Skeletal muscle total creatine content and creatine transporter gene expression in vegetarians prior to and following creatine supplementation

This study examined the effect of vegetarianism on skeletal muscle total creatine (TCr) content and creatine transporter (CreaT) gene expression, prior to and during 5 d of Cr supplementation (CrS). In a double-blind, crossover design, 7 vegetarians (VEG) and nonvegetarians (NVEG) were assigned Cr or placebo supplements for 5 d and after 5 wk, received the alternative treatment. Muscle sampling occurred before, and after 1 and 5 d of treatment ingestion. Basal muscle TCr content was lower (P < 0.05) in VEG compared with NVEG. Muscle TCr increased (P < 0.05) throughout the Cr trial in both groups but was greater (P < 0.05) in VEG compared with NVEG, at days 1 and 5. CreaT gene expression was not different between VEG and NVEG. The results indicate that VEG have a lower muscle TCr content and an increased capacity to load Cr into muscle following CrS. Muscle CreaT gene expression does not appear to be affected by vegetarianism.

A simple 96-well microplate method for estimation of total polyphenolic content in seaweeds

Seaweed polyphenols are potent antioxidants and have also been shown to have α-glucosidase inhibiting activity. In our continuous efforts to develop new marine-based nutraceuticals and functional food ingredients, we have investigated many algal species collected on the Atlantic coast of Canada. A simple method for estimating the total polyphenol content in seaweeds and their extracts was developed based on the classic Folin-Ciocalteau colorimetric reaction. By using the 96-well microplate and a microplate reader, this new method saves experimental time, significantly reduces the amount of sample required, handles large number of samples in one experiment, and also improves the repeatability of the results.

A number of algal samples collected on the seashore of Nova Scotia, Canada, were analyzed for their levels of polyphenol content using this microplate-based method. The antioxidant activity of these samples was also assessed by using DPPH (2, 2-diphenyl-1-picrylhydrazyl) radical scavenging assay. The results showed that there is a strong correlation between the total polyphenol content and the potency of antioxidant effect.

Ethephon increases rubber tree latex yield by regulating aquaporins and alleviating the tapping-induced local increase in latex total solid content

The concentration of phloem solute generally falls from leaves to roots. However, a local increase in latex total solid content (LILTSC) was identified near the tapping cut of rubber trees. To understand the mechanism of ethephon-stimulated latex yield, the formation and ethephon (an ethylene releaser) alleviation of the LILTSC near the tapping cut were examined. It was found that the LILTSC near the tapping cut of a tapped rubber tree was caused by the tapping-accelerated rubber biosynthesis which began following the first tapping and became significant after the fourth tapping. Ethephon stimulation markedly reduced the LILTSC. The latex yield change pattern upon ethephon stimulation was associated with the kinetic change of LILTSC and the decomposition dynamic of ethephon into ethylene. Once the LILTSC was reduced by ethylene release upon ethephon stimulation, the latex yield increased; however, when the ethylene release upon ethephon stimulation receded, the LILTSC was restored and the effect of ethephon stimulation dissipated. The reduction of LILTSC by ethephon stimulation could be ascribed to the translocation property of ethylene in plants and its regulation of aquaporins. Because maximum ethylene release upon tapping-cut-ethephon-application occured close to the tapping cut, the aquaporins were more up-regulated in this region, leading to a reduction of the LILTSC and an increase in latex yield. All these results suggest that the LILTSC near the tapping cut was caused by tapping; the ethephon-induced aquaporin up-regulation and LILTSC reduction are involved in the mechanism of ethephon-promoted latex yield.

