± 121 pg ml−1; P 〈 0.005). Atenolol treatment improved LVEF (19.5 ± 1.9%vs. 33 ± 3.9%; P 〈 0.001), increased exercise tolerance (9 ± 3.2 min vs. 17 ± 4.8 min; P 〈 0.001) and decreased plasma ANF (292 ± 42 pg ml−1vs. 133 ± 35 pg ml−1; P 〈 0.01). 4 Reduced basal dihydroxyphenylglycol (DHPG)/NE ratio (3.4 ± 0.46 vs. 4.3 ± 0.35; P 〈 0.01) was observed in patients compared with healthy volunteers. Atenolol increased DHPG plasma levels (1.398 ± 129 pg ml−1vs. 913 ± 86 pg ml−1; P 〈 0.005) but the DHPG/NE ratio during exercise was not modified after treatment, suggesting that re-uptake of released NE is not changed by β-blocker treatment. 5 In conclusion, the fact that atenolol treatment improves ventricular dysfunction and clinical status without changing plasma NE levels in CHF patients, suggests that plasma NE is a poor surrogate measurement for cardiac sympathetic activity in this pathology. In addition, decrease in plasma ANF produced by atenolol treatment may reflect the improvement of ventricular function.

Notes: 1 The aim of the study was to investigate the role of the α1D-adrenoceptor in α1-adrenoceptor-induced contraction of human prostate by means of protection experiments. 2 Responses of human prostate strips to noradrenaline were recorded, along with responses of rat aorta and vas deferens, tissues possessing predominantly α1D- and α1A-adrenoceptors respectively, for comparison. α1-adrenoceptors were then inactivated by incubation with the irreversible antagonist phenoxybenzamine. In some tissues α1A- or α1D-adrenoceptors were ‘protected’ from inactivation by incubation in the presence of the selective α1A- or 1D-adrenoceptor antagonists 5-methylurapidil and BMY 7378 before recording further responses to noradrenaline. 3 Phenoxybenzamine reduced the maximum noradrenaline-induced response and the potency of noradrenaline in all tissues. In rat vas deferens and human prostate, 5-methylurapidil protected α1A-adrenoceptors in a concentration–dependent manner. In rat aorta, 10 nm BMY 7378 almost fully protected α1D-adrenoceptors. However, concentrations of BMY 7378 up to 30-fold higher failed to protect receptors in the human prostate. 4 These results suggest that in human prostate functional α1D-adrenoceptors do not contribute to noradrenaline-induced contractile responses.

Notes: 1 The aim of this study was to determine if stimulation of duodenal motility by duodenal fluid distension or by administration of carbachol, activates the sphincter of Oddi-duodenal reflex, in an in vitro preparation from the Australian possum. 2 Duodenal distension was achieved by infusion of Krebs solution (0–8 cm H2O). In separate experiments, the sphincter of Oddi (SO) was partitioned from the duodenum and carbachol (10−7−5 × 10−6 m) added to the duodenal compartment. 3 Fluid distension increased duodenal motility to 120–600% of control activity. These treatments induced increased SO motility (to 120–390% of control) in six preparations, reduced activity (to 60% of control) in one and no response in another. 4 Addition of carbachol to the duodenal compartment resulted in increased duodenal motility. SO motility was increased in seven preparations, reduced in another two and no response were evoked in two others. All SO responses were blocked by tetrodotoxin pretreatment. 5 These data suggest that the SO receives inputs from duodenal mechano and/or stretch receptors resulting in excitatory or inhibitory responses, with the excitatory response dominating. These findings support the role for the SO-duodenal reflex in preventing duodeno-biliary/pancreatic reflux during periods of elevated duodenal activity.

