Development of impedimetric and optical calcium biosensor by using modified gold electrode with porcine S100A12 protein

We describe the development of a label free method to analyze the interactions between Ca(2+) and the porcine S100A12 protein immobilized on polyvinyl butyral (PVB). The modified gold electrodes were characterized using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM) and surface plasmon resonance (SPR) techniques. SEM analyses of PVB and PVB-S100A12 showed a heterogeneous distribution of PVB spherules on gold surface. EIS and CV measurements have shown that redox probe reactions on the modified gold electrodes were partially blocked due the adsorption of PVB-S100A12, and confirm the existence of a positive response of the immobilized S100Al2 to the presence of calcium ions. The biosensor exhibited a wide linear response to Ca(2+) concentrations ranging from 12.5 to 200 mM. The PVB-S100A12 seems to be bound to the gold electrode surface by physical adsorption: we observed an increase of 1184.32 m degrees in the SPR angle after the adsorption of the protein on the PVB surface (in an indication that 9.84 ng of S100A12 are adsorbed per mm(2) of the Au-PVB electrode), followed by a further increase of 581.66 m degrees after attachment of the Ca(2+) ions. In addition, no SPR response is obtained for non-specific ions. These studies might be useful as a platform for the design of new reusable and sensitive biosensing devices that could find use in the clinical applications. (C) 2010 Elsevier B.V. All rights reserved.

Study on the observation of Eu(2+) and Eu(3+) valence states in low silica calcium aluminosilicate glasses

The optical, magnetic and structural properties of Eu doped low silica calcium aluminosilicate glasses were investigated. The optical absorption coefficient presented two bands at 39 246 and 29 416 cm(-1), which were assigned respectively to the 4f(7) ((8)S(7/2)) -> 4f(6) (4F(J)) 5d (T(2g)), and 4f(7) ((8)S(7/2)) -> 4f(6) (4F(J)) 5d (E(g)) transitions of Eu(2+). The fluorescence measured at 300 K on a sample doped with 0.5 wt% of Eu(2)O(3) exhibited a broad band centered at 17 350 cm(-1), which is attributed to the 4f(6)5d -> 4f(7) transition of Eu(2+), whereas the additional peaks are due to the (5)D(0) -> (7)F(J) (J = 1, 2, 4) transitions of Eu(3+). From magnetization and XANES data it was possible to evaluate the fractions of Eu(2+) and Eu(3+) for the sample doped with 0.5 and 5.0 wt% of Eu(2)O(3), the values of which were approximately 30 and 70%, respectively.

Structure and Calcium-Binding Activity of LipL32, the Major Surface Antigen of Pathogenic Leptospira sp.

Leptospixosis, a spirochaetal zoonotic disease caused by Leptospira, has been recognized as an important emerging infectious disease. LipL32 is the major exposed outer membrane protein found exclusively in pathogenic leptospires, where it accounts for up to 75% of the total outer membrane proteins. It is highly immunogenic, and recent studies have implicated LipL32 as an extracellular matrix binding protein, interacting with collagens, fibronectin, and laminin. In order to better understand the biological role and the structural requirements for the function of this important lipoprotein, we have determined the 2.25-angstrom-resolution structure of recombinant LipL32 protein corresponding to residues 21-272 of the wild-type protein (LipL32(21-272)). The LipL32(21-272) monomer is made of a jelly-roll fold core from which several peripheral secondary structures protrude. LipL32(21-272) is structurally similar to several other jelly-roll proteins, some of which bind calcium ions and extracellular matrix proteins. Indeed, spectroscopic data (circular dichroism, intrinsic tryptophan fluorescence, and extrinsic 1-amino-2-naphthol-4-sulfonic acid fluorescence) confirmed the calcium-binding properties of LipL32(21-272). Ca(2+) binding resulted in a significant increase in the thermal stability of the protein, and binding was specific for Ca(2+) as no structural or stability perturbations were observed for Mg(2+), Zn(2+), or Cu(2+). Careful examination of the crystal lographic structure suggests the locations of putative regions that could mediate Ca(2+) binding as well as binding to other interacting host proteins, such as collagens, fibronectin, and lamixidn. (C) 2009 Elsevier Ltd. All rights reserved.

