sodium-dodecyl-sulfate and sarkosyl

Sodium dodecyl sulfate (SDS) is a detergent used as a strong denaturant of proteins in gel electrophoresis. It has previously been shown that certain hyperstable, also known as kinetically stable, proteins are resistant to SDS and thus require heating for their denaturation in the presence of SDS. Because of its high denaturing strength, relatively few proteins are resistant to SDS thereby limiting the current use of SDS-PAGE for identifying hyperstable degradation-resistant proteins. In this study, we show that sarkosyl, a milder detergent than SDS, is able to identify proteins with moderately high kinetic stability that lack SDS-resistance. Our assay involves running and subsequently comparing boiled and unheated protein samples containing sarkosyl, instead of SDS, on PAGE gels and identifying subsequent differences in protein migration. Our results also show that sarkosyl and SDS may be combined in PAGE experiments at varying relative percentages to obtain semi-quantitative information about a protein's kinetic stability in a range inaccessible by probing through native- or SDS-PAGE. Using protein extracts from various legumes as model systems, we detected proteins with a range of protein stability from nearly SDS-resistant to barely sarkosyl resistant.

Topics: Detergents; Electrophoresis, Polyacrylamide Gel; Kinetics; Molecular Structure; Protein Stability; Proteins; Sarcosine; Sodium Dodecyl Sulfate

Shampoo and cleansers containing anionic surfactants including sodium dodecyl sulphate (SDS) often cause pruritus in humans. Daily application of 1-10% SDS for 4 days induced hind-paw scratching (an itch-related behaviour) in a concentration-dependent manner, and 10% SDS also caused dermatitis, skin dryness, barrier disruption, and an increase in skin surface pH in mice. SDS-induced scratching was inhibited by the opioid receptor antagonist naloxone and the H histamine receptor antagonist terfenadine. Mast-cell deficiency did not inhibit SDS-induced scratching, although it almost completely depleted histamine in the dermis. Treatment with SDS increased the histamine content of the epidermis, but not that of the dermis. SDS treatment increased the gene expression and post-translation processing of L-histidine decarboxylase in the epidermis. The present results suggest that repeated application of SDS induces itch through increased production of epidermal histamine, which results from an increase in the gene expression and post-translation processing of L-histidine decarboxylase.

Topics: Animals; Behavior, Animal; Disease Models, Animal; Enzyme Induction; Epidermis; Histamine; Histamine H1 Antagonists; Histidine Decarboxylase; Male; Mast Cells; Mice, Inbred ICR; Narcotic Antagonists; Protein Processing, Post-Translational; Pruritus; Sarcosine; Signal Transduction; Sodium Dodecyl Sulfate; Surface-Active Agents; Time Factors; Up-Regulation

Oral drug delivery offers an attractive method of needle-free drug administration. Unfortunately, oral delivery is often hampered by the poor permeability of drugs across the intestinal epithelium. Although several single chemical permeation enhancers have been shown to alleviate permeability difficulties, this often occurs at the expense of safety. This in vitro study demonstrates the use of binary and ternary combinations of permeation enhancers to create synergistic enhancer formulations (SEFs) that offer a high level of potency while inducing very little toxicity in Caco-2 cells. Although relatively rare in the explored formulation space, SEFs were abundant enough to significantly increase the repertoire of permeation enhancers that are safe and effective in vitro. The most promising enhancers from the binary study led to easily identifiable ternary SEFs, thus increasing the efficiency of the discovery process. Some of the best performers of the study included binary combinations of hexylamine and chembetaine and ternary combinations of sodium laureth sulfate, decyltrimethyl ammonium bromide, and chembetaine, all at a total concentration of 0.1% (w/v). Furthermore, several SEFs were shown to be capable of increasing mannitol and 70 kDa dextran permeability across Caco-2 monolayers 15- and 8-fold, respectively. These results encourage further exploration of several leading formulations for in vivo applications in oral drug delivery.