Notes: 1 A triple organ bath method allowing the synchronous recording of the motor activity of the circular muscle layer belonging to the oral and anal segments of guinea-pig small intestine adjacent to an electrically stimulated middle segment was developed to study the ascending and descending reflex motor responses. 2 Electrical field stimulation (0.8 ms, 40 V, 5 Hz, 10 s) applied to the middle part of the segments elicited tetrodotoxin (1 μm)-sensitive ascending and descending contractile responses of the nonstimulated parts, oral and anal, respectively. The ascending contraction was more pronounced as compared with the descending contraction. 3 In the presence of phentolamine (5 μm), propranolol (5 μm) and atropine (3 μm) a significant decrease in the amplitude of the ascending contraction was seen and a descending relaxation, instead of a contraction was observed. 4 Met-enkephalin applied at a single concentration (0.1 μm) or cumulatively (0.001–1 μm) inhibited both non-adrenergic non-cholinergic (NANC) descending relaxation and ascending contraction with similar efficacy but different potency, IC50 being 5.9 ± 0.3 and 39.0 ± 4 nm, respectively. Naloxone (0.5 μm) prevented the effects of Met-enkephalin. 5 L-NNA (0.5 mm), an inhibitor of nitric oxide synthesis, increased the ascending contraction and strongly reduced but not abolished the descending relaxation. l-Arginine (0.5 mm) restored the motor responses to the initial level in l-NNA-pretreated preparations, d-Arginine (0.5 nm) had no effects. 6 Met-enkephalin (0.1 μm) depressed the l-NNA-dependent increase of the ascending contraction and failed to change the l-NNA-resistant part of the descending relaxation. 7 Met-enkephalin did not alter spontaneous NANC mechanical activity. SNP (1 or 10 μm), an exogenous donor of nitric oxide, caused a concentration-dependent relaxation. The effects of SNP persisted in Met-enkephalin (0.1 μm)-pretreated preparations. 8 NANC reflex ascending contraction and descending relaxation were synchronously induced by a local nerve stimulation indicating a functional coactivation of NANC orally projected excitatory and anally directed inhibitory pathways. Acting prejunctionally, Met-enkephalin provided a negative controlling mechanism inhibiting both ascending and descending, mainly nitric oxide mediated, reflex responses. A higher sensitivity of the descending relaxation to Met-enkephalin was observed suggesting an essential role of opioid(s) in reducing the efficacy of descending motor activity.

Notes: 1 The present study was designed to investigate the secretion of catecholamines (CA) evoked by stimulation of cholinergic receptors and membrane depolarization from the isolated perfused adrenal gland of spontaneously hypertensive rats (SHR) and normotensive Wistar–Kyoto rats (WKYR) at adult age. 2 The wet weight of adrenal gland in SHR was greater than that in WKYR. The CA releasing responses evoked by acetylcholine (5.32 × 10−3 m), and high potassium (5.6 × 10−2 m), a membrane depolarizer, were significantly lower in WKYR than in SHR. 3 The secretory responses of CA evoked by DMPP (10−4 m for 2 min), a selective agonist of neuronal nicotinic receptors, and McN-A-343 (10−4 m for 2 min), a selective agonist of neuronal muscarinic receptors, were also significantly lower in WKYR than in SHR. 4 The CA release evoked by Bay-K-8644 (10−5 m), a dihydropyridine-sensitive Ca2+ channel activator, and cyclopiazonic acid (10−5 m), a selective inhibitor of Ca2+-ATPase in the endoplasmic reticulum, were also significantly greater in SHR than WKYR. 5 Taken together, these experimental results demonstrate that the CA secretion evoked by stimulation of cholinergic (nicotinic and muscarinic) receptors as well as membrane depolarization is enhanced more greatly in the perfused adrenal glands of SHR than in those of WKYR. It is suggested that the augmented CA release in SHR compared with WKYR was involved in essential hypertensive pathogenesis.

Notes: 1 The contractile capacity of smooth muscle cells depends on the cytoskeletal framework of the cell. The aim of this study was to determine the functional importance of both the actin and the tubulin components of the cytoskeleton in contractile responses of the bovine isolated iris sphincter muscle. 2 In each preparation, two contractions to the muscarinic agonist carbachol were obtained. The maximum responses of the first contractions were taken as 100%. The second contractions to carbachol were elicited in the presence of either cytochalasin B (50 and 5 μm), an inhibitor of the actin cytoskeleton, or colchicine (100 μm), an inhibitor of the tubulin cytoskeleton (30 min incubation). 3 Cytochalasin B, at a concentration of 50 μm, significantly decreased the contractions induced by carbachol, with the maximum response reduced to 21.8 ± 6.6% (n = 12) of the initial maximum. The maximal contractions to carbachol in the presence of colchicine reached 96.2 ± 7.9% (n = 9) of the initial contraction, which was not significantly different from control second responses to carbachol with neither drug present, which reached 113.3 ± 7.6% (n = 7). 4 The effect of cytochalasin B was dose-dependent, since at a lower concentration of 5 μm, the drug decreased the maximum contraction to carbachol to 60.3 ± 8.8% (n = 6). The effect of cytochalasin B was at least partially reversible, since after the use of the higher concentration of 50 μm, contractions to carbachol increased to 62.3 ± 15.5% (n = 4) of the maximal response, after 1 h repeated washing of the preparations. 5 Cytochalasin D, at a concentration of 50 μm, completely abolished the contractions induced by carbachol (n = 4). 6 These findings suggest that in bovine iris sphincter muscle, contractions to carbachol are highly dependent, from a functional point of view, on actin polymerization, and not, to any important degree, on the polymerization of tubulin.

