scholarly journals A METHOD FOR DETERMINING TOTAL PROTEIN OF ISOLATED CELLULAR ELEMENTS AND CORRESPONDING TRITIUM RADIOACTIVITY

1968 ◽  
Vol 38 (3) ◽  
pp. 562-573 ◽  
Author(s):  
Edward Koenig
Keyword(s):  
Total Protein ◽  
Trichloroacetic Acid ◽  
High Efficiency ◽  
Monochromatic Light ◽  
Cellular Protein ◽  
Total Density ◽  
Rabbit Retina ◽  
Protein Mass ◽  
Specific Absorbance ◽  
Rod Cell

A method is described for the microanalysis of protein, obtained from isolated tissue elements, in the range of 500 µµg-500 mµg. The method entails solubilization of cellular protein with phosphoric acid and heat after extraction of acid-soluble compounds, lipids, and RNA. A procedure for the extraction and recovery of cellular RNA by the use of 40% trichloroacetic acid is presented. The solubilized protein, in the form of a microdroplet, is photomicrographed with monochromatic light at 230 mµ. Total density in the microdroplet is determined from calibrated photographic plates by microdensitometry, and is converted to protein mass by using an experimentally determined average specific absorbance value. A solubilized protein labeled with tritium can be recovered after photomicrography, combusted, and reduced to generate tritiated gas for high-efficiency tritium radiometry. Total protein was analyzed in (a) nerve cells of three different sizes from Deiters' nucleus of the rabbit; and the whole rod cell and rod cell nucleus of the rabbit retina.

The effect of energy supply on the contribution of lean tissue to total body protein mass in pigs slaughtered at 100 kg

1997 ◽  
Vol 65 (3) ◽  
pp. 509-513 ◽  
Author(s):  
N. Quiniou ◽  
J. Noblet
Keyword(s):  
Protein Content ◽  
Total Protein ◽  
Energy Supply ◽  
The Body ◽  
Lean Tissue ◽  
Large White ◽  
Body Protein ◽  
Protein Mass ◽  
Body Tissues ◽  
Ad Libitum

AbstractThe effect of energy supply between 45 and 100 kg body weight (BW) on the contribution of lean tissue (muscle plus intermuscular adipose tissue) to total protein mass was studied in Large White castrated males (cLW), crossbred Piétrain × Large White castrated males (cPPx) and boars (bPPx). The pigs were allocated to four energy levels (0·70, 0·80, 0·90 or 1·00 ad libitum) and kept in metabolism cages in experiment 1 or given food ad libitum and kept in individual pens in experiment 2. Daily protein supplies were calculated to be non-limiting for growth and identical for all pigs in experiment 1. Temperature was 23°C in both experiments. The pigs were slaughtered at 100 kg BW and physically dissected; the body tissues were chemically analysed. Taking into account housing conditions, the food intake of pigs in experiment 2 corresponded to 1·20 of ad libitum intake of pigs in experiment 1; data of both experiments were combined. The energy supply and the type of pig influenced significantly the protein content of empty BW (eBW) (170 g/kg on average), of lean (184g/kg on average) and non-lean compartment (eBW minus lean, 152 g/kg on average), the proportion of total protein deposited in lean (604 g/kg of total protein, on average) but not the protein content in fat-free eBW (209 g/kg on average). The fat-free eBW can be predicted as 4·8 times the body protein mass.

scholarly journals Genetic rescue models refute nonautonomous rod cell death in retinitis pigmentosa

2017 ◽  
Vol 114 (20) ◽  
pp. 5259-5264 ◽  
Author(s):  
Susanne F. Koch ◽  
Jimmy K. Duong ◽  
Chun-Wei Hsu ◽  
Yi-Ting Tsai ◽  
Chyuan-Sheng Lin ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Retinitis Pigmentosa ◽  
High Efficiency ◽  
Genetic Rescue ◽  
Rod Photoreceptors ◽  
Rods And Cones ◽  
Promising Treatment ◽  
Low Efficiency ◽  
Rod Cell

