Total protein content and protein synthesis within pre-elongation stage bovine embryos

Author(s):  
J.G. Thompson ◽  
A.N.M. Sherman ◽  
N.W. Allen ◽  
L.T. McGowan ◽  
H.R. Tervit
1995 ◽  
Vol 198 (5) ◽  
pp. 1071-1077 ◽  
Author(s):  
T Gomi ◽  
T Okuda ◽  
S Tanaka

The development and degeneration of the flight muscles in adult crickets, Gryllus bimaculatus, were studied (1) by determination of the total protein content, (2) by SDS one-dimensional polyacrylamide gel electrophoresis (SDS­PAGE) of muscle protein and (3) by in vitro culturing of the muscle. The total protein content of the dorso-longitudinal muscle (DLM) and metathoracic dorso-ventral muscle (DVM) increased during the early days of adult life in both sexes. This high protein content was maintained for at least a further 10 days in some individuals, while in others it declined to a low level. Mesothoracic DVMs in males also showed an increase in protein content after adult emergence but did not undergo histolysis, whereas those in females showed no significant temporal change in protein content. Removal of hind wings or artificial de-alation was found to be useful in inducing degeneration of DLMs and metathoracic DVMs. This treatment also stimulated ovarian development in females. An analysis by SDS­PAGE provided no evidence for new protein synthesis prior to or during flight muscle degeneration. A high rate of [3H]- or [35S]methionine incorporation was observed in DLMs taken from newly emerged adults, but, in intact crickets, the rate declined rapidly during the first 3 days of adult life, a pattern consistent with that obtained from the measurement of total protein content. Compared with DLMs removed from intact crickets, DLMs taken from de-alated crickets showed reduced rates of protein synthesis during in vitro culturing. This, together with the onset of protein degradation, appears to cause the rapid decrease in total protein content of the muscle in de-alated crickets.


2001 ◽  
Vol 101 (3) ◽  
pp. 295-304 ◽  
Author(s):  
Michael J. O'LEARY ◽  
Colin N. FERGUSON ◽  
Michael J. RENNIE ◽  
Charles J. HINDS ◽  
John H. COAKLEY ◽  
...  

We have investigated sequential changes in skeletal muscle and hepatic protein synthesis following sepsis, and their relationship to changes in circulating and tissue glutamine concentrations. Male Wistar rats underwent caecal ligation and puncture (CLP) or sham operation, with starvation, and were killed 24, 72 or 96 h later. A group of non-operated animals were killed at the time of surgery. Protein synthesis was determined using a flooding dose of l-[4-3H] phenylalanine, and glutamine concentrations were measured by an enzymic fluorimetric assay. Protein synthesis in gastrocnemius muscle fell in all groups. Gastrocnemius total protein content was reduced after CLP and at 72 and 96 h after sham operation. After CLP, protein synthesis was lower at 24 h, and total protein content was lower at 72 and 96 h, than in sham-operated animals. CLP was associated with increased liver protein synthesis at all time points, whereas there was no change after sham operation. Liver protein content did not change after CLP, but was lower at 72 and 96 h after sham operation than in non-operated animals. Plasma glutamine concentrations were reduced at 24 h after sham operation, and at 72 and 96 h after CLP. Muscle glutamine concentrations were reduced in all groups, with the decrease being greater following CLP than after sham operation. In the liver, glutamine concentrations were unchanged after CLP, but increased after sham operation. In rats with sepsis, decreases in muscle protein synthesis and content are associated with markedly reduced muscle glutamine concentrations. Plasma glutamine concentrations are initially maintained, but fall later. In liver, protein synthesis is increased, while glutamine concentrations are preserved. These results support a peripheral-to-splanchnic glutamine flux in sepsis.