Notes: Summary 1 GABAB1 receptor subunit knockout mice were generated and the effects of the GABAB receptor agonist, baclofen, were evaluated within the peripheral nervous system (PNS) of wildtype (+/+), heterozygote (+/–) and knockout (–/–) animals. For this purpose, neuronally-mediated responses were evoked in both the isolated ileum and urinary bladder, using selective electrical field stimulation (EFS). 2 In ileum resected from 4–8-week-old-mice, low frequencies of EFS (0.5 Hz) evoked irregular muscle contractions which were prevented by atropine 1 μM and reduced by baclofen (33.4 ± 5.6%, 100 μm). The latter effect was antagonized by the GABAB receptor antagonist CGP54626 0.2 μm. Baclofen 100 μm did not affect contractions of similar amplitude induced by carbachol, indicating that the ability of baclofen to inhibit cholinergic function in mouse ileum may be due to an action at prejunctional GABAB receptors. 3 To avoid the development of grand mal seizure by GABAB1 (–/–) mice, a behaviour observed when the mice were greater than 3 weeks old, it was necessary to study the effects of this knockout in 1–3-week-old-animals. However, at this age, EFS at 0.5 Hz did not evoke robust muscle contractions. Consequently we used EFS at 5 Hz, which did evoke cholinergically mediated contractions, found to be of similar amplitude in (+/+) and (+/–) mice, of both 1–3 weeks and 4–8 weeks of age. At this frequency of EFS, baclofen reduced the amplitude of the evoked contractions [n=6 (+/+) and n=5 (+/–), IC50 19.2 ± 4.8 μm) and this effect was greatly reduced in the presence of CGP54626 0.2 μm. 4 In urinary bladder from 1–3-week-old-mice, using higher frequencies of EFS to evoke clear, nerve-mediated contractions (10 Hz), baclofen 10–300 μm concentration-dependently inhibited contractions in (+/+) mice (IC50 9.6 ± 3.8 μm). This effect was inhibited by CGP54626 (0.2 μm, 46.2 ± 13.6% inhibition, 300 μm baclofen n=7) a concentration which, by itself, had no effect on the EFS-evoked contractions. 5 The effects of baclofen in both ileum and urinary bladder were absent in the GABAB1 receptor subunit (–/–) mice; however, responses to EFS were unaffected in (–/–) when compared to the (+/+) mice. 6 Our data suggest that, as in the central nervous system (CNS), the GABAB1 receptor subunit is an essential requirement for GABAB receptor function in the enteric and PNS. As such, these data do not provide a structural explanation for the existence of putative subtypes of GABAB receptor, suggested by studies such as those in which different rank-orders of GABAB agonist affinity have been reported in different tissues.

Notes: 1 Type 2 diabetes is associated with diverse oral pathologies in which salivary flow reduction is one of the causes of these oral abnormalities. Scarce literature exists regarding noradrenergic transmission and adrenergic-induced salivary flow in submaxillary and parotid glands of type 2 diabetic rats. 2 We studied noradrenergic transmission as well as the secretory response to α1- and β-adrenoceptor stimulation in the parotid and submaxillary glands of type 2 diabetic rats. 3 Diabetic rats exhibited diminished neuronal uptake, release and endogenous content of noradrenaline (NE) in both salivary glands. Further, NE synthesis was also diminished accompanied by decreased tyrosine hydroxylase activity. Salivary flow responses to α1-(methoxamine) and β-(isoprenaline) adrenoceptor stimulation were reduced in the submaxillary as well as the parotid glands of diabetic rats. 4 Our results suggest that the reduction of noradrenergic transmission in the salivary glands of type 2 diabetic rats is in part responsible for the diminished salivary flow evoked by α1- and β-adrenergic stimulation. Reduced noradrenergic activity may contribute to the pathophysiology of oral abnormalities in diabetic patients.

Notes: 1 We investigated the effects of exogenously applied steroids and endogenously released cortisol on catecholamine (CA) secretion induced by cholinergic agonists in perfused guinea-pig adrenal glands. 2 Acetylcholine (ACh) and electrical stimulation induced CA secretion, which was reversibly inhibited by cortisol. Adrenocorticotropic hormone (ACTH) increased the concentration of cortisol in the perfusion effluent and partly inhibited the secretory response to ACh. 3 Cortisol or aldosterone dose-dependently inhibited secretory responses to nicotine and muscarine. These inhibitory effects were not antagonized by mifepristone and spironolactone, respective cortisol and aldosterone receptor blockers. 4 Dexamethasone, cortisone, corticosterone, 11-deoxycortisol, 11-deoxycorticosterone, prednisolone and cholesterol inhibited nicotine-evoked CA secretion. The secretory response to muscarine was inhibited by these compounds except for dexamethasone and prednisolone. 5 Dexamethasone, cortisol and aldosterone had no effect on CA secretion induced by high KCl. 6 These results suggest that steroids affect nicotinic and muscarinic ACh receptor-mediated responses through distinct mechanisms, and that cortisol released from the adrenal cortex inhibits CA secretion from the adrenal medulla.