Retinitis pigmentosa (RP) is an inherited neurodegenerative disease, in which the death of mutant rod photoreceptors leads secondarily to the non-cell autonomous death of cone photoreceptors. Gene therapy is a promising treatment strategy. Unfortunately, current methods of gene delivery treat only a fraction of diseased cells, yielding retinas that are a mosaic of treated and untreated rods, as well as cones. In this study, we created two RP mouse models to test whether dying, untreated rods negatively impact treated, rescued rods. In one model, treated and untreated rods were segregated. In the second model, treated and untreated rods were diffusely intermixed, and their ratio was controlled to achieve low-, medium-, or high-efficiency rescue. Analysis of these mosaic retinas demonstrated that rescued rods (and cones) survive, even when they are greatly outnumbered by dying photoreceptors. On the other hand, the rescued photoreceptors did exhibit long-term defects in their outer segments (OSs), which were less severe when more photoreceptors were treated. In summary, our study suggests that even low-efficiency gene therapy may achieve stable survival of rescued photoreceptors in RP patients, albeit with OS dysgenesis.

scholarly journals Mechanical and Structural Properties of a Lightweight Concrete with Different Types of Recycling Coarse Aggregate

2019 ◽  
Vol 26 (1) ◽  
pp. 33-40
Author(s):  
Muyasser M. Jomaa’h ◽  
Baraa Thaer Kamil ◽  
Omer S. Baghabra
Keyword(s):  
Mechanical Properties ◽  
Tensile Strength ◽  
Compressive Strength ◽  
High Efficiency ◽  
Coarse Aggregate ◽  
Splitting Tensile Strength ◽  
Modulus Of Rupture ◽  
Unit Weight ◽  
Dry Density ◽  
Total Density

The light of the world’s technological development in the construction field and the continuous need to apply of a high-efficiency building materials because old methods is no longer is used after the advent of the solutions that characterized by fast applications and maximum protection in addition to reducing costs and increase the sustainability of the establishment and its design age. The lightweights of various installations are an urgent need to decrease the dead loads. Therefore, this study is specie locally focus on replacing the normal coarse aggregate with lightweight coarse aggregate (claystone (bonza), rubber, thermostone and polystyrene) in various volumetric ratios of (25, 50 and 75) % in addition to a preparation reference mix. For the purpose identifying and studying the important specifications the new concrete which contributes to the self-load reduction of the concrete by reducing the total density of the mixture, were prepared models of cylinders and standard prisms, to evaluate the compressive strength and the splitting tensile strength respectively, Also the modulus of rupture and the unit weight, where carried out. The results tests indicated that a drop in the mechanical properties of the concrete with increasing the lightweight coarse aggregate , mechanical properties values : compressive strength , rupture modulus, splitting tensile strength and flexural strength were between (10.66-28.99) MPa (1.122-3.372) MPa, (3.606-6.83) MPa and (20.101-25.874)MPa compared with a reference mixes (38.44MPa), (3.969MPa), (10.476MPa) and (26.940)MPa respectively for mixes of (25, 50 and75)% with different light coarse aggregate , also the values of an oven dry density were between (1665.5-2287.58)kg/m3 compared with reference mixes (2426.41kg/m³). The best concrete mix was (M7, M10) of low density (1598.4 kg/m3) and (1580.4) kg /m3 and the compression strength within the permissible limits (15.47) MPa.

Importance of Protein Denaturation to Thermochemical Ablation of Liver Tumors

2011 ◽  
Author(s):  
Shoji Mori ◽  
Rajagopal Aravalli ◽  
Jeunghwan Choi ◽  
Erik Cressman ◽  
John Bischof
Keyword(s):  
Hepatocellular Carcinoma ◽  
Total Protein ◽  
Protein Denaturation ◽  
Liver Tumors ◽  
Cellular Protein ◽  
Human Hepatoma ◽  
Single Session ◽  
Local Ablation ◽  
Chemical Ablation ◽  
Protein Denaturant

