Potential role of nitric oxide in contraction-stimulated glucose uptake and mitochondrial biogenesis in skeletal muscle

• 1. The present review discusses the potential role of nitric oxide (NO) in the: (i) regulation of skeletal muscle glucose uptake during exercise; and (ii) activation of mitochondrial biogenesis after exercise.
• 2. We have shown in humans that local infusion of an NO synthase inhibitor during exercise attenuates increases in skeletal muscle glucose uptake without affecting blood flow. Recent studies from our laboratory in rodents support these findings in humans, although rodent studies from other laboratories have yielded conflicting results.
• 3. There is clear evidence that NO increases mitochondrial biogenesis in non-contracting cells and that NO influences basal skeletal muscle mitochondrial biogenesis. However, there have been few studies examining the potential role of NO in the activation of mitochondrial biogenesis following an acute bout of exercise or in response to exercise training. Early indications are that NO is not involved in regulating the increase in mitochondrial biogenesis that occurs in response to exercise.
• 4. Exercise is considered the best prevention and treatment option for diabetes, but unfortunately many people with diabetes do not or cannot exercise regularly. Alternative therapies are therefore critical to effectively manage diabetes. If skeletal muscle NO is found to play an important role in regulating glucose uptake and/or mitochondrial biogenesis, pharmaceutical agents designed to mimic these effects of exercise may improve glycaemic control.

The effects of an area-based intervention on the uptake of maternal and child health assessments in Australia : a community trial

Background
Recognition of the importance of the early years in determining health and educational attainment and promotion of the World Health Organization Health for All (HFA) principles has led to an international trend towards community-based initiatives to improve developmental outcomes among socio-economically disadvantaged children. In this study we examine whether, Best Start, an Australian area-based initiative to improve child health was effective in improving access to Maternal and Child Health (MCH) services.

Methods
The study compares access to information, parental confidence and annual 3.5 year Ages and Stages visiting rates before (2001/02) and after (2004/05) the introduction of Best Start. Access to information and parental confidence were measured in surveys of parents with 3 year old children. There were 1666 surveys in the first wave and 1838 surveys in the second wave. The analysis of visiting rates for the 3.5 year Ages and Stages visit included all eligible Victorian children. Best Start sites included 1,739 eligible children in 2001/02 and 1437 eligible children in 2004/05. The comparable figures in the rest of the state were and 45, 497 and 45, 953 respectively.

Results
There was a significant increase in attendance at the 3.5 year Ages and Stages visit in 2004/05 compared to 2001/02 in all areas. However the increase in attendance was significantly greater at Best Start sites than the rest of the state. Access to information and parental confidence improved over the course of the intervention in Best Start sites with MCH projects compared to other Best Start sites.

Conclusion
These results suggest that community-based initiatives in disadvantaged areas may improve parents' access to child health information, improve their confidence and increase MCH service use. These outcomes suggest such programmes could potentially contribute to strategies to reduce child health inequalities.

Local nitric oxide synthase inhibition reduces skeletal muscle glucose uptake but not capillary blood flow during in situ muscle contraction in rats

OBJECTIVE: We have previously shown in humans that local infusion of a nitric oxide synthase (NOS) inhibitor into the femoral artery attenuates the increase in leg glucose uptake during exercise without influencing total leg blood flow. However, rodent studies examining the effect of NOS inhibition on contraction-stimulated skeletal muscle glucose uptake have yielded contradictory results. This study examined the effect of local infusion of an NOS inhibitor on skeletal muscle glucose uptake (2-deoxyglucose) and capillary blood flow (contrast-enhanced ultrasound) during in situ contractions in rats.

RESEARCH DESIGN AND METHODS: Male hooded Wistar rats were anesthetized and one hindleg electrically stimulated to contract (2 Hz, 0.1 ms) for 30 min while the other leg rested. After 10 min, the NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) (arterial concentration of 5 µmol/l) or saline was infused into the epigastric artery of the contracting leg.

