Flemming Jessen

Antioxidant activity of Cod (Gadus morhua) protein hydrolysates: in vitro assays and evaluation in 5% fish oil-in-water emulsion.

Cod protein hydrolysates were fractionated according to the molecular mass into three fractions of >5 kDa, 3-5 kDa and <3 kDa using an ultrafiltration membrane system. The antioxidative activity of the crude hydrolysates and the fractions were investigated, both in in vitro assays (DPPH radical scavenging activity, reducing power, Fe²⁺ chelating activity and inhibition of lipid oxidation in liposome model system), and in 5% fish oil-in-water emulsions. The <3 kDa fractions had very good radical scavenging activity, Fe²⁺ chelating activity and reducing power while the fraction 3-5 kDa resulted in higher protection against oxidation in the liposome model system. When tested in 5% oil-in-water emulsions, all the fractions, including the crude protein hydrolysate, were able to protect fish oil against oxidation in an iron induced oxidation system. However, none of the peptide fractions were effective in preventing tocopherol loss and showed no tocopherol sparing property. Volatile oxidation products showed an interaction between the aldehydes and the protein/peptides added in the emulsions, and this needs further investigation.

Species identification of smoked and gravad fish products by sodium dodecylsulphate polyacrylamide gel electrophoresis, urea isoelectric focusing and native isoelectric focusing: a collaborative study

Abstract A collaborative study on the use of sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE), urea-isoelectric focusing (urea-IEF) and native isoelectric focusing for the identification of species of smoked salmonids, gravad salmonids and smoked eels was carried out by eight laboratories. With SDS-PAGE, minor changes took place in the profiles of the processed salmonid species making it impossible or very difficult to identify closely related species. With urea-IEF, there were fewer changes in the profiles due to processing and the system generally had greater species-discriminating power for the processed salmonids than SDS-PAGE. The profiles of the eel species as obtained on SDS-PAGE or urea-IEF were not affected by smoking. Urea-IEF had greater species-discriminating power than SDS-PAGE for the eel species. Native IEF was useful in providing supplementary identification on species difficult to identify by SDS-PAGE or by urea-IEF in the case of cold smoked products.

Trunk posture and trapezius muscle load while working in standing, supported-standing, and sitting positions

A study of standing, supported-standing (“riding” on a rounded seat), and sitting postures was carried out on persons simulating assembly work in places with poor leg space. These postures and the upper trapezius muscle load were examined using statometric and electromyographic methods, respectively. While supported-standing or sitting, the lumbar spine moved toward kyphosis, even where there was no backward rotation of the pelvis. In adopting the position for anteriorly placed work, the upper arms were raised 30° forward or more; then, if a greater reach was necessary, the trunk was flexed as well. It is concluded that if leg space is poor, variation between supported-standing and standing should be encouraged, and an ordinary office chair avoided. Working level should be arranged so that it is lower than 5 cm above elbow level if no arm/wrist support is possible.

Comparison of office chairs with fixed forwards or backwards inclining or tiltable seats

SummaryThree adjustments of an office chair seat: — one inclining +10‡ (forwards), one inclining − 5‡ (backwards), and one being freely tiltable from −8‡ to +19.5‡ — were investigated using two groups of healthy female workers in a field (n=12), and a laboratory study (n=10), respectively. The seat adjustments were examined with regard to effects on foot swelling, lumbar muscular load, backrest pressure and subjective acceptability. Desk-work and typing were compared according to lumbar muscular activity, seat movements (tiltable seat), and backrest pressure. Foot swelling tended to increase with increasing seat height but was not influenced by the ability to tilt the seat or not. With the different seat adjustments lumbar muscular activity did not change systematically in spite of greater backrest pressure when the seat inclined backwards. The tiltable seat was preferred to the others. Typing was associated with a more constrained and tens posture than desk work, because movements, transferred to the tiltable seat, decreased and the muscular load increased. Backrest pressure was highest during typing. A tendency towards gradually increasing restlessness (i. e. seat movements) and increasing forward inclination of the tiltable seat with time was observed.

Purification of proteins of the Na/Cl cotransporter from membranes of Ehrlich ascites cells using a bumetanide-sepharose affinity column

SummaryBumetanide-binding proteins were isolated from membranes of Ehrlich ascites tumor cells by affinity chromatography. An affinity column was constructed with the active moiety of bumetanide as a ligand using 4′-azidobumetanide, a photoactive analogue which inhibits Na/Cl cotransport in Ehrlich cells with high specificity. Covalent binding of the 4′-azidobumetanide with Sepharose was promoted by photolysis. Membranes isolated from Ehrlich cells were solubilized withn-octylglucoside. Solubilized proteins retarded by the affinity column were readily eluted by bumetanide. In reducing gels the major proteins eluted by bumetanide were ∼76 kDa and 38–39 kDa. There were also two proteins of 32 to 35 kDa eluted in lesser amounts. No proteins retarded by the affinity column were eluted with extensive washing without bumetanide. Furthermore, bumetanide eluted no proteins from a „control” column lacking the specific ligand. Upon rechromatography with bumetanide in solution, bumetanide-eluted proteins were not retarded, but their purity was increased by the retardation of contaminating proteins. Bumetanide-binding protein purified in this manner were characterized further by electrophoresis in nonreducing, nondenaturing gels.