Topics: Adjuvants, Pharmaceutic; Administration, Oral; Amines; Bicyclic Monoterpenes; Caco-2 Cells; Cell Membrane Permeability; Cell Survival; Cholic Acids; Decanoic Acids; Dextrans; Drug Combinations; Drug Synergism; Electric Impedance; Epithelial Cells; Humans; Intestinal Absorption; Intestinal Mucosa; Mannitol; Pyrrolidinones; Quaternary Ammonium Compounds; Sarcosine; Sodium Dodecyl Sulfate; Surface-Active Agents; Terpenes

This report focuses on the development of a method for chemically induced enhancement of cell elution and recovery from cotton swabs. The method exploits the exclusive use of detergents for intact cell removal, and can be utilized in conjunction with, or to circumvent, conventional differential extraction (DE). Samples treated with Sarkosyl (54.4 +/- 1.8%) and sodium dodecyl sulfate (SDS) (78.5 +/- 0.7%) yielded higher sperm cell recoveries than a conventional DE buffer (39.4 +/- 2.1%). The results indicated that the choice of detergent affected sperm cell yield, with anionic detergents having the greatest effect. Storage time of samples affected the concentration of detergent required for optimal sperm cell recovery, longer times requiring increased detergent concentrations. In addition, the extent of sperm cell lysis by proteinase K digestion was evaluated. The results indicate that the exclusive use of SDS enhances the release of sperm and epithelial cells from a cotton swab as compared with DE buffer, providing for a more effective DNA analysis.

Topics: Cell Separation; DNA Fingerprinting; Female; Forensic Genetics; Humans; Male; Rape; Sarcosine; Sodium Dodecyl Sulfate; Spermatozoa

This research reports the sensitivity of a clinical isolate of Enterococcus faecalis to sodium N-lauroylsarcosinate (sarkosyl) and sodium dodecyl sulfate (SDS), as well as the efficiency of these detergents in curing the strain. Compared to Escherichia coli, Enterococcus faecalis was very sensitive to both detergents, with minimum inhibitory concentrations (MIC) for the latter being 100 times lower than for Escherichia coli. The clinical isolate of Enterococcus faecalis used in this study exhibited plasmid-borne resistance to kanamycin (MIC 2 mg/ml) and tetracycline (MIC 50 mug/ml); 3% curing was observed after growth in the presence of sarkosyl but no curing was observed after growth in the presence of either SDS or acridine orange. In contrast, 35% curing of plasmid-bearing Escherichia coli was observed after growth in the presence of either SDS or acridine orange, but none was observed after growth in the presence of sarkosyl.

Topics: Acridine Orange; Anti-Bacterial Agents; Enterococcus faecalis; Escherichia coli; Kanamycin Resistance; Microbial Sensitivity Tests; Plasmids; Sarcosine; Sodium Dodecyl Sulfate; Tetracycline Resistance

The mechanisms of herpes simplex virus (HSV) inactivation by sodium lauryl sulfate (SLS) and n-lauroylsarcosine (LS), two anionic surfactants with protein denaturant potency, have been evaluated in cultured cells. Results showed that pretreatment of HSV type 1 (HSV-1) strain F and HSV-2 strain 333 with either surfactant inhibited, in a concentration- and time-dependent manner, their infectivities on Vero cells. SLS was a more potent inhibitor of HSV-2 strain 333 infectivity than LS with respect to the concentration (4.8-fold lower) and time (2.4-fold shorter) required to completely inactivate the virus. No inhibition of both herpesvirus strains infectivities was observed when Vero cells were pretreated with either surfactant. LS prevented the binding of HSV-2 strain 333 to cells without affecting the stable attachment and the rate of penetration into cells, whereas SLS exerted the opposite effect. Both SLS and LS inhibited, in a concentration-dependent manner, the HSV-2 strain 333-induced cytopathic effect, probably by affecting newly synthesized virions that come into contact with surfactant molecules present in culture medium. The pretreatment of HSV-2 strain 333 with specific combinations of SLS and LS concentrations inhibited the viral infectivity in a synergistic manner and resulted in only a small increase in their toxicities for exponentially growing Vero cells compared with that caused by each compound alone. Taken together, these results suggest that SLS and LS, alone or combined, could represent potent candidates as microbicides in topical vaginal formulations to prevent the transmission of herpes and possibly other pathogens that cause sexually transmitted diseases, including human immunodeficiency virus type 1.

