Biochemical Properties of Anionic Trypsin Acting at High Concentration of NaCl Purified from the Intestine of a Carnivorous Fish: Smooth Hound (Mustelus mustelus)

2010 ◽  
Vol 58 (9) ◽  
pp. 5763-5769 ◽  
Author(s):  
Ali Bougatef ◽  
Rafik Balti ◽  
Rim Nasri ◽  
Kemel Jellouli ◽  
Nabil Souissi ◽  
...  
1995 ◽  
Vol 129 (1) ◽  
pp. 219-231 ◽  
Author(s):  
T J Chilcote ◽  
T Galli ◽  
O Mundigl ◽  
L Edelmann ◽  
P S McPherson ◽  
...  

There is strong evidence to indicate that proteins of the synaptobrevin family play a key role in exocytosis. Synaptobrevin 1 and 2 are expressed at high concentration in brain where they are localized on synaptic vesicles. Cellubrevin, a very similar protein, has a widespread tissue distribution and in fibroblasts is localized on endosome-derived, transferin receptor-positive vesicles. Since brain cellubrevin is not detectable in synaptic vesicles, we investigated whether cellubrevin and the synaptobrevins are differentially targeted when co-expressed in the same cell. We report that in the nervous system cellubrevin is expressed at significant levels only by glia and vascular cells. However, cellubrevin is coexpressed with the two synaptobrevins in PC12 cells, a neuroendocrine cell line which contains synaptic vesicle-like microvesicles. In PC12 cells, cellubrevin has a distribution very similar to that of synaptobrevin 1 and 2. The three proteins are targeted to neurites which exclude the transferrin receptor and are enriched in synaptic-like microvesicles and dense-core granules. They are recovered in the synaptic-like microvesicle peak of glycerol velocity gradients, have a similar distribution in isopycnic fractionation and are coprecipitated by anti-synaptobrevin 2 immunobeads. Finally, cellubrevin, like the synaptobrevins, interact with the neuronal t-SNAREs syntaxin 1 and SNAP-25. These results suggest that cellubrevin and the synaptobrevins have similar function and do not play a specialized role in constitutive and regulated exocytosis, respectively.


1992 ◽  
Vol 285 (1) ◽  
pp. 85-90 ◽  
Author(s):  
C Job ◽  
D Shire ◽  
V Sure ◽  
D Job

Wheat-germ RNA polymerase II is able to catalyse a DNA-dependent reaction of RNA synthesis in the presence of a high concentration (1 mg/ml) of the fungal toxin alpha-amanitin. This anomalous reaction is specifically directed by single-stranded or double-stranded homopolymer templates, such as poly(dC) or poly(dC).poly(dG), and occurs in the presence of either Mn2+ or Mg2+ as the bivalent metal cofactor. In contrast, the transcription of other synthetic templates, such as poly(dT), poly(dA).poly(dT) or poly[d(A-T)] is completely abolished in the presence of 1 microgram of alpha-amanitin/ml, in agreement with well-established biochemical properties of class II RNA polymerases. Size analysis of reaction products resulting from transcription of (dC)n templates of defined lengths suggests that polymerization of RNA chains proceeds through a slippage mechanism. The fact that alpha-amanitin does not impede this synthetic reaction implies that the amatoxin interferes with the translocation of wheat-germ RNA polymerase II along the DNA template.


Agronomy ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 409
Author(s):  
Murtaza Khan ◽  
Nkulu Kabange Rolly ◽  
Tiba Nazar Ibrahim Al Azzawi ◽  
Muhammad Imran ◽  
Bong-Gyu Mun ◽  
...  