Monitoring the total phenolic/antioxidant levels in wine using flow injection anaylsis with acidic potassium permanganate chemiluminescence detection

Flow injection methodology is described for the estimation of the total phenolic content of wine using acidic potassium permanganate chemiluminescence detection. Selected simple phenolic compounds including quercetin, rutin, catechin, epicatechin, ferulic acid, caffeic acid, gallic acid, 4-hydroxycinnamic acid and vanillin elicited analytically useful chemiluminescence with detection limits ranging between 4×10⁻¹⁰ and 7×10⁻⁷ M. A comparison between the chemiluminescence methodology and other total phenol/antioxidant assays, used by the food and beverage industry, resulted in a good correlation. The chemiluminescence detection was found to be selective with minimal interferences being observed from the non-phenolic components in wine. Analysis of 12 different wines showed that the chemiluminescence method was a rapid way to estimate their antioxidant or total phenolic content.

Seasonal variations of lipid content and composition in perna viridis

The total lipid content, composition of main lipid classes, composition of sterols and composition of fatty acids in the main glycerolipids of Perna viridis were analyzed through four seasons using TLC-FID and GLC. Mussel samples were collected during different seasons between 2003 and 2004 from Shengsi Island, Zhejiang Province, China and stored frozen prior to freeze-drying and lipid extraction. Ten grams of dried mussel powder of each season were analyzed. Total lipid content ranged from 14.5 g/100 g in spring month to 7.8 g/100 g dried mussel powder in autumn month. The predominant lipid in spring month was triacylglycerol (TAG), however, in the other three seasons the phospholipids (PL) was the main lipid class. The most abundant fatty acid in TAG, PL and phosphatidylcholine (PC) was 16:0, with the summer samples having the highest proportion (24-30% of total fatty acid) and winter the lowest (14-22%). In phosphatidylethanolamine (PE), the spring samples had the highest proportions of 16:0. The predominant polyunsaturated fatty acids (PUFA) were 22:6n-3 and 20:5n-3 in TAG, PL, PE and PC (25-40%). The proportions of 22:6n-3 and 20:5n-3 were higher in spring than in other seasons in PL and PE. There were nine sterols identified, with cholesterol being the predominant sterol, and other main ones were desmostersol/brassicasterol and 24-methylenecholesterol. Proportions of other fatty acids in different lipid fractions and the sterol compositions as well also varied seasonally. There were subject to the seasonal variations. Differences in lipid content and composition, fatty acid composition in different lipid fractions may be caused by multiple factors such as lifecycle, sex, variation of plankton in different seasons and temperature, which could influence physiological activities and metabolism.

Seasonal variations of total lipid and fatty acid contents in muscle, gonad and digestive glands of farmed jade tiger hybrid abalone in Australia

The lipid and fatty acid (FA) contents of muscle, gonad and digestive glands (DG) of Jade Tiger hybrid abalone were studied over the four seasons. Higher contents of total lipid and saturated fatty acids (SFA) were found in summer from muscle. For gonad the higher total lipid content was found in summer and spring whereas the SFA content peaked in summer only. For DG the higher contents of total lipid and SFA were recorded in all seasons except autumn. Winter samples showed significantly higher content of PUFA in all three types of tissue. High contents of eicosapentaenoic acid (EPA, 20:5 n−3), docosapentaenoic acid (DPA, 22:5 n−3) and docosahexaenoic acid (DHA, 22:6 n−3) were recorded in winter from muscle, although no marked variations were observed from gonad. For DG the high content of DHA was also observed in winter whilst EPA and DPA maintained high levels in all seasons except summer.

Changes in oil content, lipid class and fatty acid composition of the microalga Chaetoceros calcitrans over different phases of batch culture