Topics: Animals; Cell Survival; Chlorocebus aethiops; Herpesvirus 1, Human; Herpesvirus 2, Human; Receptors, Virus; Sarcosine; Sodium Dodecyl Sulfate; Surface-Active Agents; Time Factors; Vero Cells; Viral Plaque Assay; Viral Proteins

The microbicidal efficacies of two anionic surfactants, sodium lauryl sulfate (SLS) and n-lauroylsarcosine (LS), were evaluated in cultured cells and in a murine model of herpes simplex type 2 (HSV-2) intravaginal infection. In vitro studies showed that SLS and LS were potent inhibitors of the infectivity of HSV-2 strain 333. The concentrations of SLS which inhibit viral infectivity by 50% (50% inhibitory dose) and 90% (90% inhibitory dose) were 32.67 and 46.53 microM, respectively, whereas the corresponding values for LS were 141.76 and 225.30 microM. In addition, intravaginal pretreatment of mice with thermoreversible gel formulations containing 2.5% SLS or 2.5% LS prior to the inoculation of HSV-2 strain 333 completely prevented the development of genital herpetic lesions and the lethality associated with infection. Of prime interest, no infectious virus could be detected in mouse vaginal mucosa. Both formulations still provided significant protection when viral challenge was delayed until 1 h after pretreatment. Finally, intravaginal application of gel formulations containing 2.5% SLS or 2.5% LS once daily for 14 days to rabbits did not induce significant irritations to the genital mucosa, as demonstrated from macroscopic and histopathologic examinations. These results suggest that thermoreversible gel formulations containing SLS or LS could represent potent and safe topical microbicides for the prevention of HSV-2 and possibly other sexually transmitted pathogens, including human immunodeficiency virus.

Topics: Administration, Topical; Analysis of Variance; Animals; Anti-Infective Agents; Area Under Curve; Chemistry, Pharmaceutical; Chlorocebus aethiops; Detergents; Female; Gels; Herpes Genitalis; Herpesvirus 2, Human; Mice; Mice, Inbred BALB C; Rabbits; Sarcosine; Sexually Transmitted Diseases; Sodium Dodecyl Sulfate; Surface-Active Agents; Vagina; Vero Cells

Cytoplasmic membranes of Streptococcus mutans GS-5 were extracted by incubating cells with 1% sodium lauroyl sarcosinate for 20 min at room temperature. The profiles of membrane proteins were determined by SDS-PAGE and isoelectric focusing. The effect of chlorhexidine digluconate on cell membranes was studied after treating the extracted proteins for 30 min with the drug at final concentrations of 0.05 and 0.2%. Chlorhexidine caused selective reduction in the intensity of the membrane proteins. Five densely staining bands with molecular weights of 24.2, 19.6, 18.1, 17.6 and 16.4 kD were obviously diminished. Isoelectric focusing indicated that chlorhexidine preferably precipitated acidic cytoplasmic proteins (pI 4.0-4.92). Our results are consistent with the hypothesis that the mode of action of this drug is mainly attributed to its hydrophilic property. Chlorhexidine, being cationic, may interact with bacteria by attraction to negative-charge membrane components.

