The effects of sodium bicarbonate ingestion on swimming interval performance in trained competitive swimmers.

The use of sodium bicarbonate (NaHCO3) supplementation to improve repeated high-intensity performance is recommended; however, most swimming performance studies examine time trial efforts rather than repeated swims with interspersed recovery that are more indicative of training sessions. The aim of this study, therefore, was to investigate the effects of 0.3 g.kg-1 BM NaHCO3 supplementation on sprint interval swimming (8 × 50 m) in regionally trained swimmers. Fourteen regionally competitive male swimmers (body mass (BM): 73 ± 8 kg) volunteered for this double-blind, randomised, crossover designed study. Each participant was asked to swim 8 × 50 m (front crawl) at a maximum intensity from a diving block, interspersed with 50 m active recovery swimming. After one familiarisation trial, this was repeated on two separate occasions whereby participants ingested either 0.3 g.kg-1 BM NaHCO3 or 0.05 g.kg-1 BM sodium chloride (placebo) in solution 60 min prior to exercise. Whilst there were no differences in time to complete between sprints 1-4 (p > 0.05), improvements were observed in sprint 5 (p = 0.011; ES = 0.26), 6 (p = 0.014; ES = 0.39), 7 (p = 0.005; ES = 0.60), and 8 (p = 0.004; ES = 0.79). Following NaHCO3 supplementation, pH was greater at 60 min (p < 0.001; ES = 3.09), whilst HCO3- was greater at 60 min (p < 0.001; ES = 3.23) and post-exercise (p = 0.016; ES = 0.53) compared to placebo. These findings suggest NaHCO3 supplementation can improve the latter stages of sprint interval swimming performance, which is likely due to the augmentation of pH and HCO3- prior to exercise and the subsequent increase in buffering capacity during exercise.

High dose Nitrate ingestion does not improve 40 km cycling time trial performance in trained cyclists.

This study evaluated the chronic effects of nitrate (NO3-) ingestion over three days, on 40 km TT performance in 11trained cyclists (VO2max: 60.8 ± 7.4 ml.kg-1.min-1; age: 36 ± 9 years; height: 1.80 ± 0.06 m; body mass: 87.2 ± 12.0 kg). Utilising a double-blind randomised cross-over design, participants completed three 40 km TT on a Velotron® ergometer following the ingestion of either a 140 ml of "BEET It sport®" NO3- shot containing 12.8 mmol or 800 mg of NO3-, a placebo drink or nothing (control). Performance, oxygen consumption (VO2), blood bicarbonate (HCO3-), pH and lactate (BLa) and ratings of perceived exertion (RPE) were measured every 10 km throughout the TT. The present findings show that NO3- ingestion had no effect on TT performance (NO3-: 4098.0 ± 209.8 vs. Placebo: 4161.9 ± 263.3 s, p = 0.296, ES = 0.11), or VO2 (p = 0.253, ES = 0.13). Similarly, blood lactate and RPE were also unaffected by the experimental conditions (p = 0.522, ES = 0.06; p = 0.085, ES = 0.30) respectively. Therefore, these results suggest that a high dose of NO3- over three days has limited efficacy as an ergogenic aid for 40 km TT cycling performance in trained cyclists.

Biogeochemical plant-soil interaction: variable element composition in leaves of four plant species collected along a south-north transect at the southern tip of Norway.

Leaves from four different plant species (birch, willow, juniper, and heather) together with samples of the soil O and C horizons were collected at 44-46 sites along a south-to-north transect extending inland for 200 km from the southern tip of Norway. The transect covers one of the steepest vegetation gradients on Earth, crossing six vegetation zones. Juniper and heather are evergreen, and preferably exclude potentially toxic elements to avoid their accumulation in assimilating tissues, birch and willow shed their leaves in autumn together with the load of potentially toxic elements, and thus can tolerate the uptake of such elements. The plant leaves show the highest concentrations for B, Ca, K, Mg, Mn, P, Rb and S. In the soil O-horizon Ag, Au, As, Bi, Cu, Ge, Hg, In, Pb, Sb, Se, Sn, Te and W are enriched with respect to the C-horizon, whilst Mn and Rb are depleted. Cadmium, Sr and Zn are enriched in willow and Cs, Na and Tl in heather. In terms of concentration gradients from the coast inland, two different patterns are detected: 1) short range with an almost exponential decrease of concentrations from the coast, which appears to be typical for seaspray-related element input, and 2) long range with an almost linear decrease of concentrations with distance from the coast. These patterns differ among the four species, even for one and the same element. Inter-element correlation is different from material to material. Along the transect each of the different plants at the same site individually adapts to the available element combination. High linear correlations in the plants occur between the lanthanides (La, Ce, Y), and interestingly, between P and Ti. The plant/soil system appears highly non-linear and self-regulated.