Solid tumors in the liver such as hepatocellular carcinoma, often are not amenable to chemotherapy or surgical therapies. Local “ablation” methods including thermal and chemical ablation are therefore options for treatment. Hyperthermic ablation methods (RF, microwave, HIFU and laser) can potentially accomplish treatment in a single session but are considerably more costly than chemicals. It was therefore of interest to investigate if a known protein denaturant such as urea would destroy liver tumors. Further, we wished to assess whether this destruction occurs by protein denaturation mechanisms similar to hyperthermic destruction. Specifically, Lepock showed that hyperthermic cell destruction occurs for many cells when overall protein denaturation is greater than 5% (i.e. survival drops from 95% to 5% beyond this range). In this study we report on the cytotoxicity of urea on human hepatoma HuH-7. We then quantify the amount of cellular protein denaturation associated with cytotoxicity and show that protein denaturation in excess of 5% of total protein must occur in order for significant cell killing.

Neuropeptide Y stimulates hypertrophy of adult ventricular cardiomyocytes

1994 ◽  
Vol 266 (5) ◽  
pp. C1271-C1277 ◽  
Author(s):  
B. C. Millar ◽  
K. D. Schluter ◽  
X. J. Zhou ◽  
B. J. McDermott ◽  
H. M. Piper
Keyword(s):  
Protein Synthesis ◽  
Neuropeptide Y ◽  
Ventricular Myocytes ◽  
Cellular Protein ◽  
Trophic Effect ◽  
Prolonged Incubation ◽  
Protein Mass ◽  
Adult Rat ◽  
Culture Models ◽  
First Time

It was investigated whether neuropeptide Y (NPY) could exert a trophic effect on ventricular myocytes isolated from the adult rat heart. Two different culture models were used: day 1 and 7 cultures of cardiomyocytes. In day 1 and 7 cultures, NPY caused an increase in cellular protein mass. In day 1 cultures, NPY (10 nM) increased the protein-to-DNA ratio within 24 h by 10.1 +/- 2.8% (P < 0.01), but did not stimulate the incorporation of [14C]phenylalanine into cell proteins. The degradation of proteins was retarded in presence of NPY, revealed by pulse-chase experiments. In day 7 cultures, NPY (10 nM) increased the protein-to-DNA ratio within 24 h by 33.9 +/- 5.0% (P < 0.01), increased the RNA-to-DNA ratio by 19.2 +/- 6.4%, and stimulated the incorporation of [14C]phenylalanine by 45.5 +/- 4.5% (P < 0.01). As in day 1 cultures, protein degradation was retarded. The specific activities of cytosolic creatine kinase and lactate dehydrogenase were increased in presence of NPY. This study demonstrates for the first time that NPY is a trophic factor for cardiomyocytes. NPY can cause an increase in cellular mass of protein, i.e., hypertrophy, by two mechanisms: 1) reduction of degradation of protein, found in day 1 and 7 cultures, and 2) stimulation of protein synthesis, observed only in day 7 cultures. The responsiveness of protein synthesis to NPY stimulation is induced during prolonged incubation in culture.

The FRAME Alternatives Laboratory Database. 1. In Vitro Basal Cytotoxicity determined by the Kenacid Blue Total Protein Assay

2006 ◽  
Vol 34 (2) ◽  
pp. 151-175 ◽  
Author(s):  
Richard Clothier ◽  
Elke Gottschalg ◽  
Silvia Casati ◽  
Michael Balls
Keyword(s):  
Cell Line ◽  
Total Protein ◽  
Cell Number ◽  
Cellular Protein ◽  
Teratocarcinoma Cell ◽  
Wide Range ◽  
Significant Difference ◽  
Order Of Magnitude ◽  
Basal Cytotoxicity