Notes: Summary 1 Bradykinin (BK) is a nine amino acid peptide (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg) formed from the plasma precursor kininogen during inflammation and tissue injury. The actions of BK are mediated by G protein-coupled cell surface receptors, designated B1 and B2. 2 BK has a plethora of effects in the airways including bronchoconstriction, bronchodilation, stimulation of cholinergic and sensory nerves, mucus secretion, cough and oedema resulting from promotion of microvascular leakage. These airway effects are mediated in the main by the B2 receptor subtype. 3 BK acts mainly indirectly, primarily through airway nerve activation, but also by the release of prostanoids, thromboxanes and nitric oxide (NO). 4 Airway responses to BK have been studied in detail in guinea-pigs, mice, sheep and rats. This review describes the effects of BK in these species and draws comparison with its effects in normal humans and patients with respiratory diseases. 5 Despite its many and varied effects in the airways of animals and man, the exact contribution of BK to airways disease remains unclear.

Notes: 1 The purpose of this study was to demonstrate that the adrenergic nervous system regulates the in vivo choline levels in the mouse major salivary glands. 2 Methoxamine (α1-adrenoceptor agonist, 2.5–20 mg kg−1, s.c.) elevated choline levels dose-dependently and the effect of methoxamine (10 mg kg−1) was completely inhibited by the α-adrenoceptor antagonist phentolamine (5 mg kg−1, i.p.) but not by the β-adrenoceptor antagonist propranolol (3 mg kg−1, i.p.). 3 In contrast, isoprenaline (β-adrenoceptor agonist 0.25–20 mg kg−1, s.c.) lowered choline levels and the effect of isoprenaline (2 mg kg−1) was inhibited by propranolol, but not by phentolamine. 4 Noradrenaline (1–4 mg kg−1, s.c.) manifested both the α- and β-adrenergic actions depending on its dose. Noradrenaline at 1–2 mg kg−1, lowered choline levels and the effect of noradrenaline (1 mg kg−1) was inhibited by propranolol, but not by phentolamine. On the other hand, noradrenaline (4 mg kg−1) elevated choline levels and the effect was blocked by phentolamine, but not by propranolol. 5 Tyramine (5–80 mg kg−1, s.c.) elicited the release of noradrenaline from sympathetic nerve terminals and induced essentially the same effects on the choline levels as noradrenaline. Tyramine (10 mg kg−1) lowered choline levels and the effect was inhibited by propranolol, but not by phentolamine. However, tyramine (80 mg kg−1) elevated choline levels and the effect was inhibited by phentolamine, but not by propranolol. 6 These results suggest that choline levels in the salivary glands may be under separate α- and β-adrenergic control and suggest a possibility that the neurotransmitter noradrenaline released for sympathetic nerve terminals can manage the dual control of choline levels in some autonomic organs in a characteristic dose-dependent manner.

Notes: 1 The purpose of this study was to investigate the protective effects of defibrotide, a single-stranded polydeoxyribonucleotide, on ischaemia–reperfusion injury to the liver using a rat model. 2 Ischaemia of the left and median lobes was created by total inflow occlusion for 30 min followed by 60 min of reperfusion. Hepatic injury was assessed by the release of liver enzymes (alanine transferase, ALT and lactic dehydrogenase, LDH). Hepatic oxidant stress was measured by superoxide production, lipid peroxidation and nitrite/nitrate formation. Leukocyte–endothelium interaction and Kupffer cell mobilization were quantified by measuring hepatic myeloperoxidase (MPO), polymorphonuclear leukocyte adherence to superior mesenteric artery (SMA) and immunostaining of Kupffer cell. 3 Defibrotide treatment resulted in a significant inhibition of postreperfusion superoxide generation, lipid peroxidation, serum ALT activity, serum LDH activity, MPO activity, serum nitrite/nitrate level, leukocyte adherence to SMA, and Kupffer cell mobilization, indicating a significant attenuation of hepatic dysfunction. 4 A significant correlation existed between liver ischaemia/reperfusion and hepatic injury, suggesting that liver ischaemia/reperfusion injury is mediated predominantly by generation of oxygen free radicals and mobilization of Kupffer cells. 5 We conclude that defibrotide significantly protects the liver against liver ischaemia/reperfusion injury by interfering with Kupffer cell mobilization and formation of oxygen free radicals. This study provides strong evidence that defibrotide has important beneficial effects on acute inflammatory tissue injury such as that occurring in the reperfusion of the ischaemic liver.