Estimating the need for dental sedation. 4. Using IOSN as a referral tool.

OBJECTIVE: In this article we report on the use of the IOSN as a referral tool in primary care and the need for sedation in the referred patient population (as determined by the IOSN score). SETTING: Four centres in the North West of England (primary care) accepting referrals for treatment with the aid of sedation participated in this study. DESIGN: A service evaluation. SUBJECTS (MATERIALS) AND METHODS: The four were provided with IOSN referral forms, operator and patient questionnaires. The centres distributed IOSN forms to referrers as a means of recommending patients for sedation. All patients receiving treatment under sedation (having been referred for treatment through the IOSN form) were asked to complete the patient questionnaire. The individual operator who undertook the treatment under sedation was asked to complete the operator questionnaire. Data were entered into SPSS and the IOSN score noted. Statistical analyses of the data utilised descriptives and comparisons between groups using the Chi Squared test. RESULTS: Seventy-eight percent of the patients (n = 140) in this study were receiving treatment with sedation appropriately according to the principals of the IOSN. Patients deemed by the IOSN tool to have a low need for sedation were less likely to cancel their appointment if sedation had not been given. The majority of patients were female (70%) and the majority of operators and patients reported the IOSN forms acceptable for use. CONCLUSIONS: This study provides support for using the IOSN as a tool for organising sedation referral. The majority of operators and patients reported the IOSN forms acceptable for use.

Estimating the need for dental sedation. 1. The Indicator of Sedation Need (IOSN) - a novel assessment tool.

While the control of pain and anxiety is fundamental to the practice of dentistry, the use of conscious sedation in dentistry is very variable among dentists. The need for conscious sedation could be considered by assessing and ranking a combination of information on patient anxiety, medical history and the complexity of the anticipated clinical treatment. By undertaking this systemtic assessment an indication of sedation need may be developed which would act as an aide to decision making and, potentially, referral management. Such a tool could also be used by commissioners who need to identify patients who need conscious sedation for dental treatment in order to plan, commission and deliver appropriate sedation services.

Estimating the need for dental sedation. 2. Using IOSN as a health needs assessment tool.

AIM: This service evaluation assessed the need for sedation in a population of dental attenders (n = 607) in the North West of England. METHODS: Using the novel IOSN tool, three clinical domains of sedation need were assessed: treatment complexity, medical and behavioural indicators and patient reported anxiety using the Modified Dental Anxiety Scale. RESULTS: The findings suggest that 5% of the population are likely to require a course of treatment under sedation at some time. All three clinical domains contributed to the IOSN score and indication of treatment need. Females were 3.8 times more likely than males to be placed within the high need for sedation group. Factors such as age, deprivation and practice location were not associated with the need for sedation. CONCLUSIONS: Primary care trusts (PCTs) need health needs assessment data in order to commission effectively and in line with World Class Commissioning guidelines. This study provides both an indicative figure of need as well as a tool by which individual PCTs can undertake local health needs assessment work. Caution should be taken with the figure as a total need within a population as the study has only included those patients that attended dental practices.

Proteolytic activity of the house dust mite allergen Der p 1 enhances allergenicity in a mouse inhalation model.

BACKGROUND: We have recently demonstrated that intraperitoneal immunization of mice with proteolytically active Der p 1, the major house dust mite allergen, results in a significant and selective enhancement of total and Der p 1-specific IgE synthesis compared to mice immunized with proteolytically inactive Der p 1. OBJECTIVE: To investigate whether the proteolytic activity of Der p 1 would lead to enhanced inflammatory cellular infiltration of the lungs and systemic IgE production when administered through the respiratory system, which is the natural route of entry for this allergen. METHODS: Groups of mice were initially sensitized with proteolytically active Der p 1 through the intraperitoneal and the subcutaneous routes and subsequently exposed intranasally to either proteolytically active Der p 1, inactive Der p 1 or PBS. The extent of cellular infiltration of the lungs and systemic IgE production in the three animal groups were then compared. RESULTS: Here, we show for the first time that the administration of proteolytically active Der p 1 to mice through the intranasal route leads to significant inflammatory cellular infiltration of the lungs and systemic production of IgE. CONCLUSIONS: These data underline the important role of the proteolytic activity of Der p 1 in driving the allergic response in the lungs.