The effect of calcium salts on the viscosity and adsorption behavior of xanthan

The effect of CaCl(2), Ca(NO(3))(2), CaSO(4), CaCO(3) and Ca(3)(PO(4))(2) on the flow behavior of xanthan gum solutions was investigated. Regardless the concentration and type of calcium salt used, xanthan solutions presented pseudoplastic behavior. The soluble salts (CaCl(2) and Ca(NO(3))(2)) induced the disordered state in the xanthan chains at concentration of 1.0 g/L or 10 g/L, decreasing the flow consistency index (K) values. At 100 g/L soluble salts K values were similar to those found for pure xanthan solutions, whereas at the same concentration of insoluble particles the K values increased 20%. The adsorption of xanthan gum onto Si/SiO(2) surfaces in the presence of calcium salts was investigated by ellipsometry and atomic force microscopy (AFM). The adsorbed layer of xanthan onto Si/SiO(2) consisted of two regions: (i) a thin acid resistant sublayer, where xanthan chains were like highly entangled fibers and (ii) a thick upperlayer, whose morphology was calcium salt dependent. (C) 2010 Elsevier Ltd. All rights reserved.

Effect of Calcium Pre-rinse and Fluoride Dentifrice on Enamel and on Dental Plaque Formed In Situ

Purpose: The aim of this in situ double-blind randomised crossover study was to investigate the effect of calcium (Ca) pre-rinse on the composition of plaque and on enamel prior to the use of fluoride (F) dentifrice. Materials and Methods: During four phases (14 days each) of this study, 10 volunteers had agreed to wear dental appliances containing two healthy bovine enamel blocks. A fresh solution containing 20% weight/volume (w/v) sucrose was dripped on the enamel blocks ex vivo for 5 min three times a day. Subsequently, the appliances were replaced in the mouth, and the volunteers rinsed their mouth with 10 mL of a Ca (150 mmol/L) or a placebo rinse (1 min). In sequence, a slurry (1:3 w/v) of F (1030 ppm) or placebo dentifrice was dripped onto the blocks ex vivo for 1 min. During this time, the volunteers brushed their teeth with the respective dentifrice. The appliances were replaced in the mouth, and the volunteers rinsed their mouth with water. The plaque formed on the blocks was analysed for F and Ca. The enamel demineralisation as well as the incorporation of F on enamel was evaluated by cross-sectional microhardness and alkali-soluble F analysis, respectively. Data were tested using analysis of variance (P < 0.05). Results: The Ca pre-rinse prior to the use of the F dentifrice led to a three- and sixfold increase in the plaque F and Ca concentrations, respectively. It also did not have any additive effect on the F content on the enamel and the demineralisation of the enamel, in comparison with the use of F dentifrice alone. Conclusions: A Ca lactate rinse used prior to the F dentifrice was able to change the mineral content in the plaque, but it was unable to prevent enamel demineralisation.

Knowledge about folate and the contribution of nutrient supplements to the intake of folic acid and vitamin B12 in Australian adults in 1995 and 1996

The present paper reports knowledge about folate and the contribution of supplements to the intake of folic acid and vitamin B12 in Australian adults during 1995 and 1996. Data were obtained from two population survey monitor surveys conducted in a sample of 5422 adults in 1995-96. The surveys were designed to complement food intake data from the 1995 National Nutrition Survey and to provide an estimate of total folate intake prior to the implementation of voluntary fortification of foods with folic acid. The proportion with knowledge about folate increased with education level and socioeconomic status and was greater in women than men. It was also greater in those who were married, and in those residing in Western Australia and the Australian Capital Territory than in other states and territories. Five per cent of men and 10% of women had taken a supplement containing folio acid on the previous day. The equivalent figures for vitamin B12 were 7.5% for men and 12.5% for women. On average, intake of folic acid from supplements was 11 micro g per day for men and 28 micro g per day for women and intake of vitamin B12 was 2.6 micro g per day for men and 4.5 micro g per day for women. For individuals who consumed a supplement containing folic acid on the day before the survey the median folic acid contribution was 200 micro g. In 1995 and 1996 only one in two adult Australians had heard of folate and only one in ten women of child-bearing age had taken a supplement containing folic acid.

A practical approach to monitoring nutrient supplement intake of Australian adults

The adoption, in mid-1995, of the revised food Standard A9, which permits the more liberal addition of nutrients to a range of food products, highlighted the need to obtain information on nutrient intake from supplements to complement the i 995 National Nutrition Survey data on nutrient intake from food. This paper describes the method used to obtain quantitative information on nutrient supplement intake and reports on the prevalence of supplement use in different subgroups of the Australian population. Information on supplement intake was obtained in two Australian Bureau of Statistics Population Survey Monitor surveys in August 1995 and February 1996 using the Therapeutic Goods Administration (TGA) registration numbers to identify individual products. Approximately 18% of men and 29% of women aged 18 years and over reported consuming a nutrient supplement on the day before the survey and these proportions increased to 25% and 35% respectively for consumption during the two weeks before the survey. The prevalence of supplement intake increased with age, education level, socioeconomic status, employment status and with fruit and vegetable intake. The substantial proportion of Australian adults who consume nutrient supplements, and the rapidly changing composition of the Australian food supply in response to changes in food regulation, indicate that there is a need for regular monitoring of nutrient intake from supplements. The use of TGA registration numbers to identify supplements provides a practical way to address this need.