Author(s):  
C H Knight

The aim of this review is to assess to what extent the biological processes of milk protein synthesis and secretion might be manipulated to the benefit of the milk producer, processor and consumer. There is considerable impetus for such an analysis. A completely new ‘agroceutical’ industry is in prospect, utilising novel biotechnology to produce high-value, non-mammary proteins in the milk of transgenic dairy animals (Carver et al., 1993). Nearer to home, the milk processing industry and the consumer may wish to see more modest modification of endogenous milk proteins, making use of naturally occurring genetic variants or, again, using biotechnological approaches (Dalgleish, 1992). Exciting as such possibilites are, I intend to set them to one side and concentrate on the issue that most immediately concerns the dairy farmer: how can he increase his milk protein concentration and, thereby, his profitability. Retrospective analysis reveals some small scope for improvement; between 1985 and 1990, compositional quality data (the basis for milk payment) revealed an increase in protein content from 32.6 to 32.8 g/1, an improvement of 0.6%. Over the same period, milk fat increased by 3.1% and recorded milk yields by 6.4% (all data from UK Dairy Facts and Figures, Federation of United Kingdom Milk Marketing Boards), suggesting that protein content is least amenable to manipulation. Gross production figures of this sort can be misleading, but more controlled experimentation supports the contention. By reducing the forage to concentrate ratio, Rook et al. (1992) increased total protein yield by an impressive 200 g/d. However, only one-quarter of this effect was due to improved protein content, the remainder coming from increased milk yield. Endocrine manipulation of milk production gives a similarly discouraging picture. Growth hormone treatment undoubtedly increases protein yield, but it does so by increasing the volume of milk produced with no detectable effect on protein content (Bauman, 1992). Milk production is also responsive to milking frequency but, once again, the effect is on yield of milk rather than on the protein content of that milk (Hillerton et al., 1990). So, where is the silver lining? Does biotechnology have the answer; can the presence, in mammary cells, of extra copies of correctly expressed milk protein or foreign protein genes markedly increase protein content? In Edinburgh, transgenic sheep are producing milk containing as much as 30 g/1 of human alpha- 1-antitrypsin (Carver et al.. 1993), representing some 50% or the total protein content (Wright et al., 1991). There is good anecdotal reason for believing that total protein content is elevated (one sheep reached 70 g/1), but the definitive statistical analysis is, so far, lacking. We have examined mammary function in transgenic mice expressing the sheep beta-lactoglobulin gene and compared them with non-transgenic controls (Wilde et al.,1992). Expression was good: beta-lactoglobulin accounted for 30% of all the protein present. But, at 95 and 106 mg/ml. respectively, transgenic mouse milk contained no more total protein than that of control mice. The encouraging feature of this work was that mammary tissue from transgenic mice cultured in vitro synthesised 41 % more total protein than did tissue from control mice, a significant difference (P ˂ 0.05).It would appear that the tissue itself had the intrinsic capability for increased protein synthesis, but a block or some sort was imposed in vivo. The next step is to discover the nature of that block, and to do so we must understand the complex series of events leading ultimately to protein secretion into the alveolar lumen.


2014 ◽  
Vol 50 (1-2) ◽  
pp. 33-38 ◽  
Author(s):  
Wiesława B. Chwirot ◽  
Alicja Górska-Brylass

The variations of the total protein content and protein synthesis were analyzed in successive stages of microsporogenesis in <em>Larix europaea</em> L. Pure fractions of microsporocytes containing the cells in successive phases of meiosis and fractions of the anther wall cells were used to perform the assays. It was revealed, that the protein content and the intensity of protein synthesis undergo considerable changes in the course of meiosis. It was proved also, by means of electrophoretic separation of protein fractions, that no new proteins are synthesised during differentiation of microsporocytes of larch, and that the microsporogenesis process is related rather to disappearance of some protein fractions.


2005 ◽  
Vol 33 (3) ◽  
pp. 207-213 ◽  
Author(s):  
Paul J. Dierickx

In our previously described Hep G2/24-hour/total protein assay, protein levels were measured by using the Lowry method. This assay was the best acute in vitro assay for the prediction of human toxicity within the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) study. In order to increase the MEIC data-base with a wider range of chemicals, we were interested in introducing the more practical 3-(4-carboxybenzoyl)-quinoline-2-carboxaldehyde (CBQCA) method for the quantification of the total protein content. Therefore, we investigated whether the same good results for the prediction of acute human toxicity would be obtained with the CBQCA method. The cells were treated for 24 hours, then cytotoxicity was determined by measuring the total protein content with CBQCA. The results were quantified by using the PI50c: the concentration (in mM) of test compound required to reduce the total protein content measured with the CBQCA-method by 50% as compared to the control cells. The results were compared with the PI50, the corresponding value when the Lowry method was used. A relatively low correlation was observed between PI50 and PI50c, reflecting the large and unexpected, differences when using the two protein assays. However, when comparing the log PI50c with the human toxicity, a correlation coefficient of r2 = 0.761 ( n = 44) was obtained for exactly the same series of MEIC chemicals. This value is clearly higher than that for the Lowry method ( r2 = 0.695). Compared to the Lowry method originally used, the Hep G2/24-hour/CBQCA total protein assay has the additional important advantage that it can be very easily adapted for large-scale analyses with robotic systems, including the on-line calculation of the results.


2002 ◽  
Vol 16 (3) ◽  
pp. 209-215 ◽  
Author(s):  
Adalberto Luiz Rosa ◽  
Márcio Mateus Beloti ◽  
Richard van Noort ◽  
Paul Vincent Hatton ◽  
Anne Jane Devlin

Hydroxyapatite (HA) has been used in orthopedic, dental, and maxillofacial surgery as a bone substitute. The aim of this investigation was to study the effect of surface topography produced by the presence of microporosity on cell response, evaluating: cell attachment, cell morphology, cell proliferation, total protein content, and alkaline phosphatase (ALP) activity. HA discs with different percentages of microporosity (< 5%, 15%, and 30%) were confected by means of the combination of uniaxial powder pressing and different sintering conditions. ROS17/2.8 cells were cultured on HA discs. For the evaluation of attachment, cells were cultured for two hours. Cell morphology was evaluated after seven days. After seven and fourteen days, cell proliferation, total protein content, and ALP activity were measured. Data were compared by means of ANOVA and Duncan’s multiple range test, when appropriate. Cell attachment (p = 0.11) and total protein content (p = 0.31) were not affected by surface topography. Proliferation after 7 and 14 days (p = 0.0007 and p = 0.003, respectively), and ALP activity (p = 0.0007) were both significantly decreased by the most irregular surface (HA30). These results suggest that initial cell events were not affected by surface topography, while surfaces with more regular topography, as those present in HA with 15% or less of microporosity, favored intermediary and final events such as cell proliferation and ALP activity.