Notes: 1 The effects of BIIE 0246, a novel and non-peptide neuropeptide Y (NPY) Y2 receptor antagonist on sympathetic vasoconstriction of the canine splenic artery were investigated. 2 The vasoconstrictor response to periarterial electrical nerve stimulation was described to be a double peaked vasoconstriction consisting of an initial transient, dominantly P2X purinoceptor-mediated constriction followed by a prolonged, mainly α1 adrenoceptor-induced response. 3 BIIE 0246 at a concentration of 0.1–1 μm dose-dependently potentiated double peaked constrictions at low frequencies (1 and 4 Hz), whereas at high frequency (10 Hz), it failed to affect these responses. BIIE 0246 (1 μm) also enhanced double peaked responses even in the presence of rauwolscine (0.1 μm). NPY (13–36) (1–100 nm), a selective Y2 receptor agonist reduced these two peaked responses in a dose-related manner. The vasoconstriction to noradrenaline (0.1–10 nmol) or adenosine triphosphate (0.01–1 μmol) was not significantly influenced by either 1 μm BIIE 0246 or 100 nm NPY (13–36). Exposure of tissues to 1 μm BIIE 0246 almost completely prevented the suppression of double peaked constrictions by NPY (13–36) (10 nm) or by NPY (10 nm). 4 We conclude that NPY inhibits sympathetic purinergic and adrenergic vasoconstrictions through an activation of prejunctional Y2 receptor subtype in the neurovascular junction of the canine splenic artery.

Notes: Summary 1 The agonistic and antagonistic effects of labetalol, the α1- and β-adrenoceptor antagonist, were studied on β3-adrenoceptors in the guinea-pig gastric fundus. 2 Labetalol caused a concentration-dependent relaxation with a pD2 value of 5.58 ± 0.09 and an intrinsic activity of 0.64 ± 0.06, which was not affected by pretreatment with both the selective β1-adrenoceptor antagonist, (±)-atenolol (100 μM), and the selective β2-adrenoceptor antagonist, (±)-butoxamine (100 μM). 3 However, the non-selective β1-, β2- and β3-adrenoceptor antagonist, (±)-bupranolol (3–30 μM), shifted the concentration–response curve of labetalol to the right (pA2 value=5.97 ± 0.08). 4 In the presence of (±)-atenolol (100 μM) and (±)-butoxamine (100 μM), relaxations to catecholamines [(−)-isoprenaline, (−)-noradrenaline and (−)-adrenaline], to the selective β3-adrenoceptor agonist, BRL37344, and to the non-conventional partial β3-adrenoceptor agonist, (±)-CGP12177A, were weakly antagonized by labetalol (10 μM). 5 These results indicate that labetalol, the arylethanolamine, acts as a partial agonist on β3-adrenoceptors in the guinea-pig gastric fundus.

Notes: Fishing hooks retained from previous capture events were found in 6 of 211 blue sharks, Prionace glauca (L.), landed in the summers of 1999 and 2000 by recreational fishermen off Long Island (New York, USA). The hooks were embedded within the distal oesophagus (n=3), or perforated the gastric wall (n=3) and lacerated the liver (n=2). The hooks were surrounded by excessive fibronecrotic tissue which ablated the normal anatomical structures and in the three sharks with oesophageal hooks caused partial luminal obstruction. Accompanying lesions included oesophagitis, gastritis, hepatitis and proliferative peritonitis. Aeromonas sp. and Vibrio sp. were isolated from the peritoneal fluid of one shark with peritonitis and intralesional bacteria were seen on histological examination in all sharks. This is the first report of the prevalence and pathology of retained fishing hooks in a large number of wild-caught sharks.