Lead (Pb) is one of the major environmental heavy metal pollutants, known as being neither essential nor beneficial for any living organisms, and which is detrimental to plant fitness, growth, and productivity, as well as human health. This study investigated the changes in the morphological, physiological, and biochemical properties of rice cultivars exposed to lead (Pb). Therefore, soil was contaminated with a solution containing 0.6 mM or 1.2 mM Pb four weeks prior to transplanting. Then, 4-week-old rice seedlings of Tunnae, Ilmi, Yasmen, Mashkab, and Amber Barka were transplanted into the contaminated soil and grown until maturity. The results showed that a high concentration of lead (1.2 mM) induced significant reduction in the plant height, number of tillers, number of panicles per plant, and the number of spikelets per panicle in Pb-sensitive rice cultivars, while in Pb-tolerant cultivars, a balanced growth of plants and non-significant change in the major yield components were recorded. However, all rice cultivars showed a reduced biomass dry weight. Under the same conditions, we observed a differential enzymatic antioxidant activity, with catalase (CAT) and peroxidase (POD) being the most active. In addition, the proline accumulation and sucrose content increased concomitant with an increase in the Pb concentration, while the total protein and chlorophyll contents significantly decreased. Of all the soluble sugars analyzed, sucrose was the most abundant in response to Pb treatment. Interestingly, the rice cultivars Tunnae and Mashkab exhibited a high degree of tolerance towards Pb stress, with a balanced plant height, number of tillers, number of panicles, and number of spikelets per plant. Therefore, all results collectively suggest that the tolerance to Pb-induced oxidative stress observed in Tunnae and Mashkab could be a result of a synergetic action of both enzymatic and non-enzymatic antioxidant systems, leading to a balanced reduction–oxidation status in rice.


PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8771
Author(s):  
Rebecca Sternke-Hoffmann ◽  
Amelie Boquoi ◽  
David Lopez Y. Niedenhoff ◽  
Florian Platten ◽  
Roland Fenk ◽  
...  

In light chain (LC) diseases, monoclonal immunoglobulin LCs are abundantly produced with the consequence in some cases to form deposits of a fibrillar or amorphous nature affecting various organs, such as heart and kidney. The factors that determine the solubility of any given LC in vivo are still not well understood. We hypothesize that some of the biochemical properties of the LCs that have been shown to correlate with amyloid fibril formation in patients also can be used as predictors for the degree of kidney damage in a patient group that is only biased by protein availability. We performed detailed biochemical and biophysical investigations of light chains extracted and purified from the urine of a group of 20 patients with light chain disease. For all samples that contained a sufficiently high concentration of LC, we quantified the unfolding temperature of the LCs, the monomer-dimer distribution, the digestibility by trypsin and the formation of amyloid fibrils under various conditions of pH and reducing agent. We correlated the results of our biophysical and biochemical experiments with the degree of kidney damage in the patient group and found that most of these parameters do not correlate with kidney damage as defined by clinical parameters. However, the patients with the greatest impairment of kidney function have light chains which display very poor digestibility by trypsin. Most of the LC properties reported before to be predictors of amyloid formation cannot be used to assess the degree of kidney damage. Our finding that poor trypsin digestibility correlates with kidney damage warrants further investigation in order to probe a putative mechanistic link between these factors.


2017 ◽  
Vol 5 (1) ◽  
pp. 9-16
Author(s):  
Atefeh Mohammadi ◽  
Mohammad Hussein Abnosi ◽  
Reza Pakyari

Background: Sodium nitroprusside (SNP) releases nitric oxide which has signaling role. Objectives: This study was conducted to understand the role of low concentration of SNP on viability, proliferation and biochemical properties of rat bone marrow mesenchymal stem cells (MSCs). Materials and Methods: MSCs were used to evaluate the viability and morphology in presence of SNP (1 to 100 µM) at 12, 24 and 36 hours. Then 10, 50 and 100 µM of SNP as well as 24 hours were selected for further study. Cell proliferation was investigated by colony forming assay and population doubling number (PDN). Calcium (Ca2+) potassium (K+) and sodium (Na+) level as well as activity of alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) were measured. Results: The MSCs viability increased when treatment with 1 and 10 µM at all the treatment periods while 90 and 100 µM caused significant reduction after 24 and 36 hours. Also 10 µM caused elevation whereas 50 and 100 µM showed reduction of proliferation ability. We observed morphological changes and significant reduction of all the investigated enzymes with 100 µM. Activity of ALT and AST were elevated with 10 µM after 24 hours, whereas LDH and ALP activities were not changed. Na+, K+ and Ca2+ was not changed due to 10 and 50 µM treatments, whereas 100 µM only elevated the level of calcium and sodium ions. Conclusions: Low concentration of SNP caused increase of viability and proliferation due to metabolic activity elevation. But the high concentration of SNP induced cell viability and proliferation reduction caused by metabolic and ionic imbalance as well as infrastructure alteration.


