Characterization of matrix metalloprotease activities induced in the sea urchin extraembryonic matrix, the hyaline layer

2001 ◽  
Vol 79 (4) ◽  
pp. 461-468 ◽  
Author(s):  
Christopher Sharpe ◽  
John J Robinson

Hyaline layers, freshly prepared from one-hour-old embryos, were devoid of gelatin-cleavage activity. However, upon storage at 4°C, gelatin-cleavage activities appeared; three species of apparent mol mass 94[Formula: see text]117-, 90-, and 45-kDa were seen. All three species required zinc for activity. Using gel-exclusion chromatography we separated the 94[Formula: see text]117-, and 90-kDa species from the 45-kDa activity. The two higher mol mass species were inhibited by ethylenebis (oxyethylenenitrilo) tetraacetic acid and the lost activity was restored by calcium. Reconstitution of activity occurred with an apparent dissociation constant (calcium) of 5 µM. The presence of millimolar concentrations of magnesium had a minimal inhibitory effect on activity. The thermal denaturation profile of the higher mol mass gelatin-cleavage activity was significantly different in the presence and absence of calcium. Stabilization of these activities against thermal denaturation at 60°C occurred with an apparent dissociation constant (calcium) of 0.6 mM. Magnesium had no significant effect on the thermal denaturation profile. Collectively, these results suggest at least two different modes of interaction between calcium and the higher mol mass gelatinases. These conclusions are discussed in the context of the high calcium and magnesium concentrations present in the sea water environment of the sea urchin embryo.Key words: sea urchin, embryo, matrix metalloprotease, calcium.


2016 ◽  
Vol 10 (1) ◽  
pp. 782-793 ◽  
Author(s):  
Ridho Bayuaji ◽  
M Sigit Darmawan ◽  
Boedi Wibowo ◽  
NA Husin ◽  
Srie Subekti

This paper presents a study of the flexural strength of geopolymer concrete beam using high calcium content fly ash (FA) in marine environment, without high heat curing. Two series of beam specimens were loaded to failure to study the effect of chloride environment on the flexural strength of geopolymer concrete beams. Series I specimens were subjected to sea environment, whereas series II were kept at room temperature. Tests performed on concrete cylinders show that the sea water has no effect on compressive and splitting tensile stress of high calcium content FA based geopolymer concrete. However, the ratio of splitting and compressive strength for both series was approximately 44%, almost double than that of normal concrete. In addition, the flexural test of concrete beams shows that the average cracking load for series I specimens was 275% higher than that of series II. However, the ultimate load, crack pattern and deflection characteristic for both series were very similar.



Author(s):  
Sima Ajdar qizi Askerova

Monitoring of sea water condition is one of major requirements for carrying out the reliable ecological control of water environment. Monitoring networks contain such elements as sea buoys, beacons, etc. and are designated for measuringvarious hydrophysical parameters, including salinity of sea water. Development of specialized network and a separate buoy system for measuring thesea water salinity at different depths makes it possible to determine major regularities of processes of pollution and self-recovery of the sea waters. The article describes the scientific and methodological basics for development of this specialized network and questions of its optimal construction. It is well-known that at a depth of 30-45 m of the Caspian Sea salinity decreases and then at a depth of 45-60 m salinity is fully recovered. The mentioned changes of salinity at the relatively upper layer of sea waters is of special interest for studying the effect of ocean-going processes on the climate forming in the Caspian area. In terms of informativeness of measurements of surface waters salinity, the most informative is a layer ata 30-60 m depth, where inversion and recovery of salinity take place. It is shown that in most informative subrange of measurements, i. e. at a depth of 30-60 m optimization of regime of measurements complex should be carried out in order to increase the effectiveness of held researches. It is shown that at a depth of 35-50 m choice of the optimum regime of measurements makes it possible to obtain the maximum amount of information.



Author(s):  
Keizo Negi ◽  
Keizo Negi ◽  
Takuya Ishikawa ◽  
Takuya Ishikawa ◽  
Kenichiro Iba ◽  
...  

Japan experienced serious water pollution during the period of high economic growth in 1960s. It was also the period that we had such damages to human health, fishery and living conditions due to red tide as much of chemicals, organic materials and the like flowing into the seas along the growing population and industries in the coastal areas. Notable in those days was the issues of environment conservation in the enclosed coastal seas where pollutants were prone to accumulate inside due to low level of water circulation, resulting in the issues including red tide and oxygen-deficient water mass. In responding to these issues, we implemented countermeasures like effluent control with the Water Pollution Control Law and improvement/expansion of sewage facilities. In the extensive enclosed coastal seas of Tokyo Bay, Ise Bay and the Seto Inland Sea, the three areas of high concentration of population, we implemented water quality total reduction in seven terms from 1979, reducing the total quantities of pollutant load of COD, TN and TP. Sea water quality hence has been on an improvement trend as a whole along the steady reduction of pollutants from the land. We however recognize that there are differences in improvement by sea area such as red tide and oxygen-deficient water mass continue to occur in some areas. Meanwhile, it has been pointed out that bio-diversity and bio-productivity should be secured through conservation/creation of tidal flats and seaweed beds in the view point of “Bountiful Sea” To work at these challenges, through the studies depending on the circumstances of the water environment in the enclosed coastal seas, we composed “The Policy of Desirable State of 8th TPLCS” in 2015. We have also added the sediment DO into the water quality standard related to the life-environmental items in view of the preservation of aquatic creatures in the enclosed water areas. Important from now on, along the Policy, is to proceed with necessary measures to improve water quality with good considerations of differences by area in the view point of “Beautiful and bountiful Sea”.