 Abstract
The diatom Chaetoceros calcitrans is a microalgal species used as food for larva in aquaculture for many species worldwide. Chaetoceros calcitrans is an important source of omega 3 long chain (C ≥ 20) polyunsaturated fatty acids (n-3 LC PUFA), chiefly eicosapentaenoic acid (EPA, 20:5n-3). This article reports lipid content, lipid class composition and fatty acid profiles of each lipid class during the growth cycle of batch cultures of C. calcitrans. Total lipid content and the concentration of neutral lipid were highest in the late stationary growth phase (day 12). However, the amount of EPA was highest during the logarithmic growth phase (1.24 pg/cell on day 4). EPA was initially concentrated in the glycolipid fraction, but this fraction decreased during logarithmic growth, coinciding with the increase in neutral lipid. Docosahexaenoic acid (22:6n-3, DHA) (0.91 pg/cell) was reported as a major fatty acid (>10 mg/100 g) in all lipid classes on day 1. DHA was depleted quickly from the neutral lipid and glycolipid classes, but at a slower rate from the polar lipid fraction. This work confirms that C. calcitrans is a good source of lipid, in particular EPA, for larval and adult filter feeders in aquaculture.

Effect of sorghum flour addition on resistant starch content, phenolic profile and antioxidant capacity of durum wheat pasta

Foods containing elevated levels of health functional components such as resistant starch and polyphenolic antioxidants may have beneficial effects on human health. Pasta incorporating either red sorghum flour (RSF) or white sorghum flour (WSF) each at 20%, 30% and 40% substitution of durum wheat semolina (DWS) was prepared and compared to pasta made from 100% DWS (control) for content of starch fractions, phenolic profile and antioxidant capacity, before and after cooking. Total, digestible and resistant starch contents were determined by the AOAC method; individual phenolic acids and anthocyanins by reverse phase-HPLC analysis; total phenolic content by the Folin–Ciocalteu method and antioxidant capacity by the ABTS assay. The addition of both RSF and WSF increased the resistant starch content, bound phenolic acids, total phenolic content and antioxidant capacity at all incorporation levels compared to the control pasta; while free phenolic acids and anthocyanins were higher in the RSF-containing pasta only. Cooking did not change the resistant starch content of any of the pasta formulations. Cooking did however decrease the free phenolic acids, anthocyanins, total phenolic content and antioxidant capacity and increased the bound phenolic acids of the sorghum-containing pastas. The study suggests that these sorghum flours may be very useful for the preparation of pasta with increased levels of resistant starch and polyphenolic antioxidants.

Endurance training in obese humans improves glucose tolerance and mitochondrial fatty acid oxidation and alters muscle lipid content

Muscle fatty acid (FA) metabolism is impaired in obesity and insulin resistance, reflected by reduced rates of FA oxidation and accumulation of lipids. It has been suggested that interventions that increase FA oxidation may enhance insulin action by reducing these lipid pools. Here, we examined the effect of endurance training on rates of mitochondrial FA oxidation, the activity of carnitine palmitoyltransferase I (CPT I), and the lipid content in muscle of obese individuals and related these to measures of glucose tolerance. Nine obese subjects completed 8 wk of moderate-intensity endurance training, and muscle biopsies were obtained before and after training. Training significantly improved glucose tolerance, with a reduction in the area under the curve for glucose (P< 0.05) and insulin (P = 0.01) during an oral glucose tolerance test. CPT I activity increased 250% (P = 0.001) with training and became less sensitive to inhibition by malonyl-CoA. This was associated with an increase in mitochondrial FA oxidation (+120%, P < 0.001). Training had no effect on muscle triacylglycerol content; however, there was a trend for training to reduce both the total diacylglcyerol (DAG) content (−15%, P = 0.06) and the saturated DAG-FA species (−27%, P = 0.06). Training reduced both total ceramide content (−42%, P = 0.01) and the saturated ceramide species (−32%, P < 0.05). These findings suggest that the improved capacity for mitochondrial FA uptake and oxidation leads not only to a reduction in muscle lipid content but also a to change in the saturation status of lipids, which may, at least in part, provide a mechanism for the enhanced insulin action observed with endurance training in obese individuals.

