Inhibition of protease activity. Part 1. The effect on tenderness and indicators of proteolysis in ovine muscle

To examine the effect of particular enzyme groups on tenderness specific cysteine protease inhibitors were injected into muscle early post-mortem. The protease enzyme inhibitor E-64 was injected into the m. longissimus thoracis et lumborum (LTL) on the right side of 12 lamb carcasses within 15 min of death and in another 12 carcasses with the protease inhibitor Z-Phe-Ala-CHN(2). The left LTL (control) was injected with saline (0.25 M NaCl). To create variation in the rate of pH decline alternate carcasses were electrically stimulated (low voltage). The LTL was divided into cranial and caudal portions and aged for 1 or 2 days. Muscle samples at 1 day post-mortem were used for measurement of osmolality and sarcomere length (n=48), and others at 1 and 2 days post-mortem for shear force determination (n=96). The myofibrillar fragmentation index (MFI) was determined on samples taken at pH 6.2 and 1 and 2 days post-mortem (n=144). Other muscle samples were obtained at death, pH 6.2 and 6.0 and then at 1 and 2 days post-mortem (n=215). These samples were used for determination of protein solubility and the concentration of free amino acids. Stimulation caused a faster (P<0.05) decline in pH. There was no effect of stimulation (P>0.05) on shear force values, but injection of inhibitor and ageing both had effects (P<0.001). The inhibitor E-64 prevented any improvement in tenderness with ageing, whereas the inhibitor Z-Phe-Ala-CHN(2) and the control samples showed a similar ageing response. In the latter two treatments there was an average reduction of 1 kg in shear between 1 and 2 days post-mortem, whilst the inhibitor E-64 maintained shear force on average 2 kg higher than control samples. Injection and ageing had an effect on MFI (P<0.001) and there was an interaction (P<0.05) between stimulation and ageing for MFI, such that as stimulated muscle aged the rate of change of MFI was greater. There was an interaction between injection and ageing (P<0.05) for protein solubility such that samples treated with E-64 showed a minimal increase in protein solubility with ageing, whereas in samples treated with Z-Phe-Ala-CHN(2) and the control samples there was a significant increase. There was also an interaction between stimulation and ageing such that between sampling at pH 6.0 and 2 days post-mortem, stimulated muscle exhibited greater solubility (P<0.05). There were no effects (P>0.05) on the concentration of free amino acids. The evidence indicated that the cysteine proteases were responsible for post-mortem proteolysis and tenderisation, in particular the calpains, whereas the cathepsins (B and L) were unlikely to contribute to proteolysis and subsequent tenderisation in meat.     

Eating quality of commercially processed hot boned sheep meat

The aim of this work was to establish consumer perceptions of the eating quality of commercially processed hot boned sheep meat. The eating quality scores for tenderness, flavour, juiciness, and overall liking of grilled m. longissimus thoracis et lumborum (LL) were derived from untrained consumers. The animals used in this experiment were sourced from three different properties and were of various ages, breed, and sex, and had been on various quality pastures. All carcases were subjected to immobilisation, spinal discharge and high voltage electrical stimulation and boned within 2 h of slaughter. All LL samples were frozen after boning according to the system applied at the abattoir and samples kept for consumer assessment and measurement of meat quality. The results showed, on average carcases entered rigor at high temperatures with a pH of 5.95 at 29.3 °C. With an average sarcomere length of 1.68 μm, some shortening was evident and there was a significant difference between lots in sarcomere length (P < 0.05). A large percentage (82%) of samples had a pH greater than 5.8 at the time of freezing. All samples taken for shear force analysis exceeded a recommended threshold for acceptable table meat of 49 N and there was no significant difference between lots (P > 0.05). Only 13.5% of the samples met the ‘good everyday’ requirement following sensory assessment and there were significant differences between lots for eating quality traits (P < 0.05). This work clearly shows that the application of effective electrical stimulation is not sufficient to ensure that hot boned sheep meat will be suitable as a table meat. These findings highlight the need for the inclusion of other intervention techniques.     

Planning a City through ‘Dialogue’: Deliberative Policy-making in Action in Western Australia

Deliberative policy-making is an alternative method of generating new planning strategies based on the public deliberation of citizens and stakeholders. Four ideals of deliberative policy-making espoused by deliberative democracy theorists are identified. Using this framework the ‘Dialogue with the City’ process undertaken by the Western Australian Government to develop the Network City spatial planning strategy for the Perth metropolitan region is examined. While this process went further than most towards realising the ideals of deliberative policy-making, the Government was not able to fully achieve the ideals of inclusive decision-making, equitable decision-making, deliberation toward the common good and consensus decision-making. It is most likely that the deliberative approach to policy-making does not represent a new model for governing; rather, these participation techniques constitute a more sophisticated model for involving citizens and stakeholders in planning.