RESULTS: Local NOS inhibition had no effect on blood pressure, heart rate, or muscle contraction force. Contractions increased (P < 0.05) skeletal muscle NOS activity, and this was prevented by L-NAME infusion. NOS inhibition caused a modest significant (P < 0.05) attenuation of the increase in femoral blood flow during contractions, but importantly there was no effect on capillary recruitment. NOS inhibition attenuated (P < 0.05) the increase in contraction-stimulated skeletal muscle glucose uptake by ~35%, without affecting AMP-activated protein kinase (AMPK) activation.

CONCLUSIONS: NOS inhibition attenuated increases in skeletal muscle glucose uptake during contraction without influencing capillary recruitment, suggesting that NO is critical for part of the normal increase in skeletal muscle fiber glucose uptake during contraction.

The effect of nitric oxide donor sodium nitroprusside on glucose uptake in human primary skeletal muscle cells

Nitric oxide (NO) has been implicated as an important signaling molecule in the insulin-independent, contraction-mediated glucose uptake pathway and may represent a novel strategy for blood glucose control in patients with type 2 diabetes (T2DM). The current study sought to determine whether the NO donor, sodium nitroprusside (SNP) increases glucose uptake in primary human skeletal muscle cells (HSkMC) derived from both healthy individuals and patients with T2DM. Vastus lateralis muscle cell cultures were derived from seven males with T2DM (aged 54 ± 2 years, BMI 31.7 ± 1.2 kg/m², fasting plasma glucose 9.52 ± 0.80 mmol/L) and eight healthy individuals (aged 46 ± 2 years, BMI 27.1 ± 1.5 kg/m², fasting plasma glucose 4.69 ± 0.12 mmol/L). Cultures were treated with both therapeutic (0.2 and 2 μM) and supratherapeutic (3, 10 and 30 mM) concentrations of SNP. An additional NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP) was also examined at a concentration of 50 μM. Glucose uptake was significantly increased following both 30 and 60 min incubations with the supratherapeutic SNP treatments (P = 0.03) but not the therapeutic SNP doses (P = 0.60) or SNAP (P = 0.54). There was no difference in the response between the healthy and T2DM cell lines with any treatment or dose. The current study demonstrates that glucose uptake is elevated by supratherapeutic, but not therapeutic doses of SNP in human primary skeletal muscle cells derived from both healthy volunteers and patients with T2D. These data confirm that nitric oxide donors have potential therapeutic utility to increase glucose uptake in humans, but that SNP only achieves this in supratherapeutic doses. Further study to delineate mechanisms and the therapeutic window is warranted.

Improving the uptake of pulmonary rehabilitation in patients with COPD : qualitative study of experiences and attitudes

Background
Pulmonary rehabilitation can improve the quality of life and ability to function of patients with chronic obstructive pulmonary disease (COPD). It may also reduce hospital admission and inpatient stay with exacerbations of COPD. Some patients who are eligible for pulmonary rehabilitation may not accept an offer of it, thereby missing an opportunity to improve their health status.

Aim
To identify a strategy for improving the uptake of pulmonary rehabilitation.

Design of study
Qualitative interviews with patients.

Setting
Patients with COPD were recruited from a suburban general practice in north-east Derbyshire, UK.

Method
In-depth interviews were conducted on a purposive sample of 16 patients with COPD to assess their concerns about accepting an offer of pulmonary rehabilitation. Interviews were analysed using grounded theory.

Results
Fear of breathlessness and exercise, and the effect of pulmonary rehabilitation on coexisting medical problems were the most common concerns patients had about taking part in the rehabilitation. The possibility of reducing the sensation of breathlessness and regaining the ability to do things, such as play with their grandchildren, were motivators to participating.

Conclusion
A model is proposed where patients who feel a loss of control as their disease advances may find that pulmonary rehabilitation offers them the opportunity to regain control. Acknowledging patients' fears and framing pulmonary rehabilitation as a way of ‘regaining control’ may improve patient uptake.

