scholarly journals Mucin biosynthesis. Properties of a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase

1985 ◽  
Vol 227 (2) ◽  
pp. 405-412 ◽  
Author(s):  
P W Cheng ◽  
W E Wingert ◽  
M R Little ◽  
R Wei
Keyword(s):  
Enzyme Activity ◽  
Freezing Point ◽  
Cetylpyridinium Chloride ◽  
Sodium Dodecyl ◽  
Sodium Deoxycholate ◽  
Gas Chromatography Mass Spectrometry ◽  
Tween 20 ◽  
Group A ◽  
Michaelis Constants ◽  
Triton X

We have characterized a bovine tracheal mucin beta-6-N-acetylglucosaminyltransferase that catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the C-6 of the N-acetylgalactosamine residue of galactosyl-β 1→3-N-acetylgalactosamine. Optimal enzyme activity was obtained between pH 7.5-8.5, at 5mM-MnCl2, and at 0.06-0.08% (v/v) Triton X-100 (or Nonidet P-40), or 0.5-5.0% (v/v) Tween 20. Ba2+, Mg2+ and Ca2+ could partially replace Mn2+, but Co2+, Fe2+, Cd2+ and Zn2+ could not. Sodium dodecyl sulphate, cetylpyridinium chloride, sodium deoxycholate, octyl beta-D-glucoside, digitonin and alkyl alcohols were less effective in enhancing enzyme activity, and dimethyl sulphoxide was ineffective. The apparent Michaelis constants were 1.25 mM for UDP-N-acetylglucosamine, 0.94-3.34 mM for freezing-point-depressing glycoprotein and 0.19 mM for periodate-treated blood-group-A porcine submaxillary mucin. Asialo ovine submaxillary mucin could not serve as the glycosyl acceptor. The structure of the 14C-labelled oligosaccharide obtained by alkaline-borohydride treatment of the product was identified as Gal beta 1→3(Glc-NAc beta 1→6)N-acetylgalactosaminitol by beta-hexosaminidase treatment, gas chromatography-mass spectrometry and 1H-n.m.r. (270 MHz) analysis. The enzyme is important in the regulation of mucin oligosaccharide biosynthesis.

scholarly journals Essential charged amino acids in the binding of fibronectin to gelatin

1982 ◽  
Vol 201 (1) ◽  
pp. 1-8 ◽  
Author(s):  
M Vuento ◽  
E Salonen ◽  
K Osterlund ◽  
U H Stenman
Keyword(s):  
Hydrophobic Interactions ◽  
Sodium Dodecyl ◽  
Sodium Deoxycholate ◽  
Binding Activity ◽  
Tween 20 ◽  
Ph Optimum ◽  
Ionic Detergents ◽  
Charged Amino Acids ◽  
Triton X

The binding of fibronectin to gelatin-agarose was strictly dependent on pH, having a pH optimum of 7-9. The binding was strongly inhibited by increasing ionic strength. A chemical modification of lysyl and arginyl groups of fibronectin abolished the binding activity. The anionic detergents sodium dodecyl sulphate and sodium deoxycholate in concentrations of 10-100mM had the same effect. The binding was not affected by the non-ionic detergents Triton X-100, Tween 20 or Lubrol WX. The results demonstrate an important role of ionic interactions in the binding of fibronectin to gelatin. Absence of inhibition by non-ionic detergents suggests that hydrophobic interactions contribute relatively little to the binding of fibronectin to gelatin.

scholarly journals The effect of chemical agents on the turnover of the bound phosphate associated with the sodium-and-potassium ion-stimulated adenosine triphosphatase in ox brain microsomes

1970 ◽  
Vol 120 (1) ◽  
pp. 1-13 ◽  
Author(s):  
R. Rodnight
Keyword(s):  
Sodium Chloride ◽  
Atp Hydrolysis ◽  
Sodium Dodecyl ◽  
Sodium Deoxycholate ◽  
Protein Ratio ◽  
Chemical Agents ◽  
Rapid Fall ◽  
Triton X ◽  
Hydrolysis Of ◽  
Sodium And Potassium