1983 ◽  
Vol 61 (1) ◽  
pp. 175-189
Author(s):  
R. Kuriyama ◽  
G.G. Borisy

Conditions that induce the formation of asters in unfertilized sea-urchin eggs have been investigated. Monasters were formed by treatment of eggs with acidic or basic sea-water, or procaine- or thymol-containing sea-water. A second treatment step, incubation with D2O-containing, ethanol-containing or hypertonic sea-water induced multiple cytasters. The number and size of cytasters varied according to the concentration of agents and duration of the first and second treatments, and also upon the species of eggs and the season in which the eggs were obtained. Generally, a longer second treatment or a higher concentration of the second medium resulted in a higher number of cytasters per egg. Asters were isolated and then examined by light and electron microscopy. Isolated monasters apparently lacked centrioles, whereas cytasters obtained from eggs undergoing the two-step treatment contained one or more centrioles. Up to eight centrioles were seen in a single aster; the centrioles appeared to have been produced during the second incubation. Centrospheres prepared from isolated asters retained the capacity to nucleate the formation of microtubules in vitro as assayed by light and electron microscopy. Many microtubules radiated from the centre of isolated asters, whether they contained centrioles or not. This observation is consistent with many other reports that microtubule-organizing centres need not contain centrioles.



1997 ◽  
Vol 200 (22) ◽  
pp. 2881-2892 ◽  
Author(s):  
P Leong ◽  
D Manahan

Early stages of animal development have high mass-specific rates of metabolism. The biochemical processes that establish metabolic rate and how these processes change during development are not understood. In this study, changes in Na+/K+-ATPase activity (the sodium pump) and rate of oxygen consumption were measured during embryonic and early larval development for two species of sea urchin, Strongylocentrotus purpuratus and Lytechinus pictus. Total (in vitro) Na+/K+-ATPase activity increased during development and could potentially account for up to 77 % of larval oxygen consumption in Strongylocentrotus purpuratus (pluteus stage) and 80 % in Lytechinus pictus (prism stage). The critical issue was addressed of what percentage of total enzyme activity is physiologically active in living embryos and larvae and thus what percentage of metabolism is established by the activity of the sodium pump during development. Early developmental stages of sea urchins are ideal for understanding the in vivo metabolic importance of Na+/K+-ATPase because of their small size and high permeability to radioactive tracers (86Rb+) added to sea water. A comparison of total and in vivo Na+/K+-ATPase activities revealed that approximately half of the total activity was utilized in vivo. The remainder represented a functionally active reserve that was subject to regulation, as verified by stimulation of in vivo Na+/K+-ATPase activity in the presence of the ionophore monensin. In the presence of monensin, in vivo Na+/K+-ATPase activities in embryos of S. purpuratus increased to 94 % of the maximum enzyme activity measured in vitro. Stimulation of in vivo Na+/K+-ATPase activity was also observed in the presence of dissolved alanine, presumably due to the requirement to remove the additional intracellular Na+ that was cotransported with alanine from sea water. The metabolic cost of maintaining the ionic balance was found to be high, with this process alone accounting for 40 % of the metabolic rate of sea urchin larvae (based on the measured fraction of total Na+/K+-ATPase that is physiologically active in larvae of S. purpuratus). Ontogenetic changes in pump activity and environmentally induced regulation of reserve Na+/K+-ATPase activity are important factors that determine a major proportion of the metabolic costs of sea urchin development.



Development ◽  
1987 ◽  
Vol 101 (2) ◽  
pp. 255-265 ◽  
Author(s):  
J.A. Anstrom ◽  
J.E. Chin ◽  
D.S. Leaf ◽  
A.L. Parks ◽  
R.A. Raff

In this report, we use a monoclonal antibody (B2C2) and antibodies against a fusion protein (Leaf et al. 1987) to characterize msp130, a cell surface protein specific to the primary mesenchyme cells of the sea urchin embryo. This protein first appears on the surface of these cells upon ingression into the blastocoel. Immunoelectronmicroscopy shows that msp130 is present in the trans side of the Golgi apparatus and on the extracellular surface of primary mesenchyme cells. Four precursor proteins to msp130 are identified and we show that B2C2 recognizes only the mature form of msp130. We demonstrate that msp130 contains N-linked carbohydrate groups and that the B2C2 epitope is sensitive to endoglycosidase F digestion. Evidence that msp130 is apparently a sulphated glycoprotein is presented. The recognition of the B2C2 epitope of msp130 is disrupted when embryos are cultured in sulphate-free sea water. In addition, two-dimensional immunoblots show that msp130 is an acidic protein that becomes substantially less acidic in the absence of sulphate. We also show that two other independently derived monoclonal antibodies, IG8 (McClay et al. 1983; McClay, Matranga & Wessel, 1985) and 1223 (Carson et al. 1985), recognize msp130, and suggest this protein to be a major cell surface antigen of primary mesenchyme cells.



