Relationship of nutritional and metabolic factors to non-invasive indices of macrovascular disease in diabetes

Factors which may account for the high frequency of macrovascular disease in diabetics are age, sex, cigarette smoking, hypertension, obesity, lack of exercise, diet, hyperglycaemia, hyperinsulinaeroia, hypercholesterolaemia, hypertriglyceridaemia, low HDL-cholesterol concentration, elevated free fatty acid concentration and enhanced platelet aggregation. Twenty seven (13 men and 14 women) non-insulin-dependent diabetics and thirty eight age, height and weight matched healthy subjects (10 men and 28 women) were studied. None of the subjects were smokers, or hypertensive. No subject had any clinical evidence of peripheral arterial disease, coronary heart disease or cerebrovascular disease. All had apparently normal peripheral pulses and normal ankle/arm blood pressure indices. Methods for determining arterial compliance in the segment between the left subclavian artery and each common femoral artery, and proximal resistance at the common femoral artery and posterior tibial artery, have been reviewed and developed. An appropriate food intake methodology for deriving food indices from food records was developed. Biochemical determinants have been made of glucose tolerance, glycosylated haemoglobin, serum total cholesterol, HDL-cholesterol, LDL-cholesterol, triglyceride, plasma free fatty acid and insulin. A significant decrease in the arterial compliance, and a significant increase in the arterial proximal resistance at the common femoral artery and posterior tibial artery in non-insulin-dependent diabetics, compared with their healthy controls, have been found. Significant negative correlation between arterial compliance and proximal resistance and, a significant positive correlation between the arterial proximal resistance of common femoral artery and posterior tibial artery were found. Differences between control (healthy subjects) and non-insulin-dependent diabetic groups indicate that preclinical peripheral arterial disease can be recognised even in mild diabetics by non-invasive measurement of arterial compliance or proximal resistance. There were significant and negative correlations between arterial compliance and each of blood glucose, blood glycosylated haemoglobin (HbAlC), plasma free fatty acid and plasma insulin concentration. There were significant and positive correlations between arterial proximal resistance of common femoral artery and posterior tibial artery and each of blood glucose, glycosylated haemoglobin and plasma free fatty acid concentration. Multivariate analysis to examine each of the biochemical factors Including blood glucose, blood glycosylated haemoglobin (HbAlC), plasma free fatty acid, plasma Insulin and lipids, showed that the factor which most influenced the arterial compliance and the proximal resistance of posterior tibial artery was the glucose level in the fasting state or the glucose response after a glucose load. In addition, the factors which most influenced proximal resistance of the common femoral artery were free fatty acid -level in the fasting state or glucose response after a glucose load. The factors which most influenced arterial compliance were glucose level in men, and the insulin level in the fasting state or the plasma free fatty acid response after a glucose load in women. These findings indicate that blood glucose, plasma free fatty acid and plasma insulin are risk factors for changes in arterial wall characteristic at a stage when no clinical evidence of macrovascular disease is apparent. Arterial compliance was decreased and the proximal resistance of posterior tibial artery was increased in those with a low intake of protective foods compared with those with a high intake whether healthy subjects or non-insulin-dependent diabetics. Arterial compliance was decreased in non-fish eaters compared with the fish eaters whether healthy subjects or non-insulin-dependent diabetics. Proximal resistance of the posterior tibia! artery in non-fish eaters was increased compared with fish eaters in healthy subjects. Overall, food variety, a protective food score consumption and fish consumption emerge as importance determinants of arterial wall characteristics at a stage when no clinical evidence of macrovascular disease is apparent.

Single-point isotope measurements in blood cells and plasma to estimate the time since diet switches

1. Understanding ecological phenomena often requires an accurate assessment of the timing of events. To estimate the time since a diet shift in animals without knowledge on the isotope ratios of either the old or the new diet, isotope ratio measurements in two different tissues (e.g. blood plasma and blood cells) at a single point in time can be used. For this ‘isotopic-clock’ principle, we present here a mathematical model that yields an analytical and easily calculated outcome.

2. Compared with a previously published model, our model assumes the isotopic difference between the old and new diets to be constant if multiple measurements are taken on the same subject at different points in time. Furthermore, to estimate the time since diet switch, no knowledge of the isotopic signature of tissues under the old diet, but only under the new diet is required.

3. The two models are compared using three calibration data sets including a novel one based on a diet shift experiment in a shorebird (red knot Calidris canutus); sensitivity analyses were conducted. The two models behaved differently and each may prove rather unsatisfactory depending on the system under investigation. A single-tissue model, requiring knowledge of both the old and new diets, generally behaved quite reliably.