Notes: White-clawed crayfish, Austropotamobius pallipes, were endemic to the Nant watershed, Ardéche, France, until they were extirpated by epizootic mortality at the beginning of the twentieth century. A. pallipes were successfully reintroduced to the Nant watershed in the middle of the twentieth century. However, epizootic mortality was observed in the Nant watershed in the summer of 2000 during which time A. pallipes was extirpated from downstream regions. Dead and moribund crayfish were again detected in several episodes in summer 2001 and by October the range of A. pallipes was reduced to the headwaters of just one of the three streams in the watershed. Water quality for the watershed in summer 2001 was appropriate for crayfish habitation. Bacteriology and mycology on A. pallipes collected during several of the mortality episodes in 2001 failed to reveal a cause. However, histopathology revealed a high occurrence of intranuclear eosinophilic inclusions in hepatopancreatocytes of A. pallipes. The nuclei were hypertrophic and contained bacilliform virions consisting of a cylindrical nucleocapsid surrounded by a trilaminar envelope. Virions in section were approximately 63 × 258 nm and nucleocapsids were approximately 52 × 225 nm. It is unclear whether the intranuclear bacilliform virus was the cause of the mortality episodes or was a contributor to a disease complex involving one or several other undetected pathogens.

Notes: Freshwater bryozoans have recently been identified as hosts of Tetracapsula bryosalmonae, the causative agent of proliferative kidney disease (PKD). To gain insight as to which bryozoans are consistently important hosts for T. bryosalmonae, we present the results of a broad scale survey of bryozoans in European and North American sites upstream from PKD outbreaks in feral, stocked and farmed salmonids. The bryozoan genera most commonly found in association with disease outbreaks are Fredericella and Plumatella, and the most common species are F. sultana (Blumenbach), F. indica Annandale, and P. emarginata Allman. The prevalence of mature sac stages of T. bryosalmonae was very low. Our survey data together with knowledge of bryozoan life cycles and the ecology of PKD allow inferences concerning unknown aspects of the life cycle of T. bryosalmonae.

Notes: Proliferative kidney disease (PKD) is an economically significant disease caused by the myxozoan parasite Tetracapsula bryosalmonae. Polymerase chain reaction (PCR) protocols using primers specific for the small subunit ribosomal RNA (18S rDNA) gene of the parasite enable detection, however, false positive and negative results can render detection inconclusive. In this study a decontamination protocol was developed, using hydroxylamine hydrochloride (H), to prevent false positives by blocking re-amplification of carry-over contaminants. A mimic molecule was also developed and used as a competitive internal standard coamplified with target DNA in PCRs, revealing both true and false negatives. The sensitivity of one new and two existing primer sets was assessed with all primers detecting DNA equivalent to at least eight parasite cells per gram of tissue. This improved PCR protocol canprovide more reliable testing for T. bryosalmonae.

Notes: To better understand the possible role of deoxygenation of haemoglobin in the pathogenesis of exophthalmos in the West Australian dhufish, Glaucosoma hebraicum Richardson, some oxygen transport properties of haemoglobin from this species were determined and compared with haemoglobin from black bream, Acanthopagrus butcheri (Munro), snapper, Pagrus auratus (Bloch & Schneider) and King George whiting, Sillaginodes punctata (Cuvier). Dhufish had a single haemoglobin with a pronounced Root effect. Snapper and black bream blood had six and five types of haemoglobin, respectively. The magnitude of the Root effect of haemoglobins in the haemolysate was demonstrated by comparing oxygenation and total deoxygenation at pH 8 with relative deoxygenation at pH 6.9. Dhufish haemoglobin demonstrated a large Root effect, with the Root effect of King George whiting, snapper and black bream being of lesser magnitude. Deoxygenation of all haemolysates was more pronounced in the presence of adenosine triphosphate (ATP). Seasonal changes in haematological parameters and ATP content of dhufish blood were not evident. The Root effect is discussed in the context of its possible role in oxygen supply to the retina and adaptation to the fish's habitat and the environment.

Notes: In order to clarify the effectiveness of ultrasonication on vaccine delivery, juvenile sea bream, Pagrus major, were treated with eight different ultrasonic methods. A mixed vaccine against Vibrio alginolyticus and V. anguillarum was used to immunize the fish. The intensity and frequency of the ultrasound were 280 mW cm–2 and 35 kHz, respectively. The ultrasonic methods included continuous or pulsed ultrasound for 3 min, and continuous or pulsed ultrasound for 3 min before and/or after immersion for 3 min. Of all the eight ultrasonic methods tested, `pulsed ultrasound followed by immersion' and `immersion, pulsed ultrasound, and followed by immersion again' provided the best protection, which were comparable with protection of fish immunized by intraperitoneal injection. Moreover, the convenience of applying these two ultrasonic methods for immunization was comparable with the immersion method and was much better than intraperitoneal injection. If 2 × 108 CFU mL–1 of this mixed vaccine was used for vaccination repeatedly five times by ultrasonic methods, it could still produce good protection for the immunized sea bream. Therefore, the ultrasonic method is an effective and practical approach for fish vaccination on a large scale.

