Identification of salivary N-glycoproteins and measurement of glycosylation site occupancy by boronate glycoprotein enrichment and liquid chromatography/electrospray ionization tandem mass spectrometry

RATIONALE Diseases including cancer and congenital disorders of glycosylation have been associated with changes in the site-specific extent of protein glycosylation. Saliva can be non-invasively sampled and is rich in glycoproteins, giving it the potential to be a useful biofluid for the discovery and detection of disease biomarkers associated with changes in glycosylation. METHODS Saliva was collected from healthy individuals and glycoproteins were enriched using phenylboronic acid based glycoprotein enrichment resin. Proteins were deglycosylated with peptide-N-glycosidase F and digested with AspN or trypsin. Desalted peptides and deglycosylated peptides were separated by reversed-phase liquid chromatography and detected with on-line electrospray ionization quadrupole-time-of-flight mass spectrometry using a 5600 TripleTof instrument. Site-specific glycosylation occupancy was semi-quantitatively determined from the abundance of deglycosylated and nonglycosylated versions of each given peptide. RESULTS Glycoprotein enrichment identified 67 independent glycosylation sites from 24 unique proteins, a 3.9-fold increase in the number of glycosylation sites identified. Enrichment of glycoproteins rather than glycopeptides allowed detection of both deglycosylated and nonglycosylated versions of each peptide, and thereby robust measurement of site-specific occupancy at 21 asparagines. Healthy individuals showed limited biological variability in occupancy, with partially modified sites having characteristics consistent with inefficient glycosylation by oligosaccharyltransferase. Inclusion of negative controls without enzymatic deglycosylation controlled for spontaneous chemical deamidation, and identified asparagines previously incorrectly annotated as glycosylated. CONCLUSIONS We developed a sample preparation and mass spectrometry detection strategy for rapid and efficient measurement of site-specific glycosylation occupancy on diverse salivary glycoproteins suitable for biomarker discovery and detection of changes in glycosylation occupancy in human disease.

Multiphase porous media model for heat and mass transfer during drying of agricultural products

Modelling of food processing is complex because it involves sophisticated material and transport phenomena. Most of the agricultural products such fruits and vegetables are hygroscopic porous media containing free water, bound water, gas and solid matrix. Considering all phase in modelling is still not developed. In this article, a comprehensive porous media model for drying has been developed considering bound water, free water separately, as well as water vapour and air. Free water transport was considered as diffusion, pressure driven and evaporation. Bound water assumed to be converted to free water due to concentration difference and also can diffuse. Binary diffusion between water vapour and air was considered. Since, the model is fundamental physics based it can be applied to any drying applications and other food processing where heat and mass transfer takes place in porous media with significant evaporation and other phase change.

Characterization of acyl chain position in unsaturated phosphatidylcholines using differential mobility-mass spectrometry

Glycerophospholipids (GPs) that differ in the relative position of the two fatty acyl chains on the glycerol backbone (i.e., sn-positional isomers) can have distinct physicochemical properties. The unambiguous assignment of acyl chain position to an individual GP represents a significant analytical challenge. Here we describe a workflow where phosphatidylcholines (PCs) are subjected to ESI for characterization by a combination of differential mobility spectrometry and MS (DMS-MS). When infused as a mixture, ions formed from silver adduction of each phospholipid isomer {e.g., [PC (16:0/18:1) + Ag]+ and [PC (18:1/16:0) + Ag]+} are transmitted through the DMS device at discrete compensation voltages. Varying their relative amounts allows facile and unambiguous assignment of the sn-positions of the fatty acyl chains for each isomer. Integration of the well-resolved ion populations provides a rapid method (< 3 min) for relative quantification of these lipid isomers. The DMS-MS results show excellent agreement with established, but time-consuming, enzymatic approaches and also provide superior accuracy to methods that rely on MS alone. The advantages of this DMS-MS method in identification and quantification of GP isomer populations is demonstrated by direct analysis of complex biological extracts without any prior fractionation.

Studies of Brisbane municipal water quality using Inductively Coupled Plasma - Mass Spectrometry and Chemometrics

This research established innovative methods and a predictive model to evaluate water quality using the trace element and heavy metal concentrations of drinking water from the greater Brisbane area. Significantly, the combined use of Inductively Coupled Plasma - Mass Spectrometry and Chemometrics can be used worldwide to provide comprehensive, rapid and affordable analyses of elements in drinking water that can have a considerable impact on human health.

