Top-down proteomic analysis by MALDI-TOF profiling: Concentration-independent biomarkers

Although numerous protein biomarkers have been correlated with advanced disease states, no new clinical assays have been developed. Goals often anticipate disease-specific protein changes that exceed values among healthy individuals, a property common to acute phase reactants. This review considers somewhat different approaches. It focuses on intact protein isoform ratios that present a biomarker without change in the total concentration of the protein. These will seldom be detected by peptide level analysis or by most antibody-based assays. For example, application of an inexpensive method to large sample groups resulted in observation of several polymorphisms, including the first structural polymorphism of apolipoprotein C1. Isoform distribution of this protein was altered and was eventually linked to increased obesity. Numerous other protein isoforms included C- and N-terminal proteolysis, changes of glycoisoform ratios and certain types of sulfhydryl oxidation. While many of these gave excellent statistical correlation with advanced disease, clinical utility was not apparent. More important may be that protein isoform ratios were very stable in each individual. Diagnosis by longitudinal analysis of the same individual might increase sensitivity of protein biomarkers by 20-fold or more. Protein changes that exceed the range of values found among healthy individuals may be uncommon.     

Triton X-100 pretreatment of LR-white thin sections improves immunofluorescence specificity and intensity

The staining of intracellular antigenic sites in postembedded samples is a challenging problem. Deterioration of antigenicity and limited antibody accessibility to the antigen are commonly encountered on account of processing steps. In this study preservation of the antigen was achieved by fixing the tissues with mild fixatives, performing partial dehydration, and embedding in a low crosslinked hydrophilic acrylic resin, LR-White. Permeabilization of cell membranes with Triton X-100 is well documented but can affect some antigen conformations. We tested the effect of Triton X-100 on the ED1 antigen present in the lysosomal membrane of the macrophage in cell culture. The ED1 antigen in the lysosome was resistant to extraction by Triton X-100. Interestingly pretreating the LR-White sections of macrophage pellets with Triton X-100 improved the staining intensity of ED1. The most intense and clear specific fluorescent staining was observed when sections were pretreated with 0.2% Triton X-100 for 2 min. Longer exposure of sections to 0.2% Triton or 2 min exposure to 2% Triton lead to reduced ED1 labeling. SEM observations indicated that the detergent extracted a component from the cells and not the resin and was determined to be lipid. This novel technique could be applied in many research areas where postembedding fluorescent immunolabeling with higher labeling intensity is desired.     

A model for the prediction of the forces at the glenohumeral joint

The design, validation, and application of a three-dimensional musculoskeletal model of the upper limb is presented here, for the study of joint, muscle, and ligament forces during selected tasks of daily living. Much of the model was constructed using data gathered from real cadavers, including the extensive 'visible human' data set, together with information gathered from published post mortem studies on cadavers. Validation was carried out through comparison of model outcomes with available data in the literature. The model was then used to examine the glenohumeral joint loading during ten previously selected activities of daily living. The predicted glenohumeral joint contact forces during everyday activities were found to range from 23 to 75 per cent body weight. A linear regression method for estimating the joint load from externally measured kinetics was developed, giving a 95 per cent confidence interval in predicted joint forces of 8 per cent body weight. The model presented here, namely the Newcastle shoulder model, provides a basis for future studies into biomechanical problems relating to the understanding of musculoskeletal pathology and implant design and performance.     

The nutritive value of protein isolates and fibres from meat industry by-products

Protein isolates extracted from rumen, lung and plasma by alkaline solubilisation were spun into protein fibres and were evaluated for protein quality by rat feeding trials and by amino acid analyses. The net protein utilisation (NPU) of the fibres ranged from 53·0 to 76·9 for plasma and rumen fibres respectively, methionine plus cystine and valine being limiting amino acids. Lysine was not found to be a limiting amino acid in any sample and lysine availability was high in isolates and fell only slightly on spinning. There was a marked discrepancy between chemical score and NPU for rumen isolate and it is postulated that an anti-nutritional factor, possibly a trypsin inhibitor, normally present in bovine organs, could be active in the isolate by lowering methionine availability to the animal and decreasing NPU. Spinning the proteins, however, either destroys the inhibitor or decreases its concentration since NPU and chemical score become equal.     

The effects of alkaline hydrogen peroxide treatment on the nutritional value of tripe

Samples of tripe obtained after commercial processing with alkaline hydrogen peroxide were compared with raw tripe for nutritional quality. A large number (108) of commercial tripe samples were found to vary markedly in their protein content from 8·4 to 19·3%. The moisture content was negatively correlated with the pH of the tripe samples. However, the amino acid analyses and rat bioassays indicated that the nutritional value of the protein was not significantly decreased by the processing. Oxidation of sulphur amino acids and formation of lysino-alanine were not evident. Thiamine was completely destroyed by the processing.     