The proteolytic activity of the major dust mite allergen Der p 1 conditions dendritic cells to produce less interleukin-12: allergen-induced Th2 bias determined at the dendritic cell level.

BACKGROUND: The proteolytic activity of the house dust mite allergen Der p 1 has recently been shown to bias Th cell subset development in favour of Th2. Apart from its direct effect on T cells, it is conceivable that the proteolytic activity of Der p 1 may induce the generation of dendritic cells (DCs) that favour a Th2 response. OBJECTIVE: To study the effect of the proteolytic activity of Der p 1 on DC functions; namely cell surface phenotype, IL-12 production and ability to favour a Th2 response. METHODS: We have generated immature DCs from peripheral blood monocytes, matured them with LPS in the presence of either proteolytically active or inactive Der p 1 and compared their functions using flow cytometric analysis. RESULTS: Here we demonstrate for the first time that DCs that have been matured in the presence of proteolytically active Der p 1 produce significantly less IL-12, compared to DCs that have been matured in the presence of proteolytically inactive Der p 1. The suppression of IL-12 production was due to the cleavage of CD40 by the proteolytic activity of Der p 1, hence rendering the DCs less responsive to stimulation through the CD40L-CD40 pathway. Furthermore, we demonstrate that DCs that have been matured in the presence of proteolytically active Der p 1 induce the production of significantly less IFN-gamma and more IL-4 by CD4 T cells, compared to DCs that have been matured in the presence of proteolytically inactive Der p 1. CONCLUSIONS: Collectively, our data provide compelling evidence for the role of the proteolytic activity of Der p 1 in directing DCs to induce Th2 subset development.

The production and characterisation of a chimaeric human IgE antibody, recognising the major mite allergen Der p 1, and its chimaeric human IgG1 anti-idiotype.

BACKGROUND: Two mouse monoclonal antibodies have been described, namely: mAb 2C7 (IgG2bkappa), which is directed against the major house dust mite allergen Der p 1, and mAb 2G10 (IgG1kappa), which is an anti-idiotypic antibody raised against mAb 2C7. Given its broad IgE specificity, anti-idiotype mAb 2G10 could potentially have immunomodulatory applications. For example, a chimaeric human IgG version of mAb 2G10 could prove to be a useful molecule for binding to mast cell and basophil FcepsilonRI bound IgE, and in doing so co-ligating FcepsilonRI with FcgammaRIIB, which has been reported to have downregulatory effects. AIMS: To produce a chimaeric human IgE version of mAb 2C7 (mAb 2C7huE) and a chimaeric human IgG1 version of its anti-idiotype mAb 2G10 (mAb 2G10huG1). METHODS: The Vkappa and VH regions of mAb 2C7 and its anti-idiotype mAb 2G10 were engineered into human constant regions of the IgE and IgG1 isotypes, respectively. RESULTS: The production of chimaeric mAb 2C7huE and its anti-idiotype mAb 2G10huG1 confirmed that the respective mouse antibody V regions were successfully engineered into human constant regions and still retained the specificity of the original murine V regions. CONCLUSION: The newly constructed chimaeric antibodies will be useful to investigate the downregulation of IgE mediated hypersensitivity by the crosslinking of FcepsilonRI with FcgammaRIIB.

The proteolytic activity of the major dust mite allergen Der p 1 enhances the IgE antibody response to a bystander antigen.