1. The effect of chemical agents on the turnover of the Na+-dependent bound phosphate and the simultaneous Na+-dependent hydrolysis of ATP by a membrane preparation from ox brain was studied at an ATP/protein ratio of 12.5pmol/μg. 2. The agents were added immediately after phosphorylation of the preparation in a medium containing 50mm-sodium chloride and 2.5μm-[γ-32P]ATP. 3. Concentrations of sodium chloride above 150mm, calcium chloride to 20mm and suramin to 1.4mm inhibited both phosphorylation and dephosphorylation and concomitantly slowed ATP hydrolysis. At 125mm-sodium chloride dephosphorylation and hydrolysis were slightly slowed without affecting phosphorylation. 4. Ethanol to 1.6m concentration inhibited dephosphorylation without affecting phosphorylation; the bound phosphate was increased and ATP hydrolysis slowed. 5. Ouabain to 4mm concentration partially inhibited ATP hydrolysis and caused a transient (1–2s) rise in bound phosphate followed by a rapid fall to a lower plateau value, which eventually declined to zero by the time ATP hydrolysis was complete. 6. Of the detergents examined Lubrol W, Triton X-100 and sodium deoxycholate had no significant effect on turnover. Sodium dodecyl sulphate and sodium decyl sulphate to 3.5mm and 20mm respectively completely inhibited turnover and ATP hydrolysis and stabilized the bound phosphate.

scholarly journals Metabolism of conjugated sterols in eggplant. Part 2. Phospholipid : steryl glucoside acyltransferase.

2008 ◽  
Vol 55 (1) ◽  
pp. 135-140 ◽  
Author(s):  
Anna Potocka ◽  
Jan Zimowski
Keyword(s):  
Divalent Cations ◽  
Solanum Melongena ◽  
Sodium Deoxycholate ◽  
Tween 20 ◽  
Acyl Donor ◽  
Kinetic Properties ◽  
Acyltransferase Activity ◽  
Steryl Glucoside ◽  
Membrane Bound ◽  
Triton X

A membrane-bound phospholipid : steryl glucoside acyltransferase from Solanum melongena leaves was partially purified and its specificity and molecular as well as kinetic properties were defined. Among the steryl glycosides tested (e.g. typical plant steryl glucosides, steryl galactosides and cholesteryl xyloside) the highest activity was found with cholesteryl glucoside, but some structurally related compounds such as sito- and stigmasteryl glucoside or galactoside as well as cholesteryl galactoside were also acylated, albeit at lower rates. The investigated enzyme was able to use all classes of phosphoglycerolipids (phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol) as an acyl source for biosynthesis of acylated steryl glucoside. Among them 1,2-dimirystoylphosphatidylic acid appeared to be the best acyl donor. Apart from phosphoglycerolipids, 1,2-diacylglycerols were also used as acyl donor for steryl glucoside acylation, although at a distinctly lower rate. The acyl moiety was transferred from the C-1 position of phospholipid molecule. The investigated acyltransferase activity was stimulated by 2-mercaptoethanol, Triton X-100, 1-monoacylglycerols and inhibited in the presence of divalent cations such as Ca(2+), Mn(2+), Zn(2+) or Co(2+), some lipids (MDGD, ceramide), detergents (Tween 20, 40, 60 and 80, Tyloxapol, sodium deoxycholate) and high ionic strength.

scholarly journals Methionine-Riboflavin Mixtures with Surfactants and Metal Ions Reduce Powdery Mildew Infection in Strawberry Plants

1998 ◽  
Vol 123 (6) ◽  
pp. 987-991 ◽  
Author(s):  
Shiow Y. Wang ◽  
Dean Der-Syh Tzeng
Keyword(s):  
Powdery Mildew ◽  
Foliar Application ◽  
Sodium Dodecyl ◽  
Free Radical Scavengers ◽  
Tween 20 ◽  
Sulfate Pentahydrate ◽  
Copper Sulfate Pentahydrate ◽  
Triton X ◽  
Powdery Mildew Infection ◽  
Β Carotene

