Implementing nitrous oxide cracking technology in the labour ward to reduce occupational exposure and environmental emissions: a quality improvement study.

Nitrous oxide, a potent greenhouse gas, is a common labour analgesic. One method which may reduce its carbon footprint is to 'crack' the exhaled gas into nitrogen and oxygen using catalytic destruction. In this quality improvement project, based on environmental monitoring and staff feedback, we assessed the impact of nitrous oxide cracking technology in the maternity setting. Mean ambient nitrous oxide levels were recorded during the final 30 minutes of uncomplicated labour in 36 cases and plotted on a run chart. Interventions were implemented in four stages, comprising: stage 1, baseline (12 cases); stage 2, cracking with nitrous oxide delivered and scavenged via a mouthpiece (eight cases); stage 3, cracking with nitrous oxide via a facemask with an air-filled cushion (eight cases); stage 4, cracking with nitrous oxide via a low-profile facemask, and enhanced coaching on the use of the technology (eight cases). The median ambient nitrous oxide levels were 71% lower than baseline in stage 2 and 81% lower in stage 4. Staff feedback was generally positive, though some found the technology to be cumbersome; successful implementation relies on effective staff engagement. Our results indicate that cracking technology can reduce ambient nitrous oxide levels in the obstetric setting, with potential for reductions in environmental impacts and occupational exposure.

Substantial long-term loss of alpha and gamma diversity of lake invertebrates in a landscape exposed to a drying climate.

Many regions across the globe are shifting to more arid climates. For shallow lakes, decreasing rainfall volume and timing, changing regional wind patterns and increased evaporation rates alter water regimes so that dry periods occur more frequently and for longer. Drier conditions may affect fauna directly and indirectly through altered physicochemical conditions in lakes. Although many studies have predicted negative effects of such changes on aquatic biodiversity, empirical studies demonstrating these effects are rare. Global warming has caused severe climatic drying in southwestern Australia since the 1970s, so we aimed to determine whether lakes in this region showed impacts on lake hydroperiod, water quality, and α, ß and γ diversity of lake invertebrates from 1998 to 2011. Seventeen lakes across a range of salinities were sampled biennially in spring in the Wheatbelt and Great Southern regions of Western Australia. Multivariate analyses were used to identify changes in α, ß and γ diversity and examine patterns in physicochemical data. Salinity and average rainfall partially explained patterns in invertebrate richness and assemblage composition. Climatic drying was associated with significant declines in lake depth, increased frequency of dry periods, and reduced α and γ diversity (γ declined from ~300 to ~100 taxa from 1998 to 2011 in the 17 wetlands). In contrast, ß diversity remained consistently high, because each lake retained a distinct fauna. Mean α diversity per-lake declined both in lakes that dried and lakes that did not dry out, but lakes which retained a greater proportion of their maximum depth retained more α diversity. Accumulated losses in α diversity caused the decline in γ diversity likely through shrinking habitat area, fewer stepping stones for dispersal and loss of specific habitat types. Biodiversity loss is thus likely from lakes in drying regions globally. Management actions will need to sustain water depth in lakes to prevent biodiversity loss.

Multiple melanoma susceptibility factors function in an ultraviolet radiation response pathway in skin.

BACKGROUND: Exposure to ultraviolet radiation (UVR) and the familial melanoma susceptibility gene p16 (CDKN2A) are among the major risk factors which have been identified to contribute to the development of melanoma, and also significantly contribute to squamous cell carcinoma. We have previously shown that UVR induces p16(CDKN2A) expression in melanoma and keratinocyte cell lines and human skin, but the regulatory mechanisms controlling this expression are unknown. OBJECTIVES: To determine the mechanism by which UVR induces p16(CDKN2A) expression in melanocytes and keratinocytes in the epidermis. METHODS: We have used an in vitro cell lines model of the UVR response in skin to assess the changes in p16(CDKN2A) expression and the signalling pathways regulating these changes, and validated these findings in whole human skin cultures. RESULTS: We show that UVR-induced ERK signalling, mediated by BRAF, regulates p16(CDKN2A) expression at the transcriptional, and possibly translational level. CONCLUSIONS: This study demonstrates the biological connection between the known melanoma genes p16 (CDKN2A) and BRAF in a normal physiological response to UVR in the skin, and highlights the importance of defects in this biological pathway to melanoma and squamous cell carcinoma development.