4. As blood (cells) and plasma are particularly useful tissues for isotopic-clock research, we trawled the literature on turnover rates in whole blood, cells and plasma. Unfortunately, turnover rate predictions using allometric relations are too unreliable to be used directly in isotopic-clock calculations.

5. We advocate that before applying the isotopic-clock methodology, the propagation of error in the ‘time-since-diet-shift’ estimation is carefully assessed for the system under scrutiny using a sensitivity analysis as proposed here.

Ultrastructural analysis of plastids in angiosperm pollen tubes

When Rhododendron pollen tubes are cultured in the dark, electron-dense bodies are present that appear to be a metabolically altered form of a proplastid that is difficult to fix for electron microscopy, and whose membranes may not be intact. When similar pollen tubes are cultured in a dark/light regime, ultrastructurally well-defined proplastids are present after fixation in glutaraldehyde with PIPES buffer and tannic acid, followed by osmic acid. This fixation technique also gave the best ultrastructural images of those proplastids in pollen tubes grown in the dark. Pollen tube plastids have the potential to become chromoplasts when cultured in a dark/light regime as evidenced by the presence of branched tubules characteristic of these organelles. Light appears to be a hitherto neglected environmental factor involved in regulating pollen tube growth. This improved fixation procedure demonstrates the bilayered nature of the membranes surrounding sperm cells and the existence of cytoplasmic channels connecting sperm cell and pollen tube plasma membranes.

Functional analysis of proteins involved in Plasmodium falciparum merozoite invasion of red blood cells

Plasmodium falciparum causes the most lethal form of malaria in humans and is responsible for over two million deaths per year. The development of a vaccine against this parasite is an urgent priority and potential protein targets include those on the surface of the asexual merozoite stage, the form that invades the host erythrocyte. The development of methods to transfect P. falciparum has enabled the construction of gain-of-function and loss-of-function mutants and provided new strategies to analyse the role of parasite proteins. In this review, we describe the use of this technology to examine the role of merozoite antigens in erythrocyte invasion and to address their potential as vaccine candidates.

Ageing effect of plasma-treated wool

Atmospheric pressure plasma treatment of wool fabric, with a relatively short exposure time, effectively removed the covalently bonded lipid layer from the wool surface. The plasma-treated fabric showed increased wettability and the fibres showed greater roughness. X-ray photoelectron spectroscopy (XPS) analysis showed a much more hydrophilic surface with significant increases in oxygen and nitrogen concentrations and a decrease in carbon concentration. Adhesion, as measured by scanning probe microscopy (SPM) force volume analysis, also increased, consistent with the more hydrophilic surface leading to a greater meniscus force on the SPM probe. The ageing of fibres from the plasma-treated fabric was assessed over a period of 28 days. While no physical changes were observed, the chemical nature of the surface changed significantly. XPS showed a decrease in the hydrophilic nature of the surface with time, consistent with the measured decrease in wettability. This change is proposed to be due to the reorientation of proteolipid chains. SPM adhesion studies also showed the surface to be changing with time. After ageing for 28 days, the plasma-treated surface was relatively stable and still dramatically different from the untreated fibre, suggesting that the oxidation of the surface and modification or removal of the lipid layer were permanent.

Influence of atmospheric helium plasma on the surface energy of jute fibres and the performance of resulting composites

In this work, an atmospheric pressure glow discharge helium plasma treatment was employed to modify the surface properties of jute fibres. The resulting bio-composites showed an increase in flexural properties and interlaminar shear strength (ILSS) compared to composites produced using untreated jute fibres. To understand the reason behind the ILSS improvement, the acid–base properties of jute fibres were determined by contact angle analysis using the capillary rise method. The results were fitted further to van Oss–Chaudhury–Good (vOCG) and Chang–Qin–Chen (CQC) models to determine the Lifshitz–van der Waals (LW) and acid–base components of surface energy. Surface energy determined by the vOCG model revealed that plasma treatment of jute fibre resulted in a 22% increase in total surface energy, a 19% increase in the LW component and a 24% increase in the acid–base component of surface energy. The increase in the acid–base component is due to the significant increase (69%) in the electron-accepting (γ+S) parameter. On the other hand, the CQC model clearly indicates an amphoteric nature of the fibre surface based on opposite signs of the acid and base principal values (PSa and PSb). Overall, the results indicated that increases in both LW and acid–base components were responsible for improvement in the properties of the composites.