A database of over 280 chemicals has been compiled by using a mouse 3T3-L1 fibroblast-like cell line in exponential growth, exposed to chemicals for 72 hours in a 96-well tissue culture plate format, and determining cell number via the Kenacid blue (KB) assay for total protein. Ranking the chemicals according to their basal cytotoxicity, expressed as the concentration (mM) that inhibits increase in total cellular protein over 72 hours by 50% (the ID50 value) shows a wide range of ID50 values, from 0.00003mM to 10,096mM. This information includes the results for MEIC chemicals 1–50, and we have now added basal cytotoxicity data for 23 of the next 25 MEIC chemicals. When the neutral red uptake (NRU) assay was performed with the same cell cultures, before the KB assay, very similar indications of basal cytotoxicity were obtained. Comparisons between the results with 3T3-L1 cells and with a human fibroblast-like cell line, BCL-D1 showed a significant difference in order of magnitude of the ID50 value for only 5 of 52 chemicals. However, there was a difference in ID50 value of more than one order of magnitude for 8 of 24 chemicals tested with an undifferentiated teratocarcinoma cell line, F9.

Effects of low and high relative molecular protein mass on four methods for total protein determination in urine

Pathology ◽  
1990 ◽  
Vol 22 (2) ◽  
pp. 89-92 ◽  
Author(s):  
Chew W. Lim ◽  
Wayne N. Chisnall ◽  
Yvonne M. Stokes ◽  
Phillip M. Debnam ◽  
Michael J. Crooke
Keyword(s):  
Total Protein ◽  
Protein Determination ◽  
Protein Mass

175. The precipitation of the proteins in milk. I. Casein. II. Total proteins. III. Globulin. IV. Albumin and Proteose-peptone

1938 ◽  
Vol 9 (1) ◽  
pp. 30-41 ◽  
Author(s):  
Samuel J. Rowland
Keyword(s):  
Acetic Acid ◽  
Total Protein ◽  
Trichloroacetic Acid ◽  
Sodium Acetate ◽  
Acetic Acid Solution ◽  
Acid Mixture ◽  
Sodium Acetate Solution ◽  
Acetate Solution ◽  
Maximum Precipitation ◽  
Proteose Peptone

Improved methods have been evolved for the separation of the total protein, casein, albumin, globulin, and proteose-peptone substances of milk.I. The use of acetic acid-sodium acetate solutions for the precipitation of casein is elucidated, and it is shown that the maximum precipitation of casein is rapidly effected from milk samples of varying casein content by the addition to 10 ml. of milk of about 80 ml. of water at 40° C. and 1·0 ml. of 10% acetic acid solution, followed, after 10 min., by 1·0 ml. of N sodium acetate solution.This maximum precipitation was found to be 1·0–1·4% greater than by Moir's method, and 2·4–3·8% greater than by the method of the Association of Oflicial Agricultural Chemists.A procedure is suggested for the determination of casein which avoids the tedious transfer and washing of the precipitate, and gives enhanced ease, accuracy and speed of working.II. The advantages of trichloroacetic acid for the precipitation of proteins in determinations of the total protein and the non-protein nitrogenous substances of milk are discussed.The trichloroacetic acid methods at present in use are shown not to give complete precipitation, and for this a rapid method employing, at room temperature, a final concentration of 12% acid in the milk-acid mixture is recommended.III. An accurate method for the precipitation of globulin uncontaminated with either albumin or casein is described.IV. Methods are given for the precipitation and separation of the albumin and proteose-peptone substances.

scholarly journals Distribution of proteins between nucleus and cytoplasm of amoeba proteus

1981 ◽  
Vol 88 (3) ◽  
pp. 516-525 ◽  
Author(s):  
L Goldstein ◽  
C Ko
Keyword(s):  
Amoeba Proteus ◽  
Cellular Protein ◽  
The Other ◽  
Nuclear Transplantation ◽  
Main Function ◽  
Functional Sites ◽  
Protein Mass ◽  
High Affinity ◽  
Nuclear Binding