Effects of leveling agent on the uptake of reactive dyes by untreated and plasma-treated wool

Atmospheric-pressure plasma treatment of wool fabric produced a significantly higher level of adsorbed fiber-reactive dye when applied at 50 °C (pH 3.0–6.0) in the absence of any organic leveling agent. In addition, color yields indicated that dye was more uniformly adsorbed by the plasma-treated fabric compared with the untreated material. When untreated fabric was dyed in the presence of a leveling agent (Albegal B), the extent and levelness of dye sorption were enhanced. These enhancements were, however, relatively small on the plasma-treated wool compared with those on untreated wool. A ‘surface’ mechanism, similar to that proposed when plasma-treated wool is dyed in the absence of leveling agent, can explain the leveling ability of Albegal B under adsorption conditions. Increasing the dyebath temperature to 90 °C resulted in dye penetration of the fibers. Under these conditions, any enhancements of dye uptake produced by the plasma treatment, as well as the use of Albegal B, were relatively small, in contrast to the behavior at 50 °C. Improvements in the uniformity of dye sorption observed at 50 °C were, however, maintained at the higher temperature. It is concluded that the inability of reactive dyes to migrate (and so promote leveling and uniformity) once they have reacted with the fiber, means that differences in the uniformity of dye sorbed at 50 °C are still apparent at equilibrium.

Effects of plasma treatment of wool on the uptake of sulfonated dyes with different hydrophobic properties

A wool fabric has been subjected to an atmospheric-pressure treatment with a helium plasma for 30 seconds. X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry confirmed removal of the covalently-bound fatty acid layer (F-layer) from the surface of the wool fibers, resulting in exposure of the underlying, hydrophilic protein material. Dye uptake experiments were carried out at 50 ºC to evaluate the effects of plasma on the rate of dye uptake by the fiber surface, as well as give an indication of the adsorption characteristics in the early stages of a typical dyeing cycle. The dyes used were typical, sulfonated wool dyes with a range of hydrophobic characteristics, as determined by their partitioning behavior between water and n-butanol. No significant effects of plasma on the rate of dye adsorption were observed with relatively hydrophobic dyes. In contrast, the relatively hydrophilic dyes were adsorbed more rapidly (and uniformly) by the plasma-treated fabric. It was concluded that adsorption of hydrophobic dyes on plasma-treated wool was influenced by hydrophobic interactions, whereas electrostatic effects predominated for dyes of more hydrophilic character. On heating the dyebath to 90 ºC in order to achieve fiber penetration, no significant effect of the plasma treatment on the extent of uptake or levelness of a relatively hydrophilic dye was observed as equilibrium conditions were approached.

Intranet and internet uptake and usage within UK building surveying practices. A user based evaluation of advantages and disadvantages

The ‘single most important technological discovery since the birth of the p.c.’, (Gates, 1996). The swift increase in Internet utilisation within businesses and surveying has changed working methods within UK based Surveying practices. User numbers expanded rapidly since the early 1990’s and estimates place a figure of 150 million internet users globally (www.Yahoo.com, 2000). Sloan and Kee Low (2000) established that ‘e-commerce’ is a revolutionary way of trading and conducting business activities. There are opportunities for surveying practices including, reducing communication time, increasing revenues from new and existing sources, enhanced client satisfaction and competitive advantage (Cronin, 1996). Others (Dixon, 1998) perceive threats, such as ‘disintermediation’, to the traditional role of the surveyor.

This research examined the levels of internet and intranet access within the UK Building and Quantity Surveying practices and corporate firms to establish how the internet has affected working methods and client requirements. The research method comprised a qualitative approach using an email and postal questionnaire survey of a random sample of practices. The questionnaire findings formed the basis of semi structured interviews conducted with senior management of large practices to gain a deeper understanding of the issues identified within the survey.

Data analysis revealed the traditional role of the surveyor as a broker of information may be under threat in the long term. Generally levels of internet access in the sample exceed that of general business levels (Dti, 2000) with the largest growth pre 1997. The perceived advantages and barriers to Intranet and Internet usage were identified by the research.