     

Immunologic Effects of Stereotactic Body Radiotherapy in Dogs with Spontaneous Tumors and the Impact of Intratumoral OX40/TLR Agonist Immunotherapy

Stereotactic body radiotherapy (SBRT) is known to induce important immunologic changes within the tumor microenvironment (TME). However, little is known regarding the early immune responses within the TME in the first few weeks following SBRT. Therefore, we used the canine spontaneous tumor model to investigate TME responses to SBRT, and how local injection of immune modulatory antibodies to OX40 and TLR 3/9 agonists might modify those responses. Pet dogs with spontaneous cancers (melanoma, carcinoma, sarcoma, n = 6 per group) were randomized to treatment with either SBRT or SBRT combined with local immunotherapy. Serial tumor biopsies and serum samples were analyzed for immunologic responses. SBRT alone resulted at two weeks after treatment in increased tumor densities of CD3+ T cells, FoxP3+ Tregs, and CD204+ macrophages, and increased expression of genes associated with immunosuppression. The addition of OX40/TLR3/9 immunotherapy to SBRT resulted in local depletion of Tregs and tumor macrophages and reduced Treg-associated gene expression (FoxP3), suppressed macrophage-associated gene expression (IL-8), and suppressed exhausted T cell-associated gene expression (CTLA4). Increased concentrations of IL-7, IL-15, and IL-18 were observed in serum of animals treated with SBRT and immunotherapy, compared to animals treated with SBRT. A paradoxical decrease in the density of effector CD3+ T cells was observed in tumor tissues that received combined SBRT and immunotherapy as compared to animals treated with SBRT only. In summary, these results obtained in a spontaneous large animal cancer model indicate that addition of OX40/TLR immunotherapy to SBRT modifies important immunological effects both locally and systemically.     

Growth of a human intestinal Desulfovibrio desulfuricans in continuous cultures containing defined populations of saccharolytic and amino acid fermenting bacteria

Ecological and physiological effects of the sulphate-reducing bacterium (SRB) Desulfovibrio desulfuricans on other intestinal organisms were investigated in anaerobic chemostats (dilution rate approximately 0.2 h-1). Reproducible defined bacterial communities were used in these experiments, comprising 14 different saccharolytic and amino acid fermenting species: Bifidobacterium longum, Bif. adolescentis, Bif. pseudolongum, Bif. infantis, Bacteroides thetaiotaomicron, Bact. vulgatus, Lactobacillus acidophilus, Enterococcus faecalis, Ent. faecium, Escherichia coli, Clostridium perfringens, Cl. butyricum, Cl. innocuum, Cl. bifermentans. Lactobacillus and Cl. bifermentans populations never rose above minimum detection limits (log10 2.0 and 4.0, respectively) under the experimental conditions employed in these studies. Inclusion of Des. desulfuricans in bacterial cultures (c. log10 8.4 viable cells ml-1) resulted in marked reductions (i.e. greater than 1 log) in planktonic cell population densities of several species, particularly Bif. longum, Cl. perfringens and Bif. pseudolongum. The two bacteroides species were unaffected by Des. desulfuricans, while numbers of Cl. butyricum increased. Extensive wall growth developed in the SRB culture, consisting mainly of Des. desulfuricans (log10 9.2 viable cells ml-1), Bact. thetaiotaomicron and Bact. vulgatus, with lesser numbers of facultative anaerobes, Cl. perfringens and Bif. longum. Wall growth was associated with a reduction in planktonic cell mass and increased acid production by the cultures. Chemotaxonomic study of chemostat microbiotas, on the basis of cellular fatty acid methyl ester (FAME) analyses, showed the existence of characteristic bacteroides (C15) and bifidobacterial (C18) markers, but desulfovibrio markers (i-C15:0, C16:0, i-C17:1) could be identified. The metabolic activities of saccharolytic organisms were altered in the SRB chemostat, including synthesis of a number of hydrolytic enzymes involved in carbohydrate breakdown, such as alpha-galactosidase, alpha-glucosidase and beta-galactosidase, together with several mucinolytic enzymes. High concentrations of sulphide (8.2 mmol 1-1) were detected in the SRB chemostat, suggesting that this metabolite may have been inhibitory to some species. Saccharolytic organisms growing in the SRB fermenter utilized more starch, but less galactose-containing polymers, which correlated with the observed glycosidase activities. Profound differences were also recorded with respect to fermentation product formation in the chemostats, where a major switch to acetate production occurred in the SRB culture, with concomitant reductions in propionate, butyrate and lactate, which is an important electron donor for desulfovibrios.     