Topics: Adenosine Triphosphatases; Anti-Infective Agents, Local; Cell Membrane; Chemical Precipitation; Chlorhexidine; Cytoplasm; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Isoelectric Focusing; Membrane Proteins; Molecular Weight; Sarcosine; Sodium Dodecyl Sulfate; Staining and Labeling; Streptococcus mutans; Surface-Active Agents

When primary cultures of hepatocytes are exposed to media containing fetal bovine serum (FBS) there is a rapid decrease in levels of tissue-specific mRNAs such as albumin mRNA. We used Northern blot analysis to examine mRNA levels in cultured hepatocytes, and characterized the factor in FBS that significantly reduces the steady state albumin mRNA level. Neonatal bovine serum or serum derived from platelet-poor calf plasma proved as potent as did FBS, but commercial bovine serum albumin did not exhibit this inhibitory activity. Inhibitory activity of FBS was not removed by moderate heat treatment, dialysis, or extraction with organic solvents. However, incubation of FBS with a highly anionic detergent such as 0.1% sodium dodecyl sulfate or N-lauroyl sarcosine, followed by extensive dialysis, resulted in sera that did not inhibit expression of albumin mRNA. These sera supported cell attachment and seemed non-toxic toward the cells. Ammonium sulfate fractionation of FBS showed the activity was present in the 45 to 70% fraction, and trypsin digestion destroyed the inhibitory activity. Gel exclusion chromatography gave a molecular weight of 60,000 to 70,000. Fractionation of serum proteins by DEAE-Sephacel or Cibacron blue-agarose showed enrichment for albumin in the most active fractions. Interestingly, metabolic labeling of secreted and cellular proteins with 35S-methionine and cysteine showed no significant difference between hepatocytes maintained for 2 days beforehand in serum-free or serum-supplemented media, and no difference between detergent-treated FBS and control FBS.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Biological Factors; Cells, Cultured; Detergents; Gene Expression Regulation; Liver; Male; Molecular Weight; Plasma; Rats; Rats, Sprague-Dawley; RNA, Messenger; Sarcosine; Serum Albumin; Sodium Dodecyl Sulfate

Virus recovery from poliovirus-seeded waste water was attempted after treatment of the samples with ionic detergents in the presence of acoustic energy. Viral titers strongly fluctuated depending upon concentration and chemical structure of the detergent as well as upon the dissolved organic content of the aqueous samples. Supplementation of the cellular monolayers with an additional amount of cell culture medium or with a nonionic detergent in a subcytotoxic concentration 48 h after inoculation partly induced cytopathic effects in silent dilution steps. Since, in the presence of the same detergent, viral recovery rates varied with the type of waste water, titer-conditioning activity of detergents was suggested to depend upon their effective critical micellization concentration. Especially for virus recovery from concentrated waste water, treatment with strongly disruptive detergents such as sodium dodecylsulfate revealed to be much more efficient than with the less hydrophilic sodium N-laurylsarcosine, sodium glycodeoxycholate or lauryldimethylamine-oxide, whereas the opposite seemed to be the case if virus recovery from filtrate samples were to be optimized. The mechanisms by which viral particles become infectious for the cell appear to be triggered by general principles of equilibrium thermodynamics, which means that interactions between viral surface proteins and cellular receptor molecules seem to reflect the tendency of these reagents to assume the energetically most favored orientation.

Topics: Animals; Cell Line; Chlorocebus aethiops; Cytopathogenic Effect, Viral; Detergents; Dimethylamines; Glycodeoxycholic Acid; Kidney; Micropore Filters; Poliovirus; Sarcosine; Sewage; Sodium Dodecyl Sulfate; Water Microbiology

Most commercial dentifrices contain sodium lauryl sulphate, which oxidizes upon storage. The effects of aged sodium lauryl sulphate and its oxidative breakdown products on the conversion of amorphous calcium phosphate to hydroxyapatite were studied in vitro by a pH drop method. Hydroxyapatite was identified from its X-ray diffraction pattern. With storage time, the concentration of dodecanol [CH3(CH2)11OH], a breakdown product of sodium lauryl sulphate, increased. The storage dodecanol-containing sodium lauryl sulphate accelerated the conversion of amorphous calcium phosphate to hydroxyapatite. Dodecanol mixed with sodium lauryl sulphate accelerated the conversion when added both before and after initial formation of amorphous calcium phosphate. A stored commercial dentifrice also accelerated the conversion of amorphous calcium phosphate to hydroxyapatite. It was found that the concentration of dodecanol increased 2-fold over a 2-month period.

