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W artykule przedstawiono elektroforetyczne rozdziały kolagenu mięśni bydlęcych z zastosowaniem PAGE-SDS. Dominował kolagen typu I. W wyniku hydrolizy pepsyną uzyskiwano zwiększenie rozpuszczalności kolagenu.
Enzymatic hydrolysis was conducted to recover the potentially addible high protein hydrolysate from cod backbones. The commercially available alcalase and trypsyne was used for hydrolysis at enzyme concentrations of 2.5; 5; 10; 15; 20; 30 and 40 mg/g backbones. The enzymatic deproteinization was conducted for 24 and 48 h. All procedures were performed at pH 7.0 and 4°C. The yield of enzymatic hydrolysis increases with the growth of trypsine and alcalase concentration up to 20 and 10 mg/g backbone, respectively. The increase of these values did not influence the yield of enzymatic deproteinization. After the treatment by trypsyne, maximum recovery of protein from cod backbone was 60%, while by alcalase it was 55%. The yield of enzymatic hydrolysis was the same after 24 and 48 h. Collagen losses did not exceed 0.2%.
The aim of the investigation was to establish the functional properties of muscle protein isolates from Baltic cod (Gadus morhua) spine which were also compared with a commercially available pork protein preparation. The muscle proteins were extracted in mild condition with 0.1M NaOH solution at 4°C and subsequently were precipitated at pH 4.5. The amino acid composition of protein isolates is similar to fresh muscle protein. At the basic pH values, the obtained preparation is almost three times more soluble than the commercially available pork protein. Cod proteins were in 25 and 90% solubilized at pH 8 and 12, respectively. At these pH values, after an increase of the ionic strength of the solution to 0.35, the protein solubility decreased about 10 and 15%, respectively. Spinal proteins at a pH range of 6-12 have a 16-time higher foaming capacity than that for pork proteins. The obtained proteins also have a two-times lower oil holding capacity and almost 5.5-times lower water holding capacity in comparison with the commercial preparation. The preparation of muscle proteins from Baltic cod had an ability to hold 0.26 g oil or 0.94 g water per g of protein. Its quite good functional properties encourage continuing research on the deodorization method. After deodorization this protein preparation could find an application in the food industry.
The aim of the study was to establish the optimum conditions for recovering non-collagen proteins from the backbones of cods by solvent extraction. The proteins were extracted in three different ways: twice in 24 h with 5% sodium chloride or 0.1 M sodium hydroxide solution, or firstly using 5% sodium chloride solution over 24 h, and then 0.1 M sodium hydroxide solution in 24 h. Different ratios of backbone to solution were tested, 1 : 2; 1 : 4; 1 : 6; 1 : 8; 1 : 10; 1 : 12. All procedures were performed at 4°C. 0.1 M solution of sodium hydroxide was more effective in extracting protein than 5% solution of sodium chloride. A 100% yield of non-collagen protein was recovered from fresh backbone by double 24 h extraction with sodium hydroxide solution, while this was 70%with sodium chloride solution. About 80% of the protein was soluble when extraction was conducted in the first stage with sodium chloride solution and then with sodium hydroxide solution. After 5 months of storing the backbone at -18°C, protein recovery decreased by about 40% for sodium chloride solution and about 20% for sodium hydroxide solution, and about 30% for mixed extraction. The extraction yield had no influence on the ratio of extracted material to solution. Collagen losses during extraction did not exceed 0,4%.
INTRODUCTION: Extracellular potentials, such as the Local Field Potentials (LFPs), are routinely measured in numerous electrophysiological experiments. LFP can carry valuable information about the electric properties of the tissue, however analysis of the recorded signal is usually a complex task. Apart from basic preparation, such as bandpass filtering and artifact removal, many other analytic methods have been proposed for the LFP study. Here we discuss methods for estimation of electric sources and sinks in brain tissue (Current Source Density, CSD) and methods to estimate connectivity in small networks, and their utility in analysis of cortical recordings in rats. AIM(S): Comparison of the effective connectivity and the structure of sinks and sources in cortical columns during whisker stimulations. METHOD(S): Analytic methods: kernel Current Source Density and Modular Connectivity Factorization (MCF) applied to LFP recordings and simulated data from cortical column. Experimental methods: Simultaneous multielectrode in vivo recordings from both hemispheres of the rats brain. RESULTS: Preliminary studies show different distribution of the current sources in contralateral to ipsilateral hemisphere during whisker stimulation in rats. Comparison of the hemispheres from deprived rats shows an extension of the whisker representation in the barrel cortex receptive field. CONCLUSIONS: KCSD method showssignificant differences in current sources localization in contralateral to ipsilateral hemisphere. Modular Connectivity Factorization method applied to LFP recordings from simulated data separates cortical column layers into interpretable modules. Physiological interpretation of the results needs further validation on the cortical column model. FINANCIAL SUPPORT: Instytut Biologii Doświadczalnej im. Marcelego Nenckiego Polska Akademia Nauk, Warsaw, Poland, Uniwersytet Warszawski, Warsaw, Poland
BACKGROUNS AND AIMS: The study develops methodology of photothrombotic stroke (PtS) induction in cats visual cortex for future studies of post-stroke visual recovery. Considering its well known organisation and similarity to the human, cat visual system is a good model for spontaneous and supported brain reorganisation after ischemia. METHODS: Photothrombosis was used as a model of focal ischemic cortical stroke. Standardisation of the methodology of cortical PtS was performed on four experimental cats with parallel local field potential (LFP) recordings in and around the stroke core – before, during and after the infarct. Intravenously injected Bengal Rose was locally irradiated by cold light via an optic bundle placed on the skull, thinned skull or directly on dura surface. Different light source parameters and irradiation time were tested. Postoperatively isolated brains were preserved and frozen, cut and stained. Final position of the electrodes was monitored on dried 50 μm slices. The cytochrome oxidase (CO) activity and Nissl staining were used to monitor the state of the tissue injury. RESULTS: The aimed unilateral stroke was performed in the dorsal zone of the left marginal gyrus over the visual cortex on a border of the cortical areas 17 and 18. CO visualizes the areas of lower mitochondrial activity in the illuminated tissue of the irradiated cortex in comparison to the contralateral intact homotopic areas. The spontaneous LFP dynamic decreased for at least three hours within the irradiated cortex but not in opposite hemisphere or surrounding tissue. CONCLUSIONS: The 25 minutes of the unilateral irradiation directly to the dura surface with the light temperature 2750K resulted in the most accurate lesion covering all the width of the marginal gyrus and partially the sulcus area not spreading on adjacent gyri or further blood vessels. The infarct reaches the white matter without its pronounced injury. Supported by:ERA-NET:REVIS&MNiSW0292/IP1/2013/72.
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