Notes: Hatchery-reared Atlantic salmon, Salmo salar L., were vaccinated intraperitoneally (i.p.) with a live attenuated Cryptobia salmositica vaccine (either 100 000 or 5000 parasites fish−1) and 4 weeks later were challenged with the parasite (either 100 000 or 5000 parasites fish−1). Unvaccinated, infected salmon had high parasitaemias and were anaemic. Fish given a high dose (100 000 parasites fish−1) had higher parasitaemias than fish given the lower dose. Vaccinated fish had low parasitaemias and a mild anaemia, but recovered quickly after challenge. Complement-fixing antibody increased in vaccinated fish after challenge and was highest at 2 weeks post-challenge. The cell-mediated response (both T cells and B cells) was depressed in infected fish until 4 weeks after infection. In vaccinated fish, the humoral response (i.e. B-lymphocytes) was greater than the cell-mediated response (i.e. T-lymphocytes). In contrast, infected fish had a greater cell-mediated than humoral immune response.

Notes: This study investigated antibody mediated immune response against Ichthyophthirius multifiliis (Ich) by determining whether theronts would retain the potential for reinfection, both in vitro and in vivo, after treatment with the culture fluid of excised skin from channel catfish, Ictalurus punctatus, immune to Ich. The invasion was reduced significantly (P 〈 0.05) for theronts treated with the immune culture fluid compared with those treated with the culture fluid from naive fish. The treatment of theronts with the immune culture fluid greatly reduced the size and survival of trophonts compared with those treated with the culture fluid from naive fish. Fewer fish were infected and the infection density was less for fish exposed to theronts treated with immune culture fluid. The infection was severe for fish invaded by theronts treated with the culture fluid from naive fish, with a high number of infected fish and heavy density of trophonts per fish. All fish were infected by Ich when exposed to the theronts treated with the immunoadsorbed culture fluid. In summary, results of this study show that cutaneous antibodies in the culture fluid of excised skin from immune fish significantly reduces theront infectivity by immobilizing or weakening theronts.

Notes: Fish from cages with histories of heavy and light amoebic gill disease (AGD) outbreaks were harvested and the morphology, histology and activities of lactate dehydrogenase determined. Although fish with a history of heavy AGD were smaller, their heart somatic indices were similar to those of fish with a history of light AGD. However, morphometrically the ratios of ventricle axis length and width and axis length and height were significantly higher, and there was an overall thickening of the muscularis compactum in the ventricle of fish with heavy AGD history. There was no difference in the lactate dehydrogenase activity of the ventricle muscle in the two fish groups. These results suggest that the change in ventricle shape associated with AGD was a possible compensation for an increased afterload where the lengthening of the ventricle was compensated for by an increase in muscle thickness, but without any overall ventricular hypertrophy or gain in ventricular mass. This suggests that AGD may be associated with cardiovascular compromise in affected fish.

Notes: Pacific herring, Clupea pallasi Valenciennes, collected from three areas of coastal British Columbia were screened for Ichthyophonus by histological examination. The infectivity of Ichthyophonus to juvenile chinook salmon, Oncorhynchus tshawytscha (Walbaum), was examined in laboratory studies. Ichthyophonus was detected in a total of 82 of 356 herring from all three areas. Prevalence in 2000 and 2001 ranged from 10.5 to 52.5% and was significantly lower in more northern (Hecate Strait) samples. Ichthyophonus was detected by histological examination in chinook salmon following oral or intraperitoneal (i.p.) exposure to homogenates of infected herring tissue. Infections in Yukon stock chinook salmon were occasionally associated with mortality and with inflammation in all tissues examined. Infections were detected significantly more frequently in the caecal mesenteries of i.p.-infected compared with oral-infected chinook salmon. The distribution and prevalence of Ichthyophonus isolates among diverse host species may assist in stock identification and in an improved understanding of trophic interactions.

Notes: Aerobic heterotrophic bacteria were isolated from the intestinal contents of Atlantic salmon, Salmo salar, rainbow trout, Oncorhynchus mykiss, and turbot, Scophthalmus maximus, on tryptone soya agar and De Man Rogosa and Sharpe agar, of which 11 of 177 (6% of the total) of the isolates were antagonistic to Aeromonas salmonicida. Four of these cultures, which were identified tentatively as A. hydrophila, Vibrio fluvialis, Carnobacterium sp. and an unidentified Gram-positive coccus, were beneficial to fish when fed singly or as an equi-mixture. Feed supplemented with the putative probiotics indicated survival of the organisms in the gastrointestinal tract for 7 days. Feeding with the probiotics for 7 and 14 days led to better survival following challenge with A. salmonicida. There was no indication of serum or mucus antibodies to A. salmonicida, but there was an increased number of erythrocytes, macrophages, lymphocytes and leucocytes, and enhanced lysozyme activity in the fish.