1979 ◽  
Vol 236 (2) ◽  
pp. H268-H272 ◽  
Author(s):  
R. C. Hickson ◽  
G. T. Hammons ◽  
J. O. Holoszy

Adult female rats were exercised by daily swimming. All the increase in heart weight induced by the exercise occurred within 14 days and averaged 30%. The half times of the increases in heart weight and total protein content were about 4.5 days, whereas that of cytochrome c, which was used as a mitochondrial marker, was 6.5 days. The total amounts of DNA and of hydroxyproline in the heart, which were used to evaluate the degree of connective tissue hyperplasia, increased only slightly (8% and 10%, respectively). Other animals were subjected to the same swimming program for 21 days. Groups of rats were killed at various time intervals after stopping exercise. Heart weight, total protein content, and total cytochrome c content decreased rapidly initially, with 60% of the total regression of hypertrophy occurring during the first week. Thereafter, heart weight fell more gradually toward the sedentary control value. The hydroxyproline content of the heart, which was increased 10%, did not decrease during the regression of the hypertrophy.


Foods ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2431
Author(s):  
Nicola Landi ◽  
Sara Ragucci ◽  
Antimo Di Maro

Cow, sheep and goat raw milk raised in Ailano and Valle Agricola territories (‘Alto Casertano’, Italy) were characterized (raw proteins, free and total amino acids content) to assess milk quality. Raw milk with the highest total protein content is sheep milk followed by goat and cow milk from both localities. Total amino acid content in cow, goat and sheep raw milk is 4.58, 4.81 and 6.62 g per 100 g, respectively, in which the most abundant amino acid is glutamic acid (~20.36 g per 100 g of proteins). Vice versa, the free amino acids content characteristic profiles are different for each species. In particular, the most abundant free amino acid in cow, sheep and goat raw milk is glutamic acid (9.07 mg per 100 g), tyrosine (4.72 mg per 100 g) and glycine (4.54 mg per 100 g), respectively. In addition, goat raw milk is a source of taurine (14.92 mg per 100 g), retrieved in low amount in cow (1.38 mg per 100 g) and sheep (2.10 mg per 100 g) raw milk. Overall, raw milk from ‘Alto Casertano’ show a high total protein content and are a good source of essential amino acids.


Development ◽  
1973 ◽  
Vol 30 (3) ◽  
pp. 647-659
Author(s):  
Hiroshi Imoh ◽  
Tsutomu Minamidani

The present paper reports basic data on DNA content, protein content, and protein synthesis in Triturus pyrrhogaster embryos during development from cleavage to the hatching stage. Except for measurements of DNA and total protein contents, embryos were labeled with sodium carbonate-14C for 10 h and fractionated into embryonic cell components, i.e. cytoplasmic mass, yolk and pigment granules, and nuclei, in a discontinuous density gradient of sucrose. The protein content and the radioactivity incorporated into protein were measured in each fraction. Those fractions combining protein soluble in buffer at pH 8·3 and in 0·25 N-HCl were further studied with polyacrylamide gel electrophoresis. In the newt embryo, four stages of active DNA increase were observed when cultured at constant temperature; they were gastrula, neurula, late tail-bud, and before-hatching stages. Total protein per embryo decreased from 3 to 2 mg during the development studied. The content of cytoplasmic soluble protein per embryo was low and constant throughout development. Synthesis of the fraction was observed at the earliest stage of development studied though the rate was not high and specific activity of the soluble protein increased during development. Qualitative changes in the newly synthesized protein were observed. With the yolk fraction, synthesis of protein, other than from probable contamination with the cytoplasmic fraction, was not detected and a detailed description was omitted. Changes were observed at two stages of development in the synthesis of nuclear protein soluble in buffer at pH 8·3, the first at gastrulation and the second at late tail-bud stage. The change at gastrulation seemed to be the start of syntheses of the nuclear soluble proteins, while quantitative enhancement rather than qualitative change was noticed at late tail-bud stage. Most of the nuclear protein soluble in 0·25 N-HCI was histone. The histone content increased in accordance with increase in the DNA content and the rate of DNA accumulation was accompanied by proportionate incorporation of radioactivity into histone. Among histone fractions, unique behaviour of the very lysine-rich histone was observed. The availability of [14C]sodium carbonate in rough estimations of protein synthesis in embryos and significance of the data obtained have been discussed.


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