The effect of vitamin : a deficiency on glucose uptake in the rat

This thesis describes an investigation of the effects of vitamin A deficiency on gut function, The central hypothesis to be tested was that acute vitamin A deficiency affects glucose uptake from the small intestine- The hypothesis was tested using a system involving perfusion of isolated segments of the small intestine in the anaesthetized rat. The system was used to study effects on glucose uptake under steady-state conditions. In the initial part of the study, experiments were diverted towards setting up the system for measuring steady-state uptake, and determining the relative contributions of active uptake and diffusion. Phenol red was found to be a reliable non-absorbable marker for determining net water movement. Phlorizin, generally at 1 mmol/L, was used as a competitive (reversible) inhibitor of active uptake. It is difficult however to confirm complete inhibition of active uptake by phlorizin because of the limited solubility of the inhibitor. The kinetics of glucose uptake f ram intra-luminal maltose were found to be, in general, not significantly different from those applying to the uptake of glucose from an equivalent glucose solution. Maltase activity in the perfused gut segment was found to be sufficient to hydrolyse most of the maltose (80 per cent or more) in the solution being perfused, a much greater proportion than was absorbed. Glucose absorptive capacity, measured on an intestinal dry weight basis, was greatest in the duodenum and progressively less in the jejunum and ileum. The rate of water uptake f ran the gut was increased by the presence of glucose in the lumen, and was linked to glucose uptake as shown by the inhibition of water uptake by phlorizin. Uptake of glucose by solvent drag was demonstrated by showing an increased rate of glucose uptake when the rate of water uptake was increased by perfusing a solution of reduced osmotic pressure. In the experiment a low intra-luminal glucose concentration was used to preclude net uptake by diffusion and active uptake was blocked with phlorizin. This process was further investigated using streptozotocin-diabetic rats in which the diabetes establishes a hyperosomotic blood with hyperglycaemia. Uptake by solvent drag was more obvious in diabetic animals. A back-diffusion (exsorption) of glucose from the tissues to the lumen was also shown; the rate being proportional to plasma glucose concentration. Vitamin A deficiency was established in weanling rats after 6-7 weeks feeding on a diet based on wheat starch, coconut oil, and casein washed with hot ethanol, together with vitamins and minerals. The vitamin A deficiency led to classic eye signs and was reversed by the addition to the diet of retinoic acid (5 g/g diet). Vitamin A deficiency decreased intestinal mucus production (dry weight) but had no detectable effect on the histology of the villous epithelium as shown under the light microscope. Using perfusion experiments it was shown that vitamin A deficiency had no significant effect on the rate of active uptake of glucose, but that deficiency increased the rate of passive uptake.

Nutrients in Pirron Yallock Creek, southwestern Victoria : paleolimnological and management considerations

The concentrations of various forms of nitrogen and phosphorus in the main stream and selected tributaries of Pirron Yallock Creek, southwestern Victoria were examined over a two-year period. Exceedingly high levels of both nutrients were found within a particular tributary, while generally high nutrient levels were observed throughout the catchment. The speciation results indicated that dairy effluent was the primary source of nutrients to Pirron Yallock Creek. A palaeolimnological study was undertaken to determine the impact of European settlement upon waterway nutrient concentrations within the region. The palaeolimnological study focused primarily on the diatom flora preserved within the sediments of nearby Lakes Corangamite and Bullen Merri. Lack of preservation of diatoms within the sediments of Lake Corangamite resulted in no water quality inferences for this lake. The preserved diatoms within Lake Bullen Merri suggested an increase in trophic status of this lake during the last 500 years. While a change in the diatom flora of Lake Bullen Merri was evident, it was not possible to differentiate the impact of European settlement upon nutrient status of this waterway from long-term trophic status change. In light of the particularly high nutrient concentrations observed in Pirron Yallock Creek, improved nutrient management strategies are proposed for the catchment. These strategies, which consider current nutrient management activities, are predominantly focussed on the dairy industry, which occupies the majority of the catchment.

