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At least 19 recordsLinked to original sources

PubMed1986

Evaluation of the cytotoxicity and genotoxicity of the spermicides nonoxynol-9 and octoxynol-9.

The cytotoxic and genotoxic potentials of the spermicidal agents, nonoxynol-9 (N-9) and octoxynol-9 (0-9), were evaluated in in vitro test systems using rat liver cells. N-9 was also tested for the induction of sperm abnormalities in mice. Dose-related cytocidal effects were seen after the addition of N-9 and O-9 to the culture medium for 24 h. The mean concentrations of N-9 and O-9 necessary to decrease the number of viable cells by 50% (LC50) were 24 and 43 micrograms/ml of media, respectively. The spermicides neither induced DNA repair in freshly isolated hepatocytes, nor caused any mutations at HGPRT locus in the T51B rat liver cell line. There was also a lack of malignant transformation response in the low-calcium assay. Further, the germinal cells of mice remained unaffected by N-9.

Animals↗

PubMed1987

Immunotoxicology studies on octoxynol-9 and nonoxynol-9 in mice.

In a double-blind study, mice were injected intraperitoneally with 0.2 ml 0.2% octoxynol-9 (O-9), 0.2 ml 0.2% nonoxynol-9 (N-9), or 0.2 ml saline (control) daily for 24 days. Another control group received no treatment. All mice were immunized twice with sheep red blood cells (SRBC) and bled by caudal incision. Mice receiving N-9 lost weight (P less than 0.02), had smaller livers (P less than 0.05), and showed enlarged spleens (P less than 0.05). The N-9-treated mice did not differ from either control group in the primary or secondary anti-SRBC responses, leukocyte (WBC) counts, or in the sizes of the kidneys, hearts, lungs, or thymuses. Mice receiving O-9 showed no significant differences from either control group in any of these tests. Serum immunoglobulin M (IgM) and immunoglobulin G (IgG) levels were similar in mice treated with O-9, N-9, or saline. All 3 groups had higher levels of both classes of immunoglobulins on day 16 than did untreated controls. This study shows that O-9, given to mice in doses 3 times that used by humans, is nontoxic, whereas the same dose of N-9 has minor deleterious effects.

Animals↗

PubMed1985

Effect of nonionic surfactants on aqueous primidone suspensions.

The following interactions between the soluble surfactant, octoxynol 9, and the very slightly soluble, finely powdered drug, primidone, in aqueous suspension were investigated: adsorption/desorption of the surfactant, micellar solubilization of the drug, and deflocculation of its particles. The last effect, measured by the sedimentation volume of the suspensions, was also investigated for other octoxynols. The adsorption of octoxynol 9 on solid primidone was proportional to the equilibrium surfactant concentration up to the critical micelle concentration. It leveled off at higher concentrations, reaching saturation at completion of a close-packed surfactant monolayer. The adsorption was essentially completely reversible. The solubility of primidone in water was very slight; its micellar solubilization was even less extensive. The sedimentation volume of primidone suspensions decreased with increasing equilibrium concentration of octoxynol 9 and began to level off at the critical micelle concentration of 0.018%. At about twice that concentration, the sedimentation volume became almost constant, but reached its lowest value only at less than or equal to 0.5%. Slow rotation of suspensions prior to sedimentation promoted flocculation and higher sedimentation volumes between 0.01 and 0.03% octoxynol 9. Octoxynols with higher hydrophilic-lipophilic balance than octoxynol 9 produced considerably larger sedimentation volumes at comparable concentrations, due to a lower surface activity and a lesser tendency to adsorb on primidone.

Micelles↗

PubMed1976

Effect of inorganic additives on solutions of nonionic surfactants III: CMC's and surface properties.