Vitamin D intake and vitamin D status of Australians

The main source of vitamin D for Australians is exposure to sunlight. Thus, levels of serum 25-hydroxyvitamin D3, the indicator of vitamin D status, vary according to the season and are lower at the end of winter.

In Australia and New Zealand, the prevalence of vitamin D deficiency varies, but is acknowledged to be much higher than previously thought. One study found marginal deficiency in 23% of women, and another frank deficiency in 80% of dark-skinned and veiled women. The groups at greatest risk of vitamin D deficiency in Australia are dark-skinned and veiled women (particularly in pregnancy), their infants, and older persons living in residential care.

Only a few foods (eg, fish with a high fat content) contain significant amounts of vitamin D. In Australia, margarine and some milk and milk products are currently fortified with vitamin D.

The average estimated dietary intake of vitamin D for men is 2.6–3.0 µg/day and for women is 2.0–2.2 µg/day. The estimated dietary requirement of vitamin D is at least 5.0 µg/day and may be higher for older people.

Adequate intake of vitamin D is unlikely to be achieved through dietary means, particularly in the groups at greatest risk, although vitamin D-fortified foods may assist in maintaining vitamin D status in the general population.

An appropriate health message for vitamin D needs to balance the need for sunshine against the risk of skin cancer.

Calcium chemistry of Blue Lake, Mt Gambier, Australia, and relevance to remarkable seasonal colour changes

Calcium speciation and other water quality variables in the Blue Lake, Mt Gambier, Australia, were monitored between August 1999 and August 2000 in order to test previously proposed mechanisms for the seasonal colour changes of this lake. The concentration of calcite was found to be highest in winter when the lake appears grey, and lowest in summer when the lake appears blue. A potential component of the colour change mechanism is therefore identified in which the lake is grey in winter because of non-selective scattering of light by calcite particles, and blue in summer because of the absence of absorbing or scattering impurities.

Nicotine treatment decreases food intake and body weight via a leptin-independent pathway in Psammomys obesus

It has been reported previously that leptin may be involved in nicotine's ability to reduce body weight. Our aim was to investigate whether the anorexic action of nicotine is related to the actions of leptin by utilizing lean leptin-sensitive and obese leptin-resistant Psammomys obesus. Lean and obese P. obesus were assigned to receive nicotine sulphate at 6, 9 or 12 mg/day or saline (control) for 9 days (n = 6-10 in each group), administered using mini-osmotic pumps. Food intake, body weight, plasma leptin concentrations, plasma insulin and blood glucose were measured at baseline and throughout the study period. Nicotine treatment reduced food intake by up to 40% in lean and obese P. obesus. Plasma leptin levels fell significantly only in lean nicotine-treated animals, whereas no changes were observed in obese nicotine-treated animals. However, both lean and obese nicotine-treated animals had similar reductions in body weight. Our results show that nicotine has dramatic effects on food intake and body weight, however, these changes appear to be independent of the leptin signalling pathway.

Foods prepared outside the home: association with selected nutrients and body mass index in adult Australians

Objective: To determine the proportion of energy from foods prepared outside the home (FPOH) and the relationships with energy and nutrient intakes and body mass index (BMI).

Design: A nutrition survey of a representative sample of the Australian population aged 18 years and over (n = 10 863). Measure used was a 24-hour dietary recall. Underreporters (energy intake/estimated basal metabolic rate (EI/BMR) <0.9) were excluded from analysis. Daily energy and selected nutrient intakes were calculated using a 1996 nutrient composition database for all foods/beverages during the 24-hour period.

Results: On average FPOH contributed a significant 13% to total energy intake. About a third of the sample had consumed FPOH in the last 24 hours and on average this group consumed a third of their total energy as FPOH. The relative contributions of fat (for men and women) and alcohol (for women) were significantly higher for those in the top tertile of FPOH consumers. The intakes of fibre and selected micronutrients (calcium, iron, zinc, folate and vitamin C) were significantly lower in this group. After adjustment for age and income no relationship between FPOH and BMI was observed.

Conclusions: FPOH make a significant contribution to the energy intake of a third of the Australian population. FPOH contribute to poor nutritional intakes. Altering the supply of FPOH may be an effective means of improving diets at a population level.