Topics: 1-Butanol; Butanols; Calcium; Calcium Phosphates; Chemical Precipitation; Chromatography, Gas; Detergents; Dodecanol; Drug Storage; Durapatite; Ethanol; Hydrogen-Ion Concentration; Hydroxyapatites; Methanol; Oxidation-Reduction; Phosphates; Sarcosine; Sodium Dodecyl Sulfate; Solubility; Spectrophotometry, Atomic; Spectrophotometry, Ultraviolet; Toothpastes; X-Ray Diffraction

Borrelia burgdorferi and Borrelia hermsii were treated with the following detergents: sodium dodecyl sulphate (SDS), N-lauryl sarcosine (Sarkosyl) and Triton X-100, and the soluble and insoluble fractions obtained after each detergent treatment were examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Sarkosyl as well as SDS solubilized all Borrelia proteins, whereas Triton X-100 treatment selectively solubilized the majority of other borrelial proteins, leaving two proteins of 41 kDa and 66 kDa in Borrelia burgdorferi and one protein of 39 kDa in Borrelia hermsii associated with the Triton X-100 insoluble fraction.

Topics: Bacterial Proteins; Blotting, Western; Borrelia; Borrelia burgdorferi Group; Detergents; Electrophoresis, Polyacrylamide Gel; Molecular Weight; Octoxynol; Polyethylene Glycols; Sarcosine; Sodium Dodecyl Sulfate

The effect of detergent type, pH and temperature during lysis on the DNA elution dose response was studied under non-winding conditions in exponentially growing, plateau-phase and synchronous S-phase CHO cells. Lysis with sodium-N-laurylsarcosine (NLS) increased the DNA elution rate and resulted in higher DNA elution for the same absorbed radiation dose than lysis with sodium dodecyl sulphate (SDS). This increase in elution caused a reduction in the shoulder width of the DNA elution dose-response curve, but did not significantly affect the final slope. One hour incubation at elevated temperatures (60 degrees C) during lysis either with NLS or SDS further increased DNA elution. Under these conditions DNA elution dose-response curves with a small or zero shoulder were obtained with exponentially growing, plateau-phase or synchronized S-phase cells. DNA elution was reduced to about 50 per cent of the controls when the pH of the SDS lysis solution was adjusted from 9.6 to 7.6. This effect was observed in cells that were lysed at room temperature, as well as in cells lysed at 60 degrees C. When NLS was used for lysis, a similar reduction in pH did not alter the DNA elution dose-response curve at either lysis temperature. Based on these results it is suggested that the shoulder observed in the DNA elution dose-response curve reflects partial separation of DNA from associated proteins. A direct and unconditional correlation of the DNA filter elution behaviour, as observed under non-unwinding conditions, with the induction of DNA dsb may thus not always be justified. Caution is required when elution results are used to establish correlations between the level of induction of DNA dsb and cell killing.

Topics: Animals; Cell Cycle; Cell Line; Cricetinae; Cricetulus; Detergents; DNA; Dose-Response Relationship, Drug; Female; Hydrogen-Ion Concentration; Ovary; Sarcosine; Sodium Dodecyl Sulfate; Temperature