A high performance liquid chromatography and electrospray ionization mass spectrometry method for the analysis of the natural medicine, Forsythia suspensa

Samples of Forsythia suspensa from raw (Laoqiao) and ripe (Qingqiao) fruit were analyzed with the use of HPLC-DAD and the EIS-MS techniques. Seventeen peaks were detected, and of these, twelve were identified. Most were related to the glucopyranoside molecular fragment. Samples collected from three geographical areas (Shanxi, Henan and Shandong Provinces), were discriminated with the use of hierarchical clustering analysis (HCA), discriminant analysis (DA), and principal component analysis (PCA) models, but only PCA was able to provide further information about the relationships between objects and loadings; eight peaks were related to the provinces of sample origin. The supervised classification models-K-nearest neighbor (KNN), least squares support vector machines (LS-SVM), and counter propagation artificial neural network (CP-ANN) methods, indicated successful classification but KNN produced 100% classification rate. Thus, the fruit were discriminated on the basis of their places of origin.

Extinction risk analysis of exploited green turtle stocks in the Indo-Pacific

In parts of the Indo-Pacific, large-scale exploitation of the green turtle Chelonia mydas continues to pose a serious threat to the persistence of this species; yet very few studies have assessed the pattern and extent of the impact of such harvests. We used demographic and genetic data in an age-based model to investigate the viability of an exploited green turtle stock from Aru, south-east Indonesia. We found that populations are decreasing under current exploitation pressures. The effects of increasingly severe exploitation activities at foraging and nesting habitat varied depending on the migratory patterns of the stock. Our model predicted a rapid decline of the Aru stock in Indonesia under local exploitation pressure and a shift in the genetic composition of the stock. We used the model to investigate the influence of different types of conservation actions on the persistence of the Aru stock. The results show that local management actions such as nest protection and reducing harvests of adult nesting and foraging turtles can have considerable conservation outcomes and result in the long-term persistence of genetically distinct management units. © 2010 The Authors. Animal Conservation © 2010 The Zoological Society of London.

Extinction risk in cloud forest fragments under climate change and habitat loss

Aim: To quantify the consequences of major threats to biodiversity, such as climate and land-use change, it is important to use explicit measures of species persistence, such as extinction risk. The extinction risk of metapopulations can be approximated through simple models, providing a regional snapshot of the extinction probability of a species. We evaluated the extinction risk of three species under different climate change scenarios in three different regions of the Mexican cloud forest, a highly fragmented habitat that is particularly vulnerable to climate change. Location: Cloud forests in Mexico. Methods: Using Maxent, we estimated the potential distribution of cloud forest for three different time horizons (2030, 2050 and 2080) and their overlap with protected areas. Then, we calculated the extinction risk of three contrasting vertebrate species for two scenarios: (1) climate change only (all suitable areas of cloud forest through time) and (2) climate and land-use change (only suitable areas within a currently protected area), using an explicit patch-occupancy approximation model and calculating the joint probability of all populations becoming extinct when the number of remaining patches was less than five. Results: Our results show that the extent of environmentally suitable areas for cloud forest in Mexico will sharply decline in the next 70 years. We discovered that if all habitat outside protected areas is transformed, then only species with small area requirements are likely to persist. With habitat loss through climate change only, high dispersal rates are sufficient for persistence, but this requires protection of all remaining cloud forest areas. Main conclusions: Even if high dispersal rates mitigate the extinction risk of species due to climate change, the synergistic impacts of changing climate and land use further threaten the persistence of species with higher area requirements. Our approach for assessing the impacts of threats on biodiversity is particularly useful when there is little time or data for detailed population viability analyses. © 2013 John Wiley & Sons Ltd.