Notes: Edwardsiella ictaluri is a primary bacterial pathogen of channel catfish, Ictalurus punctatus, and the causative agent of enteric septicaemia of catfish. Edwardsiella ictaluri is known to gain entry to the host by infection of the nares, gastrointestinal tract, and gills, and to disseminate to organs via an as yet uncharacterized acute bacteraemia. In this study, fluorescent microscopy showed E. ictaluri on the gill within 5 min of immersion challenge and E. ictaluri could also be isolated from the circulation within 5 min. When removed to clean water, catfish cleared circulating bacteria within 15 min and the blood remained free of E. ictaluri until its reappearance at the 12 h post-infection sampling. However, Aeromonas hydrophila, the aetiological agent of motile aeromonad septicaemia, appeared within the circulation 7 h post-challenge with E. ictaluri and was detected in all fish at 12 h post-infection. Only 20% of fish carried A. hydrophila in the trunk kidney that could be detected by plate culture on Rimler–Shotts agar; however, 100% of challenged and stress-control fish were A. hydrophila complex positive at 24 h post-challenge. These results suggest that although the catfish is capable of clearing its circulation of E. ictaluri, superinfection with latent A. hydrophila may enhance clinical signs of edwardsiellosis. This is the first report of a bacterial superinfection appearing in fish.

Notes: Serum immunoglobulin, lysozyme and classical and alternative complement activity were analysed in different groups of wild and captive southern bluefin tuna (SBT), Thunnus maccoyii (Castelnau), from ambient water temperatures of 12 ± 1 and 20 ± 1 °C. Groups held captive for the longest time were found to have the highest levels of these humoral immune mediators, despite a drop in ambient temperature from 20 ± 1 to 12 ± 1 °C during the captivity period. Therefore, it may be that the immune response in these endothermic fish is not inhibited by low temperature to the extent seen in poikilothermic fish. Also, length of time in captivity appears to be associated with increased antigen exposure to maintain high levels of humoral immune mediators in these groups. Lysozyme activity was optimal at pH 5.8 and 6.2, suggesting that two isoforms, with different pH optima, are present. The SBT serum was found to lyse sheep erythrocytes by both classical and alternative complement pathways. Classical pathway activity occurred in the absence of prior sensitization with antiserum to sheep red blood cells, suggesting that natural antibodies may be present (or lectin or C-reactive protein mediated activation). Complement activity was relatively resistant to freezing at −20 °C but heating at between 45 and 50 °C for 20 min destroyed all complement activity.

Notes: Largemouth bass virus (LMBV) is a recently discovered iridovirus that causes a fatal disease of largemouth bass, Micropterus salmoides (Lacepède). Fish can become infected by waterborne LMBV, but oral transmission of this virus has not been demonstrated previously. Largemouth bass were gavaged with guppies, Poecilia reticulata Peters, which had been injected with LMBV, and then sampled periodically during a 7-week observation period. The dose of LMBV averaged 105.6 tissue culture infectious doses – 50% cytopathic endpoint (TCID50) per largemouth bass. Five of 24 largemouth bass exposed to LMBV became infected with the virus, but none of the fish had clinical signs typical of LMBV disease. Virus titres in largemouth bass were highest in swim bladder (105.5–9.5 TCID50 g−1) and were 105.2 TCID50 g−1 or lower in cutaneous mucus, head kidney, trunk kidney, spleen, gonad and intestine. These results indicate that LMBV can be transmitted orally to largemouth bass, but further study is needed to determine the factors affecting pathogenicity of the virus.

Notes: Amoebic gill disease (AGD), caused by the protozoan Neoparamoeba pemaquidensis (Page, 1987) is the most important disease affecting salmon farms in Tasmania. Reservoirs for this protozoan parasite are largely unknown. This study investigated wild fish as a potential reservoir of N. pemaquidensis. A total of 325 wild fish, comprising 12 different fish species, were caught from and around salmon farms and examined for the presence of AGD. None of the wild fish were infected with AGD. In a laboratory trial, seahorse, Hippocampus abdominalis, greenback flounder, Rhombosolea tapirina, and Atlantic salmon, Salmo salar, were challenged with N. pemaquidensis. Neoparamoeba pemaquidensis was detected on the gills on 10 of 15 (66.7%) flounder, nine of 24 (37.5%) seahorses, and six of six (100%) Atlantic salmon. However, paramoebae positive flounder and seahorse lacked the characteristic AGD gill pathology. It is concluded that AGD does not appear in wild fish and wild fish do not seem to be a reservoir of the pathogen.