The effect of scale manipulations on validity: targetting frequency rating scales for anticipated performance levels

A common problem with standard five-point frequency rating scales is their inability to differentiate between objects within a relatively narrow band of the rating dimension. Two alternatives for increasing a scale's ability to reflect existing differences are: increasing the number of positions on the rating scale, or packing the rating scale with quantifiers from a particular portion of the frequency dimension. In this study, three types of rating scale - a standard five-point balanced scale, a longer nine-point balanced scale, and a five-point packed scale - were used to rate two videotaped samples of behaviour, one displaying performance levels from 10% to 100% and the other from 70% to 90%. Each subject's ratings were correlated with true performance levels in each sample of behaviour as a measure of validity. Results showed that for ratings of the wide range of behaviour all three types of scales provided average correlations between the ratings and actual frequencies of the event which exceeded 0.90, with the longer nine-point scale yielding a significantly higher mean correlation than the other two scales. For ratings of the narrow performance range, the nine-point scale provided the highest correlation with the actual frequencies, followed by the packed scale and the standard five-point balanced scale. All differences were significant. Findings suggest that increasing the number of scale positions can significantly increase the validity of ratings obtained. Also, though to a lesser degree, validity of ratings may be enhanced in a shorter scale by using quantifiers from the portion of the frequency continuum where performance are anticipated to lie.     

Determination of dairy wastewater treatability by bio-trickling filter packed with lava rocks - case study PEGAH dairy factory

This paper investigates the effectiveness of a biological trickling filter for the treatment of wastewaters produced by a company manufacturing dairy products. First a bio-trickling column with a height of 150 cm was packed with lava rocks from north mountain of Tehran. It operates with the recirculation of liquid through the packing. In order to startup the pilot scale, steady state condition was gained by pumping activated sludge and dairy wastewater for 23 days. Afterwards, dairy wastewater was added to liquid tank for treatment. Hydraulic retention time (HRT) of treatment decreases from 5 days to 1 day then at HRT of 12, 8, 7, 6 and 4 h. Results show that the average chemical oxygen demand (COD) decreased from 2,750 to 98 mg/L at HRT of 7 h and efficiency of TKN removal was more than 70%. The microorganisms developed in the bio-trickling filter were able to efficiently remove COD levels up to 2,750 mg/L, under aerobic conditions at pH values between 6.8 and 7.2 under low temperature condition between 10 and 13 °C.     

Implications to stormwater management as a result of lot scale rainwater tank systems: a case study in Western Sydney, Australia

Rainwater tanks are increasingly adopted in Australia to reduce potable water demand and are perceived to reduce the volume of stormwater discharge from developments. This paper investigates the water balance of rainwater tanks, in particular the possible impacts these tanks could have in controlling the stormwater discharge volume. The study collected water quantity data from two sites in the Hawkesbury City Council area, New South Wales, Australia and utilised the collected data in a simple water balance model to assess the effectiveness of rainwater tanks in reducing the stormwater discharge volume. The results indicate that a significant reduction in discharge volume from a lot scale development can be achieved if the rainwater tank is connected to multiple end-uses, but is minimal when using irrigation alone. In addition, the commonly used volumetric runoff coefficient of 0.9 was found to over-estimate the runoff from the roof areas and to thereby under-estimate the available volume within the rainwater tanks for retention or detention. Also, sole reliance on the water in the rainwater tanks can make the users aware of their water use pattern and water availability, resulting in significant reductions in water use as the supply dwindles, through self-imposed water restrictions.     

Measurement and mathematical modelling of nutrient level and water quality parameters

Physico-chemical water quality parameters and nutrient levels such as water temperature, turbidity, saturated oxygen, dissolved oxygen, pH, chlorophyll-a, salinity, conductivity, total nitrogen and total phosphorus, were measured from April to September 2011 in the Karaj dam area, Iran. Total nitrogen in water was modelled using an artificial neural network system. In the proposed system, water temperature, depth, saturated oxygen, dissolved oxygen, pH, chlorophyll-a, salinity, turbidity and conductivity were considered as input data, and the total nitrogen in water was considered as output. The weights and biases for various systems were obtained by the quick propagation, batch back propagation, incremental back propagation, genetic and Levenberg-Marquardt algorithms. The proposed system uses 144 experimental data points; 70% of the experimental data are randomly selected for training the network and 30% of the data are used for testing. The best network topology was obtained as (9-5-1) using the quick propagation method with tangent transform function. The average absolute deviation percentages (AAD%) are 2.329 and 2.301 for training and testing processes, respectively. It is emphasized that the results of the artificial neural network (ANN) model are compatible with the experimental data.