BACKGROUND: We have recently demonstrated that immunization of mice with proteolytically active Der p 1, the major dust mite allergen, results in a significant enhancement in total and Der p 1-specific IgE synthesis compared to mice immunized with Der p 1 that has been irreversibly blocked with the cysteine protease inhibitors E-64 and iodoacetamide. Thus, the demonstration that the proteolytic activity of Der p 1 enhances total IgE production, apart from increasing Der p 1-specific IgE, suggests that this allergen may have an IgE-specific adjuvant effect. OBJECTIVE: To determine if the proteolytic activity of Der p 1 has an IgE-specific adjuvant effect. METHODS: We have examined this concept in experiments whereby ovalbumin, used as a bystander antigen, was injected alone or coinjected with either proteolytically active or inactive Der p 1 into groups of mice and IgE and IgG antibody responses were measured. RESULTS: Here we demonstrate for the first time that the proteolytic activity of Der p 1, when given at 10-fold higher concentration, enhances the IgE antibody response to ovalbumin. CONCLUSIONS: These findings show that the proteolytic activity of Der p 1 leads to the augmentation of IgE antibody responses to itself and to other allergens present in the microenvironment.

Human T cell subset commitment determined by the intrinsic property of antigen: the proteolytic activity of the major mite allergen Der p 1 conditions T cells to produce more IL-4 and less IFN-gamma.

The house dust mite Dermatophagoides pteronyssinus allergen Der p 1 elicits IgE antibody responses in a significant proportion of patients suffering from dust mite allergy. We have recently shown that Der p 1 proteolytically cleaves a cell surface molecule involved in the homeostatic control of human IgE synthesis, namely the IL-2 receptor (CD25) on T cells. As a result, these T cells show markedly diminished proliferation and IFN-gamma secretion in response to stimulation by anti-CD3 antibody. However, these observations still leave open the important issue of whether CD25 cleavage, and the consequent suppression of IFN-gamma secretion, leads to enhanced IL-4 secretion, and whether such cytokine changes would be exhibited by both CD4 and CD8 T cells. Here we demonstrate for the first time that the proteolytic activity of Der p 1 biases human CD4 and CD8 T cells towards a type 2 cytokine profile. Our data provide compelling evidence for the role of the proteolytic activity of Der p 1 in creating a microenvironment conducive for IgE synthesis.

Is clinical pathology accreditation worth it? A survey of CPA-accredited laboratories.

Following two pilot studies, Clinical Pathology Accreditation (CPA) accreditation was introduced to UK pathology laboratories in 1992. Since then, significant numbers of laboratories have undergone accreditation but many have never applied. We carried out a postal survey of 145 accredited laboratories in the UK to independently determine the opinions of laboratory managers/clinicians about CPA and whether accreditation had produced any significant benefits to pathology services. Ninety-three replies were received (64 per cent) a good response to an unsolicited questionnaire. Most laboratories felt accreditation by CPA had resulted in better laboratory performance with more documentation and better health and safety and training procedures. CPA accreditation was believed to provide useful information by approximately 50 per cent of laboratories but was also felt by a significant proportion of laboratories to be over-bureaucratic, inefficient and expensive (46 of 93 respondents). Many complaints were voiced about the excessive paperwork that CPA generated and there was also a significant body of opinion that felt that CPA assessed areas were the domain of other regulatory bodies such as the CPSM, IBMS and HSE.

The cysteine protease activity of the major dust mite allergen Der p 1 selectively enhances the immunoglobulin E antibody response.

The house dust mite Dermatophagoides pteronyssinus allergen Der p 1 is the most immunodominant allergen involved in the expression of dust mite-specific immunoglobulin (Ig)E-mediated hypersensitivity. The reason for this potent IgE-eliciting property of Der p 1 remains unknown, but there is mounting in vitro evidence linking the allergenicity of Der p 1 to its cysteine protease activity. Here we demonstrate for the first time that immunization of mice with proteolytically active Der p 1 results in a significant enhancement in total IgE and Der p 1-specific IgE synthesis compared with animals immunized with Der p 1 that was irreversibly blocked with the cysteine protease inhibitor E-64. We conclude that the proteolytic activity of Der p 1 is a major contributor to its allergenicity.

Activation of follicle cell surface phospholipase by tyrosine kinase dependent pathway is an essential event in Ascidian fertilization.