By transplanting nuclei between labeled and unlabeled cells, we determined the localization of the major proteins of amebas and described certain features of their intracellular distributon. We identified approximately 130 cellular proteins by fluorography of one-dimensional polyacrylamide electrophoretic gels and found that slightly less than half of them (designated NP, for nuclear proteins) are almost exclusively nuclear. About 95 percent of the other proteins (designated CP for cytoplamsic proteins) are roughly equally concentrated in nucleus and cytoplasm, but - because the cytoplasm is 50 times larger than the nucleus - about 98 percent of each of the latter is in the cytoplasm. Of the CP, roughly 5 percent are not detectable in the nucleus. Assuming that these are restricted to the cytoplasm only because, for example, they are in structures too large to enter the nucleus and labeled CP readily exit a nucleus introduced into unlabeled cytoplasm, we conclude that the nuclear envelope does not limit the movement of any nonstructural cellular protein in either direction between the two compartments. Some NP are not found in the cytoplasm (although ostensibly synthesized there) presumably because of preferential binding within the nucleus. Almost one half of the protein mass in nuclei in vivo is CP and apparently only proteins of that group are lost from nuclei when cells are lysed. Thus, while an extracellular environment allows CP to exit isolated nuclei, the nuclear binding affinities for NP are retained. Further examination of NP distribution shows that many NP species are, in fact, detectable in the cytoplasm (although at only about 1/300 the nuclear concentration), apparently because the nuclear affinity is relatively low. These proteins are electrophoretically distinguishable from the high-affinity NP not found in the cytoplasm. New experiments show that an earlier suggestion that the nuclear transplantation operation causes an artifactual release of NP to the cytoplasm is largely incorrect. Moreover, we show that cytoplasmic "contamination" of nuclear preparations is not a factor in classifying proteins by these nuclear transplantation experiments. We speculate the no mechanism has evolved to confine most CP to the cytoplasm (where they presumably function exclusively) because the cytoplasm's large volume ensures that CP will be abundant there. Extending Bonner's idea of "quasi-functional nuclear binding sites" for NP, we suggest that a subset of NP usually have a low affinity for available intranuclear sites because their main function(s) occurs at other intranuclear sites to which they bind tightly only when particular metabolic conditions demand. The other NP (those completely absent from cytoplasm) presumable always are bound with high affinity at their primary functional sites.

scholarly journals Estimation of the kinetic parameters of whey proteins proteolysis in the UF-concentrate of cheese whey

2021 ◽  
Vol 82 (4) ◽  
pp. 107-112
Author(s):  
E. I. Melnikova ◽  
E. V. Bogdanova
Keyword(s):  
Total Protein ◽  
Cheese Whey ◽  
Whey Proteins ◽  
Experimental Studies ◽  
Skim Milk ◽  
Ceramic Membranes ◽  
Total Protein Content ◽  
Enzyme Preparations ◽  
Protein Mass ◽  
Wide Range

The purpose of the research is to substantiate the choice of enzyme preparations Promod 523MDP, Promod 439L, Flavorpro 766MDP, and Flavorpro 750MDP (Biocatalysts Limited, UK) and to determine the effective time of whey proteins hydrolysis in an ultrafiltration concentrate (UF-concentrate) of cheese whey for reducing their allergenicity based on the analysis of kinetic constants of the proteolysis reaction. Experimental studies were carried out with samples of cheese whey UF-concentrate with a total protein mass fraction at least 3.0% obtained with the use of industrial ultrafiltration unit MMS Swissflow UF with ceramic membranes under the conditions of the PSC Dairy Plant “Voronezhskii”. They were preliminarily subjected to enzymatic hydrolysis for 8 hours at a constant temperature, based on the dosage data, the optimum pH and the temperature of the used enzymes, recommended by the manufacturer. The specificity constant Vmax/Km was used to estimate the effectiveness of the enzyme preparations, which characterizes the constants of all stages of the hydrolysis reaction. The highest proteolysis rate has a mixture of Promod 439L and Flavorpro 766MDP in the ratio of 1.5 and 3.0%, respectively, of the total protein content in the substrate. Microscopy results showed an increasing in the solubility of nitrogen-containing components after hydrolysis due to a decreasing in hydrophobic areas on the surface of peptides. The resulting hydrolysate can be applied in the technology of a wide range of dairy products to reduce their residual antigenicity by partially replacing skim milk in the formulation.

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