Deposition and wetting characteristics of polyelectrolyte multilayers on plasma-modified porous polyethylene

Hydrophilic and chemically reactive porous media were prepared by adsorbing functional polymers at the surface of sintered polyethylene membranes. Modification of the membrane was accomplished by first exposing the membrane to an oxygen glow discharge gas plasma to introduce an electrostatic charge at the membrane surfaces. Cationic polyelectrolyte polyethylenimine (PEI) was adsorbed from solution to the anionic-charged surface to form an adsorbed monolayer. The adsorption of a second anionic polyelectrolyte onto the PEI layer allows further modification of the membrane surface to form a polyelectrolyte-bilayer complex. The conformation and stability of the adsorbed monolayers and bilayers comprising the modified surface are probed as a function of the polymer structure, charge density, and solubility. Using X-ray photoelectron spectroscopy analysis, we demonstrate that the presence of the polyelectrolyte multilayers drastically increases the density and specificity of the functional groups at the surface, more than what can be achieved through the plasma modification alone. Also, using the wicking rate of deionized, distilled water through the porous membrane to gauge the interfacial energy of the modified surface, we show that the membrane wicking rate can be controlled by varying the chemistry of the adsorbing polyelectrolytes and, to a lesser extent, by adjusting the polarity or ionic strength of the polyelectrolyte solution.

Rapid development of non-alcoholic steatohepatitis in Psammomys obesus (Israeli Sand Rat)

Background and AimsA major impediment to establishing new treatments for non-alcoholic steatohepatitis is the lack of suitable animal models that accurately mimic the biochemical and metabolic characteristics of the disease. The aim of this study was to explore a unique polygenic animal model of metabolic disease as a model of non-alcoholic steatohepatitis by determining the effects of 2% dietary cholesterol supplementation on metabolic and liver endpoints in Psammomys obesus (Israeli sand rat).MethodsP. obesus were provided ad libitum access to either a standard rodent diet (20% kcal/fat) or a standard rodent diet supplemented with 2% cholesterol (w/w) for 4 weeks. Histological sections of liver from animals on both diets were examined for key features of non-alcoholic steatohepatitis. The expression levels of key genes involved in hepatic lipid metabolism were measured by real-time PCR.ResultsP. obesus fed a cholesterol-supplemented diet exhibited profound hepatomegaly and steatosis, and higher plasma transaminase levels. Histological analysis identified extensive steatosis, inflammation, hepatocyte injury and fibrosis. Hepatic gene expression profiling revealed decreased expression of genes involved in delivery and uptake of lipids, and fatty acid and triglyceride synthesis, and increased expression of genes involved in very low density lipoprotein cholesterol synthesis, triglyceride and cholesterol export.ConclusionsP. obesus rapidly develop non-alcoholic steatohepatitis when fed a cholesterol-supplemented diet that appears to be histologically and mechanistically similar to patients.

Peripheral brain-derived neurotrophic factor in schizophrenia and the role of antipsychotics: meta-analysis and implications

It has been postulated that schizophrenia (SZ) is related to a lower expression of brain-derived neurotrophic factor (BDNF). In the past few years, an increasing number of divergent clinical studies assessing BDNF in serum and plasma have been published. It is now possible to verify the relationship between BDNF levels and severity of symptoms in SZ as well as the effects of antipsychotic drugs on BDNF using meta-analysis. The aims of this study were to verify if peripheral BDNF is decreased in SZ, whether its levels are correlated with positive and negative symptomatology and if BDNF levels change after antipsychotic treatment. This report consists of two distinct meta-analyses of peripheral BDNF in SZ including a total of 41 studies and more than 7000 participants: (1) peripheral BDNF levels in serum and plasma were moderately reduced in SZ compared with controls. Notably, this decrease was accentuated with the disease duration. However, the extent of peripheral BDNF level decrease did not correlate with the severity of positive and negative symptoms. (2) In plasma, but not serum, peripheral BDNF levels are consistently increased after antipsychotic treatment irrespective of the patient's response to medication. In conclusion, there is compelling evidence that there are decreased levels of peripheral BDNF in SZ, in parallel to previously described reduced cerebral BDNF expression. It remains unclear whether these systemic changes are causally related to the development of SZ or if they are merely a pathologic epiphenomenon.Molecular Psychiatry advance online publication, 30 September 2014; doi:10.1038/mp.2014.117.

In vitro and in situ screening systems for morphological and phytochemical analysis of Withania somnifera germplasms

We report, for the first time for Withania somnifera, the use of a modified in vitro system for morphological and phytochemical screening of true to type plants as compared with those grown in a conventional in situ system. Eleven germplasms of cultivated W. somnifera from different regions of India were collected to examine chemotypic variation in withaferin A (WA). Methods were developed to optimize WA extraction. The maximum concentration of WA was extracted from manually ground leaf and root material to which 60 % methanol was added followed by sonication in a water bath sonicator. Variation in WA concentration in whole plants was observed amongst the different germplasms. In the in vitro system, the concentration of WA ranged between 0.27 and 7.64 mg/g dry weight (DW) and in the in situ system, the range in concentration was between 8.06 and 36.31 mg/g DW. The highest amount of WA found in leaves was 7.37 and 41.42 mg/g DW in the in vitro and the in situ systems respectively. In roots, the highest WA concentration was 0.27 mg/g DW in the in vitro and 0.60 mg/g DW in the in situ system. There are distinct advantages in using the in vitro grown plants rather than those grown in the in situ system including the simplicity of design, efficient use of space and nutrition and a system which is soil and contaminant free. The proposed in vitro system is therefore ideal for utilization in molecular, enzymatic and biochemical studies.