Eggs of Ascidia ceratodes and Phallusia mammillata block polyspermy by releasing a phosphatidylinositol-linked glycosidase from the follicle cell and egg surface that binds to and blocks all unoccupied sperm binding sites on the vitelline coat. Release of this glycosidase is thought to be under the control of a membrane-bound phospholipase. To elucidate the mechanism of phospholipase activation, intact eggs and isolated follicle cells are activated by either sperm or the tyrosine kinase activator 9, 10-dimethyl-1,2-benzanthracene (DMBA). Both treatments caused release of comparable quantities of glycosidase activity, the earliest event following fertilization. A corresponding increase in phospholipase activity accompanied this glycosidase release. The tyrosine kinase inhibitor genistein blocked release by DMBA at concentrations as low as 1 microM, but had no effect on sperm-induced release even when used up to 100 microM. Tyrphostin A23, another tyrosine kinase inhibitor, when used at 200 microM blocked glycosidase release and decreased phospholipase activity following both DMBA activation and fertilization. Western blot analysis probing for phosphotyrosine content of disrupted intact eggs with their follicle cells revealed the absence of a band in tyrphostin-treated eggs corresponding to a 40 kDa protein that was present in both unfertilized and fertilized egg samples. Based on these results, we propose that phosphorylation of specific tyrosine residues is necessary for phospholipase activation and is sufficient to trigger subsequent glycosidase release.

Evidence for cell surface and internal phospholipase activity in ascidian eggs.

Upon fertilization, ascidian eggs release a cell surface glycosidase used in the block to polyspermy and undergo cortical contractions resulting from increased intracellular calcium levels. The glycosidase is released by fertilization, calcium ionophores or added phospholipase C (PLC) activity. The PLC inhibitor D609 blocks glycosidase release. Intact Ascidia ceratodes eggs cleave 4-methylumbelliferyl-phospho-choline when it is added to seawater. This yields highly fluorescent 4-methylumbelliferone. Authentic phospholipase C but not phospholipase D can cleave this substrate. Thus, the authors believe that cleavage of the substrate is specific for PLC activity. Eggs incubated in the fluorogenic substrate after having been washed and detergent extracted were not fluorescent. Therefore the substrate failed to enter intact cells. Glycosidase release and PLC activity were stimulated by ionomycin. Octylglucoside or Triton X-100 extracts of ascidian eggs had two forms of phospholipase activity as shown by ion affinity chromatography: PL1 eluting at 0.25 mol/L NaCl and PL2 eluting at 0.6 mol/L NaCl. The PL1 appeared to be isolated as a single protein. When surface proteins were labeled with non-penetrating biotin and were subsequently reacted with streptavidin, half of the PLC activity bound. This demonstrates that half the ascidian egg PLC activity is located on the surface of either the egg or follicle cell, and half is located within the egg.

Reduction of glutamine synthetase specific activity in cultured astroglia by ferrous chloride.

Immature and mature rat astroglia in culture were assayed for glutamine synthetase (GS) activity after a single exposure to the epileptogen FeCl2. Cells were cultured with both standard and elevated extracellular potassium or glutamate (Glu) concentrations. FeCl2 reduced GS activity below control levels, whereas high Glu increased GS activity. However, stimulation by high Glu was significantly attenuated in cultures given both FeCl2 and high Glu, indicating that cells treated with FeCl2 were not able to respond as effectively to increased extracellular glutamate by increasing their GS activity. The significance of these findings is that glial regulation of the neuronal environment may be impaired, based on the proposed importance of GS in ammonia detoxification in the brain.

Pharmacokinetics of subcutaneous azidothymidine in rhesus monkeys.

The pharmacokinetics of subcutaneous bolus and continuous infusion azidothymidine (AZT) was studied in rhesus monkeys. Three animals received 100 mg/m2 as a bolus injection both intravenously and subcutaneously, with the order of administration randomly determined. Two animals received a continuous subcutaneous infusion of 25 mg/m2 per h for 12 or 24 h. AZT was measured in plasma by a reverse-phase high-pressure liquid chromatographic assay. Following intravenous bolus administration, AZT elimination was rapid, with a mean half-life of 1.2 h and a mean clearance of 318 ml/min per m2 (range, 200 to 441 ml/min per m2). The bolus subcutaneous dose was rapidly (time to peak concentration, 15 to 30 min) and nearly completely (fraction absorbed, 92%) absorbed without evidence of local tissue toxicity. With continuous subcutaneous infusion of AZT, the steady state was attained within 4 h and steady-state concentrations in plasma in the two animals exceeded 3.0 mumol/liter. No local tissue toxicity was observed at the infusion site. The subcutaneous route may be a practical alternative to intravenous administration of AZT and deserves further clinical study.