Species identification of cooked fish by urea isoelectric focusing and sodium dodecylsulfate polyacrylamide gel electrophoresis: a collaborative study

The suitability and reliability of urea IEF and SDS-PAGE for the identification of cooked fish flesh was tested by a collaborative study among nine laboratories. Urea IEF was performed with CleanGels as well as with ImmobilineGels, and ExcelGels were used for SDS-PAGE, enabling all three types of gels to be run in the same flat bed electrophoresis chamber. By strictly following optimised standard operation procedures (SOPs), five unknown cooked samples had to be identified with each technique using a set of 10 raw reference samples. With urea IEF, only one out of 35 identifications was incorrect, and with SDS-PAGE a similar result was obtained. It was concluded that methods, as now developed, are suitable for checking the species declaration of fishery products.

A standardized method of identification of raw and heat-processed fish by urea isoelectric focusing: A collaborative study

A urea‐isoelectric focusing (urea‐IEF) method of identifying fish species in processed fishery products was investigated as an interlaboratory collaborative study. The technique was optimized with respect to (i) protein extraction conditions, composition of the extraction solution (urea and SDS solutions), determination of protein concentrations of the fish extracts (five tested methods); (ii) nature of gel (with carrier ampholytes and Immobilines), conditions of rehydration of commercial dry gels, urea concentration; (iii) staining conditions, Coomassie blue and silver staining. The results of various experiments were compared to select the most appropriate methodology, with respect to the discrimination power of differentiating species with the minimal influence of heat processing, reproducibility, speed, and ease of application. The method recommended meets the requirements of food control and customs laboratories.

Identification of the anion exchange protein of ehrlich cells: A kinetic analysis of the inhibitory effects of 4,4′-diisothiocyano-2,2′-stilbene-disulfonic acid (DIDS) and labeling of membrane proteins with3H-DIDS

SummaryIn Ehrlich ascites tumor cells 4,4′-diisothiocyano-2,2′-stillbene-disulfonic acid (DIDS) inhibits the chloride exchange both reversibly and irreversibly. The reversible inhibition is practically instantaneous and of a competitive nature withK1 about 2 μm at zero chloride concentration. This is succeeded by a slow irreversible binding of DIDS to the transporter, with a chloride dependence suggesting binding to the same site as for reversible DIDS binding/inhibition. To identify the membrane protein involved in anion exchange, cells were labeled with3H-DIDS. Incubation of cells for 10 min with 25 μm DIDS at pH 8.2 leads to more than 95% inhibition of the DIDS-sensitive chloride exchange flux when the chloride concentration is low (15mm). This condition was used for the3H-DIDS-labeling experiments. After incubation the cells were disrupted, the membranes isolated and solubilized, and the proteins separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The distribution of the3H-activity in the gel showed only one major peak, which could be related to protein with a mol wt of about 30,000 Daltons. The number of transport sites was estimated at about 400,000 per cell, and from the DIDS-sensitive chloride flux under steady-state conditions we calculate a turnover number of 340 ions per sec per site.

Combination of statistical approaches for analysis of 2-DE data gives complementary results

Five methods for finding significant changes in proteome data have been used to analyze a two-dimensional gel electrophoresis data set. We used both univariate (ANOVA) and multivariate (Partial Least Squares with jackknife, Cross Model Validation, Power-PLS and CovProc) methods. The gels were taken from a time-series experiment exploring the changes in metabolic enzymes in bovine muscle at five time-points after slaughter. The data set consisted of 1377 protein spots, and for each analysis, the data set were preprocessed to fit the requirements of the chosen method. The generated results were one list from each analysis method of proteins found to be significantly changed according to the experimental design. Although the number of selected variables varied between the methods, we found that this was dependent on the specific aim of each method. CovProc and P-PLS focused more on getting the minimum necessary subset of proteins to explain properties of the samples. These methods ended up with less selected proteins. There was also a correlation between level of significance and frequency of selection for the selected proteins.

Wrist support during typing ― a controlled, electromyographic study

The effects of wrist support on the trapezius and brachial extensor muscle loads during touch-typing were investigated with 12 secretaries, who were suffering pain in one or more of these muscles. They typed for 15 min at each of four experimental situations. The load on the trapezius was significantly greater with wrist support than without, and more marked the higher the support was adjusted. The radial extensor muscle load did not vary significantly. In spite of the higher trapezius load, most of the secretaries preferred a wrist support. Typing performance and body movements, estimated from the movements transferred to a tiltable chair-seat, were both unaffected by wrist support. Whether or not a wrist support should be used for typing cannot be concluded from the present investigation. However, it can be concluded that for touch-typing the keys should be positioned low - probably as low above the thighs as possible.