Dose response to intrathecal diamorphine for elective caesarean section and compliance with a national audit standard.

BACKGROUND: This double-blind randomised controlled trial investigated the most appropriate dose of intrathecal diamorphine to use with high-dose diclofenac as part of a multimodal analgesic regimen for caesarean section under subarachnoid block. We also wished to establish whether it was possible to satisfy the Royal College of Anaesthetists postoperative pain audit recommendation for this patient group. METHODS: One hundred and twenty patients presenting for elective caesarean section under subarachnoid block were recruited and divided into four groups. Treatment was standard except that patients were given either placebo or one of three different doses of intrathecal diamorphine (100 microg, 200 microg or 300 microg). All patients were given regular paracetamol, high-dose diclofenac and an hourly subcutaneous diamorphine regimen for breakthrough pain. RESULTS: There was a dose-dependent improvement in analgesia with intrathecal diamorphine. Only 37.9% of patients given 300 microg of intrathecal diamorphine had a visual analogue pain score of 3/10 or less throughout the study. There was a dose-dependent increase in the incidence of itching with intrathecal diamorphine although the incidence of nausea and vomiting was similar between groups. CONCLUSIONS: We found that for elective caesarean section under subarachnoid block with high dose diclofenac, analgesia was optimal with 300 microg of intrathecal diamorphine. Even the highest dose of intrathecal diamorphine did not achieve the Royal College of Anaesthetists postoperative audit target that 90% of patients should have a pain score of no more than 3/10. We believe that this target is too arduous.

n-3 polyunsaturated fatty acids inhibit the antigen-presenting function of human monocytes.

Diets rich in n-3 polyunsaturated fatty acids (PUFAs) are associated with suppression of cell-mediated immune responses, but the mechanisms are unclear. We hypothesized that n-3 PUFAs can inhibit the function of human antigen-presenting cells. A prerequisite for this role of blood monocytes is the cell surface expression of major histocompatibility complex (MHC) class II molecules [human leukocyte antigen (HLA)-DR, -DP, and -DQ], aided by the presence of intercellular adhesion molecule-1 (ICAM-1) and leukocyte function associated antigens 1 and 3. We showed previously that the n-3 PUFA eicosapentaenoic acid (EPA) inhibits the expression of HLA-DR on unstimulated human monocytes in vitro, but that docosahexaenoic acid (DHA) enhances its expression. However, both n-3 PUFAs suppress the expression of HLA-DR, HLA-DP, and ICAM-1 on interferon-gamma-activated monocytes. We also established that dietary fish-oil supplementation can inhibit the expression of these surface molecules on circulating human monocytes. We subsequently showed that when EPA and DHA were combined in the same ratio as is commonly found in fish-oil-supplement capsules (3:2), there was no significant effect in vitro on the expression of HLA-DR on unstimulated monocytes, but the expression on activated monocytes remained significantly inhibited. In the same in vitro system, the ability of activated monocytes to present antigen to autologous lymphocytes was significantly reduced after culture with the combined n-3 PUFAs. These findings provide one potential mechanism for the beneficial effect of fish oil in the treatment of rheumatoid arthritis, a disorder associated with elevated expression of MHC class II and adhesion molecules on monocytes present within affected joints.

Does tomato consumption effectively increase the resistance of lymphocyte DNA to oxidative damage?