The cell wall of the biflagellate alga Chlamydomonas reinhardtii is a multilayered, extracellular matrix composed of carbohydrates and 20-25 polypeptides. To learn more about the forces responsible for the integrity of this cellulose-deficient cell wall, we have begun studies to identify and characterize the framework of the wall and to determine the effects of the cell wall-degrading enzyme, lysin, on framework structure and protein composition. In these studies we used walls released into the medium by mating gametes. When isolated shed walls are degraded by exogenously added lysin, no changes are detected in the charge or molecular weight of the 20-25 wall proteins and glycoproteins when analyzed on one- and two-dimensional polyacrylamide gels, which suggests that degradation of these shed walls is due either to cleavage of peptide bonds very near the ends of polypeptides or that degradation occurs via a mechanism other than proteolysis. Incubation of walls with Sarkosyl-urea solutions removes most of the proteins and yields thin structures that appear to be the frameworks of the walls. Analysis by polyacrylamide gel electrophoresis shows that the frameworks are highly enriched in a polypeptide of Mr 100,000. Treatment of frameworks with lysin leads to their degradation, which indicates that this part of the wall is a substrate for the enzyme. Although lysin converts the Mr 100,000 polypeptide from an insoluble to a soluble form, there is no detectable change in Mr of the framework protein. Solubilization in the absence of lysin requires treatment with SDS and dithiothreitol at 100 degrees C. These results suggest that the Chlamydomonas cell wall is composed of two separate domains: one containing approximately 20 proteins held together by noncovalent interactions and a second domain, containing only a few proteins, which constitutes the framework of the wall. The result that shed walls can be solubilized by boiling in SDS-dithiothreitol indicates that disulfide linkages are critical for wall integrity. Using an alternative method for isolating walls from mechanically disrupted gametes, we have also shown that a wall-shaped portion of these unshed walls is insoluble under the same conditions in which shed walls are soluble. One interpretation of these results is that wall release during mating and the wall degradation that follows may involve distinct biochemical events.

Topics: Carbohydrates; Cell Wall; Chlamydomonas; Dithiothreitol; Extracellular Matrix; Mucoproteins; Peptides; Sarcosine; Sodium Dodecyl Sulfate; Stress, Mechanical; Urea

Topics: Animals; Detergents; Gingiva; Rats; Rats, Inbred Strains; Sarcosine; Soaps; Sodium Dodecyl Sulfate; Surface-Active Agents

Topics: Dentifrices; Detergents; Gingiva; Humans; Oral Hygiene; Sarcosine; Soaps; Sodium Dodecyl Sulfate; Surface-Active Agents

Topics: Adolescent; Detergents; Female; Gingiva; Humans; Male; Sarcosine; Soaps; Sodium Dodecyl Sulfate; Surface-Active Agents; Toothpastes

Topics: Cells, Cultured; Detergents; Embryo, Mammalian; Fibroblasts; Humans; Sarcosine; Soaps; Sodium Dodecyl Sulfate; Surface-Active Agents

Topics: Animals; Cell Nucleus; Centrifugation, Density Gradient; Chromatin; DNA, Neoplasm; Electrophoresis, Agar Gel; Electrophoresis, Polyacrylamide Gel; Friend murine leukemia virus; Leukemia, Experimental; Mice; Micrococcal Nuclease; Nucleosomes; Octoxynol; Polyethylene Glycols; Sarcosine; Sodium Dodecyl Sulfate

A whole cell lysate preparation of the Escherichia coli strain F11(P155) was treated with the following agents: heat (60 degrees C for 30 minutes), pronase, lipase, amylase, formalin (0.1%), sodium lauryl sulfate (0.05%) and sarkosyl NL30 (0.05%). Except for the amylase treatment, the treated whole cell lysate was inactivated when tested in rabbit gut loops or on Chinese hamster ovary cells. The preparations treated with heat or formalin could be used to produce a neutralizing antiserum or could remove the neutralizing capacity of an anti-F11(P155) serum. Finally, the attempt to demonstrate enterotoxicity with the cytoplasmic membrane of the bacterial cell failed.

Topics: Amylases; Animals; Cell Membrane; Cricetinae; Detergents; Enterotoxins; Escherichia coli; Formaldehyde; Hot Temperature; Immune Sera; Lauric Acids; Lipase; Pronase; Rabbits; Sarcosine; Sodium Dodecyl Sulfate