A non-radioactive method for measuring Cu uptake in HepG2 cells

At present, all data on Cu uptake and metabolism have been derived from radioactive uptake experiments. These experiments are limited by the availability of the radioactive isotopes ⁶⁴Cu or ⁶⁷Cu, and their short half-life (12.5 and 62 h, respectively). In this paper, we investigate an alternative method to study the uptake of Cu with natural isotopes in HepG2 cells, a liver cell line used extensively to study Cu metabolism. In nature, Cu occurs as two stable isotopes, ⁶³Cu and ⁶⁵Cu (⁶³Cu/⁶⁵Cu = 2.23). This ratio can be measured accurately using inductively coupled plasma mass spectrometry (ICP-MS). In initial experiments, we attempted to measure the time course of Cu uptake using ⁶⁵Cu. The change in the ⁶³Cu/⁶⁵Cu ratio, however, was too small to allow measurement of Cu uptake by the cells. To overcome this difficulty, the natural ⁶³Cu/⁶⁵Cu ratio in HepG2 cells was altered using long-term incubation with ⁶³Cu. This had a significant effect on Cu concentration in HepG2 cells, changing it from 81.9 ± 9.46 pmol μg DNA⁻¹ (week 1) to 155 ± 8.63 pmol μg DNA⁻¹ (week 2) and stabilising at 171 ± 4.82 pmol μg DNA⁻¹ (week 3). After three weeks of culture with 2 μM ⁶³Cu the ⁶³Cu/⁶⁵Cu changed from 2.18 ± 0.05 to 15.3 ± 1.01. Cu uptake was then investigated as before using ⁶⁵Cu. Uptake was linear over 60 min, temperature dependent and consistent with previous kinetics data. These observations suggest that stable isotope ICP-MS provides an alternative technique for the study of Cu uptake by HepG2 cells.

Threshold effects of glucose transporter-4 (GLUT4) deficiency on cardiac glucose uptake and development of hypertrophy

The aim of this study was to investigate the metabolic and structural consequences of a decrease in glucose transporter-4 (GLUT4) levels on the heart. The CreLoxP system was utilised to delete GLUT4 in muscle tIssue including heart. The presence of the PGK-neoR cassette in the GLUT4-Lox mice resulted in reduced expression in all tIssues to levels 15-30% of wild-type control mice. In mice expressing Cre recombinase, there was a further reduction of GLUT4 in cardiac tIssue to almost undetectable levels. Cardiac glucose uptake was measured basally and during a uglycaemic/hyperinsulinaemic clamp using 2-deoxy-[1-(14)C]glucose. Insulin-stimulated glucose uptake was normal in hearts expressing 15% of normal GLUT4 levels but markedly reduced in mice with more profound reduction in GLUT4. Cardiac enlargement occurred only when GLUT4 levels were less than 5% of normal values. In heart there is a threshold level of GLUT4 above which insulin-stimulated glucose uptake is maintained. As little as 5% of normal GLUT4 levels expressed in heart is sufficient to prevent the development of cardiac hypertrophy. 2-deoxy-[1-14C]glucose. Insulin-stimulated glucose uptake was normal in hearts expressing 15% of normal GLUT4 levels but markedly reduced in mice with more profound reduction in GLUT4. Cardiac enlargement occurred only when GLUT4 levels were less than 5% of normal values. In heart there is a threshold level of GLUT4 above which insulin-stimulated glucose uptake is maintained. As little as 5% of normal GLUT4 levels expressed in heart is sufficient to prevent the development of cardiac hypertrophy.

Exploring barriers to uptake of breast cancer screening in South East London : a study of women's views.

Research on both sides of the Atlantic demonstrates that achieving high uptake of breast cancer screening remains an important area of public heath concern. UK government targets for breast screening uptake are 70%, however, much lower figures are found in many parts of the country, including South East London. This paper reports the findings of a study carried out to explore the views of women aged 50 to 64 (the age group covered by the free National Health Service screening programme) in order to: · establish in what way women who do not attend for screening are different from women who do attend · ascertain the views of the non-attenders with a view to making recommendations to the service which may help address the low uptake locally.

305 women were recruited through a variety of different community sources across the study area. Using a structured questionnaire/interview, women gave their views on their health concerns generally, as well as on breast screening in particular. The analysis (being undertaken now, to be completed by May 2005) will explore the influence of candidacy (women's assessment of the personal risk to them of their disease) on women's screening behaviour and the differences, if any, between the major ethnic groups in the area, indigenous white, black African and black Caribbean.

Learning Objectives:
# At the conclusion of the session, participants will be able to

* 1. Describe the factors associated with women’s screening behaviour
* 2. Evaluate the relevance of candidacy in understanding screening behaviour
* 3. Assess the relevance of UK findings for screening programmes elsewhere.