Author(s):  
Daniel C. Pease

A previous study demonstrated that tissue could be successfully infiltrated with 50% glutaraldehyde, and then subsequently polymerized with urea to create an embedment which retained cytomembrane lipids in sectioned material. As a result, the 180-190 Å periodicity characteristic of fresh, mammalian myelin was preserved in sections, as was a brilliant birefringence, and the capacity to bind OsO4 vapor in the hydrophobic bilayers. An associated (unpublished) study, carried out in co-operation with Drs. C.K. Akers and D.F. Parsons, demonstrated that the high concentration of glutaraldehyde (and urea) did not significantly alter the X-ray diffraction pattern of aldehyde-fixed, myelin. Thus, by itself, 50% glutaraldehyde has little effect upon cytomembrane systems and can be used with confidence for the first stages of dehydration.


Author(s):  
Glenn M. Cohen ◽  
Radharaman Ray

Retinal,cell aggregates develop in culture in a pattern similar to the in ovo retina, forming neurites first and then synapses. In the present study, we continuously exposed chick retinal cell aggregates to a high concentration (1 mM) of carbamylcholine (carbachol), an acetylcholine (ACh) analog that resists hydrolysis by acetylcholinesterase (AChE). This situation is similar to organophosphorus anticholinesterase poisoning in which the ACh level is elevated at synaptic junctions due to inhibition of AChE, Our objective was to determine whether continuous carbachol exposure either damaged cholino- ceptive neurites, cell bodies, and synaptic elements of the aggregates or influenced (hastened or retarded) their development.The retinal tissue was isolated aseptically from 11 day embryonic White Leghorn chicks and then enzymatically (trypsin) and mechanically (trituration) dissociated into single cells. After washing the cells by repeated suspension and low (about 200 x G) centrifugation twice, aggregate cell cultures (about l0 cells/culture) were initiated in 1.5 ml medium (BME, GIBCO) in 35 mm sterile culture dishes and maintained as experimental (containing 10-3 M carbachol) and control specimens.


Author(s):  
James Cronshaw

Long distance transport in plants takes place in phloem tissue which has characteristic cells, the sieve elements. At maturity these cells have sieve areas in their end walls with specialized perforations. They are associated with companion cells, parenchyma cells, and in some species, with transfer cells. The protoplast of the functioning sieve element contains a high concentration of sugar, and consequently a high hydrostatic pressure, which makes it extremely difficult to fix mature sieve elements for electron microscopical observation without the formation of surge artifacts. Despite many structural studies which have attempted to prevent surge artifacts, several features of mature sieve elements, such as the distribution of P-protein and the nature of the contents of the sieve area pores, remain controversial.


Author(s):  
R.A. Herring

Rapid thermal annealing (RTA) of ion-implanted Si is important for device fabrication. The defect structures of 2.5, 4.0, and 6.0 MeV As-implanted silicon irradiated to fluences of 2E14, 4E14, and 6E14, respectively, have been analyzed by electron diffraction both before and after RTA at 1100°C for 10 seconds. At such high fluences and energies the implanted As ions change the Si from crystalline to amorphous. Three distinct amorphous regions emerge due to the three implantation energies used (Fig. 1). The amorphous regions are separated from each other by crystalline Si (marked L1, L2, and L3 in Fig. 1) which contains a high concentration of small defect clusters. The small defect clusters were similar to what had been determined earlier as being amorphous zones since their contrast was principally of the structure-factor type that arises due to the difference in extinction distance between the matrix and damage regions.


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