Disassociation of muscle triglyceride content and insulin sensitivity after exercise training in patients with Type 2 diabetes

Aim/hypothesis. We determined the effect of exercise training on insulin sensitivity and muscle lipids (triglyceride [TGm] and long-chain fatty acyl CoA [LCACoA] concentration) in patients with Type 2 diabetes. Methods. Seven patients with Type 2 diabetes and six healthy control subjects who were matched for age, BMI, % body fat and VO2peak participated in a 3 days per week training program for 8 weeks. Insulin sensitivity was determined pre- and post-training during a 120 min euglycaemic- hyperinsulinaemic clamp and muscle biopsies were obtained before and after each clamp. Oxidative enzyme activities [citrate synthase (CS), β-hydroxy-acyl- CoA (β-HAD)] and TGm were determined from basal muscle samples pre- and post training, while total LCACoA content was measured in samples obtained before and after insulin-stimulation, pre- and post training. Results. The training-induced increase in VO2peak (∼20%, p<0.01) was similar in both groups. Compared with control subjects, insulin sensitivity was lower in the diabetic patients before and after training (∼60%; p<0.05), but was increased to the same extent in both groups with training (∼30%; p<0.01). TGm was increased in patients with Type 2 diabetes (170%; p<0.05) before, but was normalized to levels observed in control subjects after training. Basal LCACoA content was similar between groups and was unaltered by training. Insulin-stimulation had no detectable effect on LCACoA content. CS and β-HAD activity were increased to the same extent in both groups in response to training (p<0.001). Conclusion/interpretation. We conclude that the enhanced insulin sensitivity observed after short-term exercise training was associated with a marked decrease in TGm content in patients with Type 2 diabetes. However, despite the normalization of TGm to levels observed in healthy individuals, insulin resistance was not completely reversed in the diabetic patients.

The microstructure and physicochemical properties of probiotic buffalo yoghurt during fermentation and storage: a comparison with bovine yoghurt

The physicochemical and rheological properties of yoghurt made from unstandardised unhomogenised buffalo milk were investigated during fermentation and 28 days of storage and compared to the properties of yoghurt made from homogenised fortified bovine milk. A number of differences observed in the gel network can be linked to differences in milk composition. The microstructure of buffalo yoghurt, as assessed by confocal laser scanning microscopy (CLSM) and cryo scanning electron microscopy (cryo-SEM), was interrupted by large fat globules and featured more serum pores. These fat globules have a lower surface area and bind less protein than the homogenised fat globules in bovine milk. These microstructural differences likely lead to the higher syneresis observed for buffalo yoghurt with an increase from 17.4 % (w/w) to 19.7 % (w/w) in the weight of whey generated at days 1 and 28 of the storage. The higher concentration of total calcium in buffalo milk resulted in the release of more ionic calcium during fermentation. Gelation was also slower but the strength of the two gels was similar due to similar protein and total solids concentrations. Buffalo yoghurt was more viscous, less able to recover from deformation and less Newtonian than bovine yoghurt with a thixotropy of 3,035 Pa.s-1 measured for buffalo yoghurt at the end of the storage, at least four times higher than the thixotropy of bovine yoghurt. While the titratable acidity, lactose consumption and changes in organic acid concentrations were similar, differences were recorded in the viability of probiotic bacteria with a lower viability of Lactobacillus acidophilus of 5.17 log (CFU/g) recorded for buffalo yoghurt at day 28 of the storage. Our results show that factors other than the total solids content and protein concentration of milk affect the structural properties of yoghurt. They also illustrate the physicochemical reasons why buffalo and bovine yoghurt are reported to have different sensory properties and provide insight into how compositional changes can be used to alter the microstructure and properties of dairy products. © 2013 Springer Science+Business Media New York.