BACKGROUND: Lycopene, the main carotenoid in tomato, has been shown to be a potent antioxidant in vitro. However, there is no significant evidence of its antioxidant action in vivo. OBJECTIVE: We evaluated the effect of tomato intake on plasma carotenoid concentrations and lymphocyte resistance to oxidative stress. DESIGN: Ten healthy women (divided into 2 groups of 5 subjects each) ate a diet containing tomato puree (providing 16.5 mg lycopene) and a tomato-free diet for 21 d each in a crossover design. Before and after each diet period, plasma carotenoid concentrations and primary lymphocyte resistance to oxidative stress (evaluated by means of single-cell gel electrophoresis) were analyzed. RESULTS: After the first 21-d experimental period, total plasma lycopene concentrations increased by 0.5 micromol/L (95% CI: 0.14, 0.87) in the group that consumed the tomato diet and decreased by 0.2 micromol/L (95% CI: -0.11, -0.30) in the group that consumed the tomato-free diet (P < 0.001). Tomato consumption also had an effect on cellular antioxidant capacity: lymphocyte DNA damage after ex vivo treatment with hydrogen peroxide decreased by 33% (95% CI: 0.8%, 61%; P < 0.05) and by 42% (95% CI: 5.1%, 78%; P < 0.05) in the 2 groups of subjects after consumption of the tomato diet. CONCLUSION: The consumption of tomato products may reduce the susceptibility of lymphocyte DNA to oxidative damage.

Fish oil supplementation inhibits the expression of major histocompatibility complex class II molecules and adhesion molecules on human monocytes.

To test the hypothesis that fish oil supplementation can inhibit the expression of functionally associated molecules on the surface of human blood monocytes, we randomly assigned 12 healthy adults to receive either an n-3 polyunsaturated fatty acid-rich fish oil supplement for 21 d or to receive no supplement. The percentage of monocytes expressing major histocompatibility complex (MHC) class II molecules (HLA-DR, -DP, and -DQ), intercellular adhesion molecule-1, and leukocyte-function-associated antigen-1, and the intensity of expression of each molecule were quantified before and after the study period. Monocytes were examined immediately after blood sampling and again after incubation in serum-free culture medium for 24 h in the presence of interferon-gamma to up-regulate expression of MHC class II molecules by the monocytes. The intensity of expression of all the monocyte surface molecules examined was significantly reduced after fish oil supplementation (P < 0.025), although there was no change in the percentage of monocytes expressing each molecule. After incubation with interferon-gamma, there was a similar inhibition of surface molecule expression (with the exception of HLA-DQ) by monocytes from the fish oil-supplemented group, and there was a reduction in the percentage of monocytes expressing both HLA-DR and -DP molecules (P < 0.025). No significant changes were observed in the reference group. Dietary supplementation with fish oil can inhibit the expression of surface molecules involved in the function of human antigen-presenting cells, a potential mechanism by which n-3 fatty acids may suppress cell-mediated immune responses.

OXYGEN TRANSPORT IN EGG MASSES OF THE AMPHIBIANS RANA SYLVATICA AND AMBYSTOMA MACULATUM: CONVECTION, DIFFUSION AND OXYGEN PRODUCTION BY ALGAE

Many amphibians lay their eggs in gelatinous masses up to 10­20 cm in diameter, posing problems for diffusive oxygen delivery. Oxygen may also be provided by water convection between eggs or by oxygen production by endogenous algae. We studied egg masses of two local amphibians, Rana sylvatica and Ambystoma maculatum, to estimate the importance of each of these processes. We injected dye to check for water channels, measured oxygen partial pressures within egg masses to determine the influence of external water convection and lighting, measured oxygen consumption and production in darkness and light and calculated expected gradients through egg masses with a cylindrical, homogeneous egg mass model. Rana sylvatica had relatively loose egg masses with water channels between the eggs; water convection was important for oxygen delivery. Ambystoma maculatum had firm egg masses with no spaces in the jelly between eggs; thus, there was no opportunity for convective oxygen delivery. The egg masses were cohabited by Oophila ambystomatis, a green alga found specifically in association with amphibian egg masses. Oxygen delivery in A. maculatum was by diffusion and by local production by the algal symbiont. Analysis of a cylindrical egg mass model and measurement of oxygen gradients through egg masses indicated that diffusion alone was not adequate to deliver sufficient O2 to the innermost embryos at late developmental stages. In the light, however, egg masses had a net oxygen production and became hyperoxic. Over the course of a day with a 14 h:10 h light:dark cycle, the innermost embryos were alternately exposed to hyperoxia and near anoxia.