2002 ◽  
Vol 6 (5) ◽  
pp. 797-817 ◽  
Author(s):  
C. Neal ◽  
P. Shand

Abstract. A survey of surface, spring and borehole waters associated with the ophiolite rocks of Cyprus shows five broad water types (1) Mg-HCO3, (2) Na-SO4-Cl-HCO3, (3) Na-Ca-Cl-SO4-OH-CO3, (4) Na-Cl-SO4 and (5) Ca-SO4. The waters represent a progression in chemical reactivity from surface waters that evolve within a groundwater setting due to hydrolysis of the basic/ultrabasic rock as modified by CO2-weathering. An increase in salinity is also observed which is due to mixing with a saline end-member (modified sea-water) and dissolution of gypsum/anhydrite. In some cases, the waters have pH values greater than 11. Such high values are associated with low temperature serpentinisation reactions. The system is a net sink for CO2. This feature is related not only to the hydrolysis of the primary minerals in the rock, but also to CaCO3 or Ca-Mg-CO3 solubility controls. Under hyperalkaline conditions, virtually all the carbon dioxide is lost from the water due to the sufficiently high calcium levels and carbonate buffering is then insignificant. Calcium sulphate solubility controls may also be operative when calcium and sulphate concentrations are particularly high. Keywords: Cyprus, Troodos, ophiolite, serpentinisation, spring, stream, water quality, bromide, iodine, boron, trace elements, hyperalkaline.



1989 ◽  
Vol 257 (1) ◽  
pp. C29-C35 ◽  
Author(s):  
J. P. Manfredi ◽  
R. Marquetant ◽  
A. D. Magid ◽  
E. W. Holmes

The muscle isozyme of adenylosuccinate synthetase (AdSS), an enzyme of the purine nucleotide cycle, has previously been shown to bind to purified F-actin in buffers of low ionic strength and pH (Ogawa et al. Eur. J. Biochem. 85: 331-338, 1978). We have extended these observations by measuring the association of both crude and purified AdSS with the contractile proteins of muscle in buffers of physiological ionic strength and pH. Under these conditions, the enzyme binds to F-actin, actin-tropomyosin complexes, reconstructed thin filaments, and myofibrils but not to myosin. The apparent dissociation constant of 1.2 microM and binding maximum of 2.6 nmol enzyme/mg myofibrils indicate that binding of AdSS to myofibrils can be physiologically significant. The results suggest that AdSS in muscle may be associated with the thin filament of myofibrils.



1946 ◽  
Vol 29 (5) ◽  
pp. 267-275 ◽  
Author(s):  
Herbert Shapiro

A study was made of the diffusion of the red pigment echinochrome from the eggs of the sea urchin, Arbacia punctulata, into sea water. Unfertilized eggs retained their pigment, over periods of hours. Outward diffusion of pigment from unfertilized eggs normally is entirely negligible, or does not occur at all. Enchancing the calcium or potassium content of the artificial sea water (while retaining isosmotic conditions) did not induce pigment release. Under anaerobic conditions, unfertilized eggs release pigment in small quantities. Fertilization alone brings about echinochrome release. Fertilized eggs invariably released pigment, whether in normal sea water, or sea water with increased calcium or potassium. This diffusion of the pigment began during the first cleavage, possibly soon after fertilization. The pigment release is not a consequence solely of the cell's permeability to echinochrome (or chromoprotein, or other pigment combination) but is preceded by events leading to a release of echinochrome from the granules in which it is concentrated within the cell. These events may be initiated by activation or by anaerobiosis. The phenomenon was not due to cytolysis.



1951 ◽  
Vol 34 (3) ◽  
pp. 285-293 ◽  
Author(s):  
Anna Monroy Oddo ◽  
Maria Esposito

In the eggs of Arbacia lixula and Paracentrotus lividus an uptake of K occurs during the first 10 minutes following fertilization. Between 10 and 40 minutes K is then released. Both in Arbacia and in Paracentrotus the minimum point of the curve coincides with the nuclear streak stage. A maximum loss of 25 per cent in Arbacia and 20 per cent in Paracentrotus with respect to the amount present in the unfertilized eggs has been found. From 40 minutes up to 1 hour K undergoes a further increase and when the first cleavage sets in the same amount of K is present as in the unfertilized eggs. By treating the eggs with K-free artificial sea water it has been established that about 60 per cent of the K content of the eggs is in a non-diffusible condition. Also under such conditions the eggs when fertilized are able to take up even the very small amount of K present in the medium that was released by them prior to fertilization.



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