Creatine and the creatine transporter: a review

The cellular role of creatine (Cr) and Cr phosphate (CrP) has been studied extensively in neural, cardiac and skeletal muscle. Several studies have demonstrated that alterations in the cellular total Cr (Cr + CrP) concentration in these tissues can produce marked functional and/or structural change. The primary aim of this review was to critically evaluate the literature that has examined the regulation of cellular total Cr content. In particular, the review focuses on the regulation of the activity and gene expression of the Cr transporter (CreaT), which is primarily responsible for cellular Cr uptake. Two CreaT genes (CreaT1 and CreaT2) have been identified and their chromosomal location and DNA sequencing have been completed. From these data, putative structures of the CreaT proteins have been formulated. Transcription products of the CreaT2 gene are expressed exclusively in the testes, whereas CreaT1 transcripts are found in a variety of tissues. Recent research has measured the expression of the CreaT1 protein in several tissues including neural, cardiac and skeletal muscle. There is very little information available about the factors regulating CreaT gene expression. There is some evidence that suggests the intracellular Cr concentration may be involved in the regulatory process but there is much more to learn before this process is understood. The activity of the CreaT protein is controlled by many factors. These include substrate concentration, transmembrane Na+ gradients, cellular location, and various hormones. It is also likely that transporter activity is influenced by its phosphorylation state and by its interaction with other plasma membrane proteins. The extent of CreaT protein glycosylation may vary within cells, the functional significance of which remains unclear.

Emerging technologies in building envelopes

Advancements in engineering and space technology are increasingly finding application in buildings. Building envelopes are utilising components of high-technological solutions resulting in better visibility, greater light transmission, increased energy generation and storage capacity, improved shading and ventilation and integration with the external environment. This report summarises several technological advancements and suggests forthcoming directions for building envelope design. Many of the technologies presented here have been invented and developed in Australia, yet are not commonly used by the building construction industry.

Emerging technologies in ventilation

Innovation in ventilation systems is becoming an increasingly popular and targeted topic of architectural discourse. Architects, consultants and contractors are introducing new products and proposing new systems, subject to client requests for an environmentally responsive architecture. The authors, in compiling the research for this guide, experienced a large increase in Australian constructed buildings that focused specifically on ventilation strategies and systems. This note presents and discusses the underlying principles of different ventilation techniques. Applications of specific ventilation techniques are demonstrated through building examples constructed in Australia as well as overseas. Although a particular building design may demonstrate several ventilation concepts simultaneously, this note illustrates the most dominant ventilation features in each example.

Effect of creatine supplementation on housekeeping genes in human skeletal muscle using real-time RT-PCR

The present study examined the validity and reliability of measuring the expression of various genes in human skeletal muscle using quantitative real-time RT-PCR on a GeneAmp 5700 sequence detection system with SYBR Green 1 chemistry. In addition, the validity of using some of these genes as endogenous controls (i.e., housekeeping genes) when human skeletal muscle was exposed to elevated total creatine levels and exercise was also examined. For all except 28S, linear relationships between the logarithm of the starting RNA concentrations and the cycle threshold (C_T) values were established for ß-actin, ß2-microglobulin (ß₂M), cyclophilin (CYC), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a linear response between C_T values and the logarithm of a given amount of starting cDNA for all the genes tested. The overall intra-assay coefficient of variance for these genes was 1.3% and 21% for raw C_T values and the linear value of 2^-C_T, respectively. Interassay variability was 2.3% for raw C_T values and 34% for the linear value of 2^-C_T. We also examined the expression of various housekeeping genes in human skeletal muscle at days 0, 1, and 5 following oral supplementation with either creatine or a placebo employing a double-blind crossover study design. Treatments were separated by a 5-wk washout period. Immediately following each muscle sampling, subjects performed two 30-s all-out bouts on a cycle ergometer. Creatine supplementation increased (P < 0.05) muscle total creatine content above placebo levels; however, there were no changes (P > 0.05) in C_T values across the supplementation periods for any of the genes. Nevertheless, 95% confidence intervals showed that GAPDH was variable, whereas ß-actin, ß₂M, and CYC were the least varying genes. Normalization of the data to these housekeeping genes revealed variable behavior for ß₂M with more stable expressions for both ß-actin and CYC. We conclude that, using real-time RT-PCR, ß-actin or CYC may be used as housekeeping genes to study gene expression in human muscle in experiments employing short-term creatine supplementation combined with high-intensity exercise.