The respiratory development of Atlantic salmon. I. Morphometry of gills, yolk sac and body surface

During development from larva to juvenile in Atlantic salmon, Salmo salar, there is a change in the anatomical potential for gas exchange among gills, body skin and yolk sac as the larvae resorb yolk, grow and develop gills. Newly hatched Atlantic salmon have poorly developed gills but do have a high skin area to mass ratio and a large well-vascularized yolk sac. Cutaneous surfaces accounted for over 95 % of the total area available for respiration in newly hatched Atlantic salmon (body mass 0.032­0.060 g). The branchial contribution to total area increased rapidly, however, so that by the end of yolk absorption (body mass 0.19­0.23 g) it constituted 22 % of the total area and overtook cutaneous surface area between 5 and 6 g wet body mass. Harmonic mean diffusion distance across the skin increased through development from 20 µm at hatch (14 µm across the yolk sac) to 70 µm in an 11 g fish. Diffusion distances across both the filaments and lamellae of the gills decreased through development, from 3.7 to 2.4 µm for lamellae and from 14.5 to 10.8 µm for filaments. The total anatomical diffusion factor (ADF, mass-specific surface area per unit diffusion distance) remained constant over early development and appeared to be higher than in adult fish. The distribution of ADF changed over early development from 50 % yolk sac, 42 % body surface and 8 % branchial in newly hatched fish to 68 % branchial and 32 % cutaneous at the end of yolk resorption. Generally, early post-hatch development of gills, ADF and some cutaneous surfaces showed high mass exponents. After yolk resorption (body mass 0.2 g), however, these coefficients were lower and closer to unity. The change in scaling at the end of yolk resorption in this study may reflect the completion of larva to juvenile metamorphosis in Atlantic salmon. Comparison between our data and values in the literature suggests that the timing of gill development is related more to developmental stage than to body size.

The respiratory development of Atlantic salmon. II. Partitioning of oxygen uptake among gills, yolk sac and body surfaces

During post-hatch development of Atlantic salmon (Salmo salar L.), O2 uptake partitioning changes from primarily cutaneous to primarily branchial. Over 80 % of post-hatch O2 uptake was cutaneous, with the yolk sac responsible for 33 % of total O2 uptake. The well-vascularized yolk sac was a less effective gas exchanger than the unperfused skin of the body, suggesting that oxygen delivery is by direct diffusion to the tissues. Branchial O2 uptake increased quickly as gill lamellae developed, contributing 60 % of total O2 uptake before the completion of yolk resorption (body mass 0.2 g) and increasing to 69­81 % in fish weighing over 0.3 g. The area-specific O2 uptake of the skin decreased through development as skin thickness increased, while that of the gills increased from 0.10 µg h-1 mm-2 to 0.23 µg h-1 mm-2. Partitioning of O2 uptake of the skin and gills changed in concert with changes in the partitioning of the anatomical diffusion factor (ADF, mass-specific surface area per unit diffusion distance) between skin and gills, which changed from more than 95 % to less than 10 % cutaneous; thus, ADF is a useful rough indicator of oxygen uptake potential. Caution should be used in predicting oxygen uptake potential from ADF, however, because O2 uptake per unit diffusion barrier of the yolk sac was less than half that of the general body surface, and O2 uptake per unit diffusion barrier of the gills changed dramatically over development.