Effect of alcohol intake on muscle glycogen storage after prolonged exercise

We studied the effects of alcohol intake on postexercise muscle glycogen restoration with samples from vastus lateralis being collected immediately after glycogen-depleting cycling and after a set recovery period. Six well-trained cyclists undertook a study of 8-h recovery (2 meals), and another nine cyclists undertook a separate 24-h protocol (4 meals). In each study, subjects completed three trials in crossover order: control (C) diet [meals providing carbohydrate (CHO) of 1.75 g/kg]; alcohol-displacement (A) diet (1.5 g/kg alcohol displacing CHO energy from C) and alcohol + CHO (AC) diet (C + 1.5 g/kg alcohol). Alcohol intake reduced postmeal glycemia especially in A trial and 24-h study, although insulin responses were maintained. Alcohol intake increased serum triglycerides, particularly in the 24-h study and AC trial. Glycogen storage was decreased in A diets compared with C at 8 h (24.4 ± 7 vs. 44.6 ± 6 mmol/kg wet wt, means ± SE, P < 0.05) and 24 h (68 ± 5 vs. 82 ± 5 mmol/kg wet wt, P < 0.05). There was a trend to reduced glycogen storage with AC in 8 h (36.2 ± 8 mmol/kg wet wt, P = 0.1) but no difference in 24 h (85 ± 9 mmol/kg wet wt). We conclude that 1) the direct effect of alcohol on postexercise glycogen synthesis is unclear, and 2) the main effect of alcohol intake is indirect, by displacing CHO intake from optimal recovery nutrition practices.

Energy density of foods and beverages in the Australian food supply: influence of macronutrients and comparison to dietary intake

Objectives: The energy density (ED) of the diet is considered an important determinant of total energy intake and thus energy balance and weight change. We aimed to compare relationships between ED and macronutrient content in individual food and beverage items as well as population diet in a typical Western country. Design: Nutrient data for 3673 food items and 247 beverage items came from the Australian Food and Nutrient database (AusNut). Food and beverage intake data came from the 1995 Australian National Nutrition Survey (a 24-h dietary recall survey in 13 858 people over the age of 2). Relationships between ED and macronutrient and water content were analysed by linear regression with 95% prediction bands. Results: For both individual food items and population food intake, there was a positive relationship between ED and percent energy as fat and negative relationships between ED and percent energy as carbohydrate and percent water by weight. In all cases, there was close agreement between the slopes of the regression lines between food items and dietary intake. There were no clear relationships between ED and macronutrient content for beverage items. Carbohydrate (mostly sucrose) contributed 91, 47, and 25% of total energy for sugar-based, fat-based, and alcohol-based beverages respectively. Conclusions: The relationship between ED and fat content of foods holds true across both population diets and individual food items available in the food supply in a typical Western country such as Australia. As high-fat diets are associated with a high BMI, population measures with an overall aim of reducing the ED of diets may be effective in mediating the growing problem of overweight and obesity.

Effect of fat replacement by insulin or lupin-kernal fibre on sausage patty acceptability, post-meal perceptions of satiety and food intake in men

The present study examined whether replacing fat with inulin or lupin-kernel fibre influenced palatability, perceptions of satiety, and food intake in thirty-three healthy men (mean age 52 years, BMI 27·4 kg/m²), using a within-subject design. On separate occasions, after fasting overnight, the participants consumed a breakfast consisting primarily of either a full-fat sausage patty (FFP) or a reduced-fat patty containing inulin (INP) or lupin-kernel fibre (LKP). Breakfast variants were alike in mass, protein and carbohydrate content; however the INP and LKP breakfasts were 36 and 37 % lower in fat and 15 and 17 % lower in energy density respectively compared with the FFP breakfast. The participants rated their satiety before breakfast then evaluated patty acceptability. Satiety was rated immediately after consuming the breakfast, then over the subsequent 4·5 h whilst fasting. Food consumed until the end of the following day was recorded. All patties were rated above ‘neither acceptable or unacceptable’, however the INP rated lower for general acceptability (P=0·039) and the LKP lower for flavour (P=0·023) than the FFP. The LKP breakfast rated more satiating than the INP (P=0·010) and FFP (P=0·016) breakfasts. Total fat intake was 18 g lower on the day of the INP (P=0·035) and 26 g lower on the day of the LKP breakfast (P=0·013) than the FFP breakfast day. Energy intake was lower (1521 kJ) only on the day of the INP breakfast (P=0·039). Both inulin and lupin-kernel fibre appear to have potential as fat replacers in meat products and for reducing fat and energy intake in men.