Foliar application of a mixture of methionine (1 mm) and riboflavin (26.6 μm) reduced the severity of powdery mildew [Sphaerotheca macularia (Wallr. ex Fr.) Jacz. f. sp. fragariae] infection in `Earliglow' strawberry (Fragaria × ananassa Duch.) plants. Efficacy of this mixture on controlling powdery mildew infection was enhanced by supplements of copper, iron, and surfactants [sodium dodecyl sulfate (SDS), Triton X-100, Tween-20, or oxyalkylenemethylsiloxane (Silwet L-77)]. Free-radical scavengers (n-propyl gallate, thiourea) and antioxidants (α-tocopherol, β-carotene) reduced the efficacy of this mixture. Plants treated with a mixture of riboflavin (26.6 μm), d,l-methionine (1 mm), copper sulfate pentahydrate (1 mm), and surfactants (SDS or Silwet L-77 at concentrations of 0.05% to 0.1%) showed a decrease in powdery mildew infection. Results of this study suggest that treatment with a mixture of methionine and riboflavin is beneficial to strawberry plants and may serve as an alternative to fungicides for controlling powdery mildew.

scholarly journals Towards uterus tissue engineering: a comparative study of sheep uterus decellularisation

2020 ◽  
Vol 26 (3) ◽  
pp. 167-178 ◽  
Author(s):  
T T Tiemann ◽  
A M Padma ◽  
E Sehic ◽  
H Bäckdahl ◽  
M Oltean ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Sodium Dodecyl ◽  
Sodium Deoxycholate ◽  
Vascular Perfusion ◽  
Uterus Transplantation ◽  
Live Donors ◽  
Triton X

Abstract Uterus tissue engineering may dismantle limitations in current uterus transplantation protocols. A uterine biomaterial populated with patient-derived cells could potentially serve as a graft to circumvent complicated surgery of live donors, immunosuppressive medication and rejection episodes. Repeated uterine bioengineering studies on rodents have shown promising results using decellularised scaffolds to restore fertility in a partially impaired uterus and now mandate experiments on larger and more human-like animal models. The aim of the presented studies was therefore to establish adequate protocols for scaffold generation and prepare for future in vivo sheep uterus bioengineering experiments. Three decellularisation protocols were developed using vascular perfusion through the uterine artery of whole sheep uteri obtained from slaughterhouse material. Decellularisation solutions used were based on 0.5% sodium dodecyl sulphate (Protocol 1) or 2% sodium deoxycholate (Protocol 2) or with a sequential perfusion of 2% sodium deoxycholate and 1% Triton X-100 (Protocol 3). The scaffolds were examined by histology, extracellular matrix quantification, evaluation of mechanical properties and the ability to support foetal sheep stem cells after recellularisation. We showed that a sheep uterus can successfully be decellularised while maintaining a high integrity of the extracellular components. Uteri perfused with sodium deoxycholate (Protocol 2) were the most favourable treatment in our study based on quantifications. However, all scaffolds supported stem cells for 2 weeks in vitro and showed no cytotoxicity signs. Cells continued to express markers for proliferation and maintained their undifferentiated phenotype. Hence, this study reports three valuable decellularisation protocols for future in vivo sheep uterus bioengineering experiments.

Measurement of activin in biological fluids by radioimmunoassay, utilizing dissociating agents to remove the interference of follistatin

1996 ◽  
Vol 134 (4) ◽  
pp. 481-489 ◽  
Author(s):  
James R McFarlane ◽  
Lynda M Foulds ◽  
Angelique Pisciotta ◽  
David M Robertson ◽  
David M de Kretser
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Biological Fluids ◽  
Sodium Dodecyl ◽  
Sodium Deoxycholate ◽  
Male Rat ◽  
Tween 20 ◽  
Rat Serum ◽  
Level 3

McFarlane JR, Foulds LM, Pisciotta A, Robertson DM, de Kretser DM. Measurement of activin in biological fluids by radioimmunoassay, utilizing dissociating agents to remove the interference of follistatin. Eur J Endocrinol 1996;134:481–9. ISSN 0804–4643 Activin, a dimer of the β-subunits of inhibin, is a member of the transforming growth factor beta (TGF-β) superfamily of growth factors and has a widespread range of actions in a variety of tissues. The investigation of the physiology of activin action has been facilitated in recent years by the availability of immunoassays in addition to bioassays. Follistatin has been shown to bind to activin with a high affinity and therefore interferes in both radioimmunoassays and enzyme-linked immunosorbent assays (ELISAs). In this study we examined the effect of various surfactants and 1.4-dioxane on the measurement of activin in the presence of follistatin by radioimmunoassay. The addition of a combination of sodium deoxycholate, Tween 20 and sodium dodecyl sulphate removed the interference of follistatin in the radioimmunoassay. The measured content of activin in male rat serum, human male serum, human female serum and bovine follicular fluid rose from 3.29 to 4.15, < 0.48 to 2.87, 2.42 to 4.17 and 30.9 to 85.6 ng/ml, respectively, when assayed in the presence of the dissociating reagents. It was unclear whether the altered potencies were due to a dissociation of the follistatin/activin complex rather than the exposure of the epitope on activin recognized by the antiserum. Serum concentrations of activin were lower than those found in testicular cytosols, and after castration no change in serum activin levels was observed, suggesting that the testis does not contribute significantly to circulating activin levels. The use of the dissociating reagents in the radioimmunoassay will enable studies to be carried out that more accurately measure the activin content of various biological fluids, and thus lead to a greater understanding of the physiology of this growth factor. James McFarlane, Institute of Reproduction and Development, Level 3, Block E, Monash Medical Centre, 246 Clayton Road, Clayton, Victoria 3168, Australia