EFFECT OF CORONARY ABLATION AND ADRENERGIC STIMULATION ON IN VIVO CARDIAC PERFORMANCE IN TROUT (ONCORHYNCHUS MYKISS)

In fish, catecholamine-induced changes in cardiac performance in vivo are the result of complex interactions between the direct adrenergic effects on the heart and peripheral circulation and the reflex responses to increased blood pressure. In addition, coronary artery transport of catecholamines and oxygen to the compact myocardium may be essential for maximal in vivo cardiac performance during adrenergic stimulation. Cardiac output (Q(dot)), heart rate (fh), stroke volume (Vs) and dorsal aortic pressure (Pda) were measured in trout with intact or ablated coronary arteries at rest and following intra-arterial administration of 0.2, 0.5, 1.0 and 2.0 µg kg-1 adrenaline. Resting Q(dot), fh, Vs and Pda were the same in fish with intact and ablated coronaries at 48 h post-surgery, averaging approximately 18 ml min-1 kg-1, 42 beats min-1, 0.42 ml kg-1 and 2.2 kPa, respectively. All cardiovascular variables showed a strong relationship between response magnitude and adrenaline dose. However, our results indicate that adrenaline doses above 0.5 µg kg-1 may have a limited ability to increase Q(dot) (ED50 0.22 µg kg-1). Coronary artery ablation had little effect on post-injection Q(dot), Vs, Pda or fh at any dose of adrenaline. In both intact and ablated groups, two types of responses in Q(dot) were observed following adrenaline injection. In the 'type 1' response, Q(dot) increased shortly (15­30 s) after adrenaline administration, as increases in Vs more than compensated for a pressor-stimulated reflex bradycardia. In the 'type 2' response, alterations in Q(dot) were biphasic. In the initial minutes post-injection, Q(dot) fell and reached a minimum level at 1­2 min, the result of an immediate drop in fh and a delayed post-injection increase in Vs. Thereafter, Q(dot) gradually increased as a result of concordant increases in fh and Vs. Although time to maximum Q(dot) was 5­6 min longer for fish exhibiting type 2 responses, there was no difference in maximum Q(dot) increase or in the time courses for changes in fh and Pda between response types. Our results suggest (1) that during normoxic conditions, cardiac performance does not depend highly on coronary blood flow; (2) that the capacity of adrenaline to increase Q(dot) may be limited by elevations in output pressure and/or by the low dose (concentration) of adrenaline required to achieve near maximal adrenergic stimulation of the heart; and (3) that fish exhibiting type 2 responses have an increased barostatic gain (%delta fh per unit Pda) compared with those with type 1 responses.

INFLUENCE OF HYPOXIA AND ADRENALINE ADMINISTRATION ON CORONARY BLOOD FLOW AND CARDIAC PERFORMANCE IN SEAWATER RAINBOW TROUT (ONCORHYNCHUS MYKISS)

To investigate the relationship between cardiac performance and coronary perfusion, cardiovascular variables (Q(dot), Vs, fh, Pda) and coronary blood flow (q·cor) were measured in rainbow trout (Oncorhynchus mykiss) (1.2­1.6 kg) before and after adrenergic stimulation (1.0 µg kg-1 adrenaline) under conditions of (1) normoxia, (2) hypoxia (approximate PwO2 12 kPa) and (3) 2.5 h after returning to normoxia. q·cor for resting fish under normoxic conditions was 0.14±0.02 ml min-1 kg-1 (approximately 0.85 % of Q(dot)). When exposed to hypoxia, although both resting Q(dot) and q·cor increased, q·cor increased to a greater degree (Q(dot) by 17 % and q·cor by 36 %). During hypoxia, maximum adrenaline-stimulated Q(dot) was comparable to that observed for normoxic fish. However, because Q(dot) was elevated in resting hypoxic fish, the capacity of hypoxic fish to increase Q(dot) above resting levels was 50 % lower than that measured in normoxic fish. Although maximum q·cor in adrenaline-injected hypoxic trout was greater than that measured in normoxic trout, post-injection increases in q·cor (above resting levels) were not different between the two groups. Two and a half hours after hypoxic exposure, resting Q(dot) was still elevated (11 %) above normoxic levels, and the ability to increase Q(dot) when adrenergically stimulated was not fully restored. These results suggest (1) that resting q·cor in salmonids is approximately 1 % of Q(dot), (2) that increases in q·cor may be important in maintaining cardiovascular performance during hypoxic conditions, (3) that interactions between alpha-adrenergic constriction and metabolically related vasodilation of the coronary vasculature are important in determining q·cor in fish, (4) that exposure of fish to moderate environmental hypoxia reduces the scope for adrenergically mediated increases in Q(dot), and (5) that periods of recovery in excess of several hours are required before cardiovascular performance returns to pre-hypoxic levels.