Paying the extinction debt: Woodland birds in the Mount Lofty Ranges, South Australia

Approximately 90% of the original woodlands of the Mount Lofty Ranges of South Australia has been cleared, modified or fragmented, most severely in the last 60 years, and affecting the avifauna dependent on native vegetation. This study identifies which woodland-dependent species are still declining in two different habitats, Pink GumBlue Gum woodland and Stringybark woodland. We analyse the Mount Lofty Ranges Woodland Bird Long-Term Monitoring Dataset for 1999-2007, to look for changes in abundance of 59 species. We use logistic regression of prevalence on lists in a Bayesian framework, and List Length Analysis to control for variation in detectability. Compared with Reporting Rate Analysis, a more traditional approach, List Length Analysis provides tighter confidence intervals by accounting for changing detectability. Several common species were declining significantly. Increasers were generally large-bodied generalists. Many birds have already disappeared from this modified and naturally isolated woodland island, and our results suggest that more specialist insectivores are likely to follow. The Mount Lofty Ranges can be regarded as a 'canary landscape' for temperate woodlands elsewhere in Australia without immediate action their bird communities are likely to follow the trajectory of the Mount Lofty Ranges avifauna. Alternatively, with extensive habitat restoration and management, we could avoid paying the extinction debt. © Royal Australasian Ornithologists Union 2011.

Prospective ten-month exercise intervention in premenarcheal girls: Positive effects on bone and lean mass

Enhancement of bone mineral acquisition during growth may be a useful preventive strategy against osteoporosis. The aim of this study was to explore the lean mass, strength, and bone mineral response to a 10-month, high-impact, strength-building exercise program in 71 premenarcheal girls, aged 9–10 years. Lean body mass, total body (TB), lumbar spine (LS), proximal femur (PF), and femoral neck (FN) bone mineral were measured using the Hologic QDR 2000+ bone densitometer. Strength was assessed using a grip dynamometer and the Cybex isokinetic dynamometer (Cybex II). At baseline, no significant difference in body composition, pubertal development, calcium intake, physical activity, strength, or bone mineral existed between groups. At completion, there were again no differences in height, total body mass, pubertal development, calcium intake, or external physical activity. In contrast, the exercise group gained significantly more lean mass, less body fat content, greater shoulder, knee and grip strength, and greater TB, LS, PF, and FN BMD (exercise: TB 3.5%, LS 4.8%, PF 4.5%, and FN 12.0%) compared with the controls (controls: TB 1.2%, LS 1.2%, PF 1.3%, and FN 1.7%). TB bone mineral content (BMC), LS BMC, PF BMC, FN BMC, LS bone mineral apparent density (BMAD), and FN bone area also increased at a significantly greater rate in the exercise group compared with the controls. In multiple regression analysis, change in lean mass was the primary determinant of TB, FN, PF, and LS BMD accrual. Although a large proportion of bone mineral accrual in the premenarcheal skeleton was related to growth, an osteogenic effect was associated with exercise. These results suggest that high-impact, strength building exercise is beneficial for premenarcheal strength, lean mass gains, and bone mineral acquisition.

Technological adoption and use after mass market displacement: The case of the LP record

This paper investigates the reasons why some technologies, defying general expectations and the established models of technological change, may not disappear from the market after having been displaced from their once-dominant status. Our point of departure is that the established models of technological change are not suitable to explain this as they predominantly focus on technological dominance, giving attention to the technologies that display highest performance levels and gain greatest market share. And yet, technological landscapes are rife with technological designs that do not fulfil these conditions. Using the LP record as an empirical case, we propose that the central mechanism at play in the continuing market presence of once-dominant technologies is the recasting of their technological features from the functional-utilitarian to the aesthetic realm, with an additional element concerning communal interaction among users. The findings that emerge from our quantitative textual analysis of over 200,000 posts on a prominent online LP-related discussion forum (between 2002 and 2010) also suggest that the post-dominance technology adopters and users appear to share many key characteristics with the earliest adopters of new technologies, rather than with late-stage adopters which precede them.

Speed-based diagnostics of aerodynamic and mass imbalance in large wind turbines

Any kind of imbalance in the operation of a wind turbine has adverse effect on the downstream torsional components as well as tower structure. It is crucial to detect imbalance at its very inception. The identification of the type of imbalance is also required so that appropriate measures of fault accommodation can be performed in the control system. In particular, it is important to distinguish between mass and aerodynamic imbalance. While the former is gradually caused by a structural anomaly (e.g. ice deposition, moisture accumulation inside blade), the latter is generally associated to a fault in the pitch control system. This paper proposes a technique for the detection and identification of imbalance fault in large scale wind turbines. Unlike most other existing method it requires only the rotor speed signal which is readily available in existing turbines. Signature frequencies have been proposed in this work to identify imbalance type based on their physical phenomenology. The performance of this technique has been evaluated by simulations using an existing benchmark model. The effectiveness of the proposed method has been confirmed by the simulation results.