scholarly journals A new method for efficient detection of Cryptosporidium RNA by real-time reverse transcription-PCR with surfactants

2015 ◽  
Vol 15 (5) ◽  
pp. 1061-1068 ◽  
Author(s):  
Takahiro Sekikawa ◽  
Kosuke Toshiki
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
18S Rrna ◽  
Nonionic Surfactants ◽  
Sodium Dodecyl ◽  
Rrna Genes ◽  
Tween 20 ◽  
Acid Extraction ◽  
Rt Pcr ◽  
Triton X

Cryptosporidium is one of the most common causes of waterborne diseases worldwide. Its oocysts possess a robust wall that is extremely resistant to the chlorine used for potable water disinfection. The current procedures of nucleic acid extraction and purification, such as the freeze–thaw (F/T) method and the commercial kits, are time consuming and expensive. To this end, a surfactant extraction treatment (SET) was developed as a method to extract nucleic acids from Cryptosporidium using only surfactants. The use of 18S rRNA improves the sensitivity of Cryptosporidium detection for real-time polymerase chain reaction (PCR), because 18S rRNA molecules are constitutively present in high copy numbers. Therefore, we applied SET to the detection of Cryptosporidium 18S rRNA using reverse transcription (RT)-PCR for the first time. RT-PCR was inhibited by 0.01% of the anionic surfactant sodium dodecyl sulfate (SDS), whereas the inhibition did not occur with 5% of the nonionic surfactants Tween 20, Triton X-100, Tween 80, and Triton X-114. However, the nonionic surfactants could not completely suppress the inhibition induced by 0.1% SDS. We successfully extracted 18S rRNA genes from oocysts by SET without the F/T method and detected them by real-time RT-PCR.

scholarly journals Effect of detergent type on the performance of liver decellularized extracellular matrix-based bio-inks

2021 ◽  
Vol 12 ◽  
pp. 204173142199709
Author(s):  
Wonwoo Jeong ◽  
Min Kyeong Kim ◽  
Hyun-Wook Kang
Keyword(s):  
Extracellular Matrix ◽  
Sodium Dodecyl ◽  
Great Influence ◽  
Ammonium Hydroxide ◽  
Sodium Deoxycholate ◽  
Porcine Liver ◽  
Decellularized Extracellular Matrix ◽  
Cytochrome P450 Activity ◽  
Primary Mouse ◽  
Triton X

Decellularized extracellular matrix-based bio-inks (dECM bio-inks) for bioprinting technology have recently gained attention owing to their excellent ability to confer tissue-specific functions and 3D-printing capability. Although decellularization has led to a major advancement in bio-ink development, the effects of detergent type, the most important factor in decellularization, are still unclear. In this study, the effects of various detergent types on bio-ink performance were investigated. Porcine liver-derived dECM bio-inks prepared using widely used detergents, including sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC), Triton X-100 (TX), and TX with ammonium hydroxide (TXA), were characterized in detail. SDS and SDC severely damaged glycosaminoglycan and elastin proteins, TX showed the lowest rate of decellularization, and TXA-based dECM bio-ink possessed the highest ECM content among all bio-inks. Differences in biochemical composition directly affected bio-ink performance, with TXA-dECM bio-ink showing the best performance with respect to gelation kinetics, intermolecular bonding, mechanical properties, and 2D/3D printability. More importantly, cytocompatibility tests using primary mouse hepatocytes also showed that the TXA-dECM bio-ink improved albumin secretion and cytochrome P450 activity by approximately 2.12- and 1.67-fold, respectively, compared with the observed values for other bio-inks. Our results indicate that the detergent type has a great influence on dECM damage and that the higher the dECM content, the better the performance of the bio-ink for 3D bioprinting.