(1)H-NMR analysis of the human urinary metabolome in response to an 18-month multi-component exercise program and calcium-vitamin-D3 supplementation in older men

The musculoskeletal benefits of calcium and vitamin-D3 supplementation and exercise have been extensively studied, but the effect on metabolism remains contentious. Urine samples were analyzed by (1)H-NMR spectroscopy from participants recruited for an 18-month, randomized controlled trial of a multi-component exercise program and calcium and vitamin-D3 fortified milk consumption. It was shown previously that no increase in musculoskeletal composition was observed for participants assigned to the calcium and vitamin-D3 intervention, but exercise resulted in increased bone mineral density, total lean body mass, and muscle strength. Retrospective metabolomics analysis of urine samples from patients involved in this study revealed no distinct changes in the urinary metabolome in response to the calcium and vitamin-D3 intervention, but significant changes followed the exercise intervention, notably a reduction in creatinine and an increase in choline, guanidinoacetate, and hypoxanthine (p < 0.001, fold change > 1.5). These metabolites are intrinsically involved in anaerobic ATP synthesis, intracellular buffering, and methyl-balance regulation. The exercise intervention had a marked effect on the urine metabolome and markers of muscle turnover but none of these metabolites were obvious markers of bone turnover. Measurement of specific urinary exercise biomarkers may provide a basis for monitoring performance and metabolic response to exercise regimes.

Sliding wear behavior of plasma sprayed Fe3Al–Al2O3 graded coatings

Fe3Al–Al2O3 double-layer coatings (DC), Fe3Al-Fe3Al/50%Al2O3–Al2O3 triple-layer coatings (TC) and Fe3Al-Al2O3 graded coatings (GC) were produced from a series of Fe3Al/Al2O3 composite powders with different compositions on low carbon steel substrate using PLAXAIR plasma spraying equipment. Friction behaviors and wear resistance of the three kinds of coatings have been investigated under different loads. Tests were carried out using an MRH-3 standard machine, in lineal contact sliding under dry condition against hardmetal, at a sliding velocity of about 1.57 ms−1. Wear rates under different loads were measured and the friction coefficients were recorded. SEM analysis was carried out to identify the wear mechanisms. The results show that the GC has higher wear-resistance than DC and TC. The tribological characteristics of graded coating were different along the depth of the coatings, and the surface of coatings with pure Al2O3 does not show the best wear resistance. The wear rate and friction coefficients were also different under different loads. The failure types of plasma-sprayed Fe3Al-Al2O3 graded coatings in lineal contact were: loosening of ceramic particles, crack nucleation and propagation, brittle fracture, plastic deformation, and adhesive wear.

Short term fat feeding rapidly increases plasma insulin but does not result in dyslipidaemia

Although the association between obesity and hypertension is well known, the underlying mechanism remains elusive. Previously, we have shown that 3 week fat feeding in rabbits produces greater visceral adiposity, hypertension, tachycardia and elevated renal sympathetic nerve activity compared to rabbits on a normal diet. Because hyperinsulinaemia, hyperleptinemia and dyslipidaemia are independent cardiovascular risk factors associated with hypertension we compared plasma insulin, leptin and lipid profiles in male New Zealand White rabbits fed a normal fat diet (NFD 4.3% fat, n = 11) or high fat diet (HFD 13.4% fat, n = 13) at days 1, 2, 3 and weeks 1, 2, 3 of the diet. Plasma concentrations of diacylglyceride (DG), triacylglyceride (TG), ceramide and cholesteryl esters (CE) were obtained after analysis by liquid chromatography mass spectrometry. Plasma insulin and glucose increased within the first 3 days of the diet in HFD rabbits (P <0.05) and remained elevated at week 1 (P <0.05). Blood pressure and heart rate followed a similar pattern. By contrast, in both groups, plasma leptin levels remained unchanged during the first few days (P >0.05), increasing by week 3 in fat fed animals alone (P <0.05). Concentrations of total DG, TG, CE and Ceramide at week 3 did not differ between groups (P >0.05). Our data show plasma insulin increases rapidly following consumption of a HFD and suggests that it may play a role in the rapid rise of blood pressure. Dyslipidaemia does not appear to contribute to the hypertension in this animal model.