Fractionation and analysis of rat liver14CH3-lecithins labeled in vivo

The¹⁴CH₃-lecithins were biosynthesized by normal adult rats injected with¹⁴CH₃-methionine. About 20% of the dose was incorporated into liver lecithins. The¹⁴CH₃-lecithins were isolated by thin-layer chromatography. Separation of lecithins on AgNO₃-treated silica gel yielded lecithins containing a saturated fatty acid in combination with mainly one unsaturated fatty acid, namely, oleic, linoleic, eicosatrienoic, or arachidonic acid. These fractions were eluted with methanolic choline chloride, which prevented elution of AgNO₃. The lecithins, after extraction into petroleum ether, were analyzed for radioactivity and for fatty acid composition. Yields were about 75%, based upon fatty acids or radioactivity applied to the plate. Specific activities differed sharply between the fractions, and arachidonoyllecithins had the highest specific activity. The sum of the activities contributed by each of the fractions agreed well with the specific activity of total lecithins, indicating the recovery of intact lecithin molecules. The recovery of intact molecules allows this procedure to be used with lecithins containing any isotopic labels. The high specific activity of arachidonoyl-lecithins relative to the other fractions indicates a high degree of specificity in the metabolic reactions which lead to the formation of rat liver lecithins.     

Inkjet printing of photocatalytically active TiO2 thin films from water based precursor solutions

In this work, aqueous chemical solution deposition route suited for inkjet printing is used for the synthesis of photocatalytically active TiO₂ coatings. Environmentally friendly precursor solutions with electromagnetic ink-jet printing, allows cheap and simple processing of TiO₂ films on glass. The hydrolysis reaction of water sensitive titanium alkoxide (Ti-alkoxide) precursor is controlled by adding complexing agents as citric acid and triethanolamine prior to water addition, and aqueous stable solutions are achieved. The pH of the solutions is brought to neutral to guarantee flexible processing, avoid damage to substrates and equipment. Solution parameters are adapted to obtain optimal gelation conditions and good jettability. The influence of processing parameters on the phase formation and surface morphology is studied by thermogravimetric analysis and differential thermal analysis (TGA/DTA), X-ray diffraction (XRD), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The photocatalytic activity of the films is evaluated by the degradation of methyl orange.     

Leaf Surface Compounds and Oviposition Preference of Turnip Root Fly Delia floralis: The Role of Glucosinolate and Nonglucosinolate Compounds

The role of leaf surface compounds influencing the oviposition of the turnip root fly, Delia floralis, was investigated using bioassays and fractionation of leaf surface extracts from four Brassica genotypes. Polar leaf surface extracts contained between 65 and 175 nM/g leaf equivalent of glucosinolates. However, following fractionation it was found that nonglucosinolates were the major stimuli for D. floralis oviposition. Electrophysiological studies of leaf surface extracts and their fractions were performed by using D. radicum, the cabbage root fly, as an analytical tool. The most behaviorally active fractions contained stimulatory compound(s) that had an activity profile identical to that previously described for recently discovered nonglucosinolate compounds. The role of leaf surface chemicals in influencing antixenotic resistance to D. floralis is discussed.     

Dialysis and gel filtration of isolated low density lipoproteins do not cause a significant loss of low density lipoprotein tocopherol and carotenoid concentration

The resistance of isolated low density lipoprotein (LDL) to copper-initiated oxidation is often used as a measure of effectiveness of an antioxidant intervention. Prior to oxidation excess salt and EDTA are removed via dialysis or gel filtration of the LDL sample. However, there is concern over whether the antioxidant content of dialyzed or gel-filtered LDL is truly representative of native LDL extracted from a blood sample. Previously, the experiments done after the storage of native and dialyzed LDL at −80°C showed that the dialysis step can cause a loss of up to 60% in the tocopherol and carotenoid content of LDL. In the present study, a comparison of the micronutrient concentration in freshly prepared dialyzed and native LDL from 35 subjects showed that after the correction for cholesterol, only lycopene (13%, P<0.001) and to a lesser extent α-carotene (8%, P<0.02) were significantly decreased, and the absolute fall in concentration was far smaller than previously reported. Other experiments done with smaller numbers of samples suggested that there were minimal micronutrient losses following gel filtration and that it was important to include 10 μmol/L EDTA in the dialysis and elution buffer; otherwise micronutrient losses did occur. In summary, immediate dialysis of freshly isolated LDL in the presence of 10 μmol/L. EDTA does not cause any major loss in the concentration of tocopherol and most carotenoids.     

合成二氧化氯的催化剂和方法及该催化剂的制备

使用一种失活速率降低的催化剂用亚氯酸钠的水溶液合成二氧化氮。催化剂最好为钯,钯与其它钯系金属的混合物（如Ｐｂ＋Ｐｔ）或者是钯与第１副族金属的混合物（Ｐｂ＋Ａｕ）,所用的载体为可用第１主族金属的硕酸盐如（Ｋ２ＣＯ３）、第２主族金属的碳酸盐（如ＣａＣＯ３）或者是可转化成ＭｇＯ的镁盐改性的物质,载体还可以是第１主族金属的碳酸盐（如Ｋ２ＣＯ３）或者是可转化成ＭｇＯ的镁盐改性的物质,载体还可以是第１主族金属