scholarly journals Surfactant toxicity to Artemia franciscana and the influence of humic acid and chemical composition

2016 ◽  
Vol 13 (3) ◽  
pp. 507 ◽  
Author(s):  
Rachel D. Deese ◽  
Madeline R. LeBlanc ◽  
Robert L. Cook
Keyword(s):  
Humic Acids ◽  
Cetylpyridinium Chloride ◽  
Sodium Dodecyl ◽  
Soxhlet Extraction ◽  
Artemia Franciscana ◽  
Aquatic Environments ◽  
Chemically Modified ◽  
Alkyl Groups ◽  
Modified Humic Acids ◽  
Triton X

Environmental context Surfactants, a pollutant class routinely introduced into aquatic environments, can be toxic to a variety of species. It is thus important to understand how surfactants’ toxicity is influenced by their interactions with other environmental constituents, including natural organic matter. We report the changes in toxicity of three surfactants to brine shrimp in the presence of unmodified and chemically modified humic acids. Abstract Surfactants can be extremely toxic to aquatic species and are introduced to the environment in a variety of ways. It is thus important to understand how other environmental constituents, in this case humic acids (HAs), may alter the toxicity of anthropogenic surfactants. Hatching and mortality assays of Artemia Franciscana were performed for three different toxic surfactants: Triton X-100 (Tx-100, non-ionic), cetylpyridinium chloride (CPC, cationic) and sodium dodecyl sulfate (SDS, anionic). HAs of varying composition and concentrations were added to the assays to determine the toxicity mitigating ability of the HAs. Tx-100 had a significant toxic effect on Artemia mortality rates and HAs from terrestrial sources were able to mitigate the toxicity, but an aquatic HA did not. CPC and SDS limited hatching success of the Artemia and, as HAs were added, the hatching percentages increased for all HA sources, indicating toxicity mitigation. In order to determine which functional groups within HAs were responsible for the interaction with the surfactants, the HAs were chemically modified by: (i) bleaching to reduce aromatics, (ii) Soxhlet extraction to reduce lipids and (iii) acid hydrolysis to reduce O- and N-alkyl groups. Although most of the modified HAs had some toxicity mitigating ability for each of the surfactants, there were two notable differences: (1) the lipid-extracted HA did not reduce the toxicity of Tx-100 and (2) the bleached HA had a lower toxicity mitigating ability for CPC than the other modified HAs.

Purification and Characterization of Thermostable α-Amylase from Soil Bacterium Bacillus sp.

2021 ◽  
Vol 28 ◽  
Author(s):  
Barış Enez
Keyword(s):  
Metal Ions ◽  
Amylase Activity ◽  
Low Cost ◽  
Sodium Dodecyl ◽  
Tween 20 ◽  
Bacillus Sp ◽  
Wide Range ◽  
Bacterium Bacillus ◽  
Triton X

Background: Amylases are used in several industrial and biotechnological sectors, including those producing textiles, detergents, paper and bakery products. Objective: This study aimed to purify an industrially important α-amylase from Bacillus sp. For this purpose, a single and rapid α-amylase purification was performed using the starch affinity method. Methods: Characterization of the purified enzyme was determined by investigating temperature, pH stability, detergents, and metal ions. Results: The purification coefficient of 29.8-fold with a yield of 9.2% was found. The molecular weight of the purified α-amylase was determined to be 53 kDa by SDS-PAGE, and thermostability was confirmed with 100% activity at 30ºC and 40ºC after 1 h. The purified enzyme was stable over a wide range of pH values, with optimum activity at pH 6.0, 7.0 and 8.0 after 2 h. The study also investigated the effects of the metal ions and detergents on the purified amylase and found that Mg2+ and Ca2+ ions were the activators of the enzyme, while Zn2+, Co2+ and Na+ ions decreased the activity. Furthermore, Hg2+ indicated complete inhibition of amylase activity. The detergents Triton X-100 and Tween 20 increased the α-amylase activity, while sodium dodecyl sulfate inhibited the activity. Conclusion: The purified α-amylase obtained from Bacillus sp. is considered to be environmentally friendly, can be processed in a short time, and has a low cost.

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