Hydro-resistive measurement of dynamic lifting strength.

A device is described for measuring strength and power outputs of dynamic vertical lifts between heights of 0.4 and 2.2 m. The device is safe, robust, and easily transportable. It consists of a water-filled tube 2 m high and 200 mm internal diameter. The subject pulls vertically on a handle which is connected with flexible wire rope via a series of pulleys to a piston suspended inside the tube. The piston has holes which can be closed with bungs. The drag force is proportional to the square of the velocity. The constant of proportionality can be chosen over a more than 100-fold range and is independent of temperature. Manual force is measured using a strain gauged cantilever over which the rope passes. Rope movement is monitored with a shaft encoder. These devices are sampled synchronously by an interfaced computer. Velocity and power are derived from the measurements of displacement, time and force. The device is highly accurate. Power measurements are not significantly different on two separate days although repetitions on one day show a warming-up effect. This device allows the study of dynamic lifts ranging from slow, high force, quasi-isokinetic lifts to lifts where high velocities and accelerations occur.

Effect of eicosapentaenoic acid on the proliferation and incidence of apoptosis in the colorectal cell line HT29.

Fish oil has been shown to reduce the induction of colorectal cancer in animal models by a mechanism which may involve suppression of mitosis, increased apoptosis, or both. We used the human colonic adenocarcinoma cell line HT29 to explore the effects of the long-chain n-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) on cell proliferation and death in vitro. Cells were cultured in media containing EPA at 5, 10, and 15 microg/mL. Cell number and thymidine incorporation were used to quantify proliferation, and cell cycle effects were studied using flow cytometry. Gel electrophoresis, annexin-V binding, and morphological criteria were used to characterize apoptosis. Adherent cells and freely floating detached cells were treated as two distinct populations. In the presence of EPA at 10 and 15 microg/mL there was a marked reduction in the growth rate of adherent HT29 colonies, owing to an increased detachment of adherent cells. After treatment with 10 or 15 microg/mL EPA the proportion of adherent cells in S-phase increased, indicating either a block in late S-phase or early G2. Floating cells showed evidence of extensive DNA cleavage, but the proportion of floating cells with sub GO DNA content declined on treatment with 10 or 15 microg/mL EPA even though the number of floating cells increased. We conclude that EPA does not inhibit mitosis of adherent cells, but increases the rate at which they become detached from the substrate, probably at an early stage in the initiation of apoptosis. This mechanism may be analogous to "anoikis," or induction of apoptosis in response to loss of cell contact, and may contribute to the anticarcinogenic effects of fish oil in vivo.

Flow cytometry and its applications in veterinary medicine.

Flow cytometry is a technique for analysing and separating populations of cells (and subcellular components). The cells are generally stained with fluorescent markers (eg, fluorescent antibodies or DNA-binding dyes). Each cell is analysed individually, at high speed. Thus, assays may be performed on small samples (less than 10,000 cells). A subpopulation of interest can be separated from the remainder of the cells to a high degree of purity (up to 99 per cent). The application of flow cytometry to veterinary science is increasing and is discussed here.

System for the triaxial measurement of manual force exertions.

A triaxial force measurement system has been developed to allow the measurement of whole-body manual force exertions in the three axes of the Cartesian system. The design of the system precludes the exertion of torques, thus enabling a description of the linear forces that an individual can exert in three dimensions. Forces are measured independently for each hand, allowing the examination of triaxial forces in both one- and two-handed whole-body exertions, and the analysis of the forces in the closed chain of the upper limbs in two-handed exertions. The hardware and computer software that constitute the triaxial force measurement system are described, as are the method of calibration used and the accuracy of the system. Asymmetric postures and exertions are seen as risk factors leading to musculoskeletal disorders. Measurement and descriptions of manual strength involving asymmetric postures and lateral components of exertion will enable risk analysis of such tasks to be undertaken.