Continuing work on the interaction of inorganic additives with nonionic surfactants in aqueous solution dealt with their effect on the CMC and surface tension. The surfactants were octoxynol and polyoxyethylated oleyl alcohol, containing an average of 9.5 and 10 ethylene oxide units, respectively. Their CMC values were lowered by most electrolytes studied, representing salting out of the surfactants. The steepest reductions in the CMC were produced by the nitrates of sodium and potassium, which had been found to lower the cloud points of nonionic surfactants, salting them out because of the inability of their cations to form complexes with the ether oxygen linkages of the polyoxyethylene moieties. However, even electrolytes with cations such as hydrogen, lithium, calcium, nickel, lead, and aluminum capable of forming complexes with the ether oxygens, thereby increasing the cloud points of the surfactants, lowered their CMC values. In the presence of increasing concentrations of the latter electrolytes, the CMC values frequently went through minima and approached the CMC of the surfactant in the absence of additives. Increases in the CMC over the entire range of additive concentrations investigated were produced by cadmium nitrate for octoxynol, urea for polyoxyethylated oleyl alcohol, and magnesium nitrate for both. Net increases in the plateau or micellar surface tension of polyoxyethylated oleyl alcohol, i.e., in the constant surface tension of surfactant solutions above the CMC, were brought about by the nitrates of cadmium, aluminum, and magnesium at low concentrations only and by urea at all concentrations. This increase is interpreted as salting in. The area per surfactant molecule adsorbed at the air-water interface was reduced by all added electrolytes. Urea caused no such reduction.

Adsorption↗

PubMed1992

Virus inactivation of fresh frozen plasma by a solvent detergent procedure: biological results.

In order to increase the safety of blood products, we have developed a procedure for the virus inactivation of fresh frozen plasma. Several batches have been prepared and with the first 10 batches, each of them composed of 60 litres of plasma, we have determined a set of biological parameters. Virus inactivation was realised using TnBP (1%) and Octoxynol 9 (1%). After their elimination with castor oil using chromatography on insolubilized C18 resin, glycine was added and the pH of the plasma was adjusted to 7.4. Plastic bags were aseptically filled with a mean volume of 200 ml of plasma. The mean levels of coagulation factors were all over 0.7 U/ml and their recovery from initial plasma was nearly the same as total protein except for factor VIII:C. The net loss in factor VIII:C was 16%, when including the dilution of plasma. In vivo and in vitro tests demonstrated that in the final product there were no activated factors. As in fresh frozen plasma, the protein concentration was over 50 g/l and the potassium level lower than 5 mmol/l. According to these results, virus-inactivated plasma has the same qualities of fresh frozen plasma and could now replace it.

Blood Coagulation Factors↗

PubMed1977

Effect of symmetrical tetraalkylammonium salts on cloud point on nonionic surfactants.

The salting in and salting out of the nonionic surfactant octoxynol NF by halides of ammonium and the four lowest symmetrical tetraalkylammonium cations were investigated by measuring their effect on the cloud point at various salt concentrations. The chloride anion tended to salt the surfactant out, lowering its cloud point. The iodide anion tended to salt it in, raising the cloud point, while the bromide ion had no pronounced effect. The ammonium and tetramethylammonium cations tended to lower the cloud point, the latter more extensively than the former. The tetraethylammonium cation had no pronounced effect, while the tetra-n-propylammonium and tetra-n-butylammonium cations tended to raise the cloud point, the latter more extensively than the former. The salt effect on the cloud point generally increased with increasing salt concentration. Tetramethylammonium chloride was the most efficient salt in lowering the cloud point (by 4 degrees at the 0.10-molal level and by 20 degrees at the 1.00-molal level), while tetrabutylammonium halides were the most efficient salts in raising the cloud point (by 18-19 degrees at the 0.10-molal level for chloride and bromide). Cloud point increased were attributed to the formation of mixed micelles or to hydrotropy. The salting-out efficiency of tetramethylammonium chloride and bromide was ascribed to their ability to increase the structure of water.

Bromides↗

PubMed1978

A reexamination of the potential comedogenicity of sulfur.

The present study was designed to reexamine the question of the potential comedogenicity of topical sulfur. Studies were done by two different investigators in two separate institutions with groups of 12 and 40 subjects, respectively. Biopsy specimens were obtained from the occluded test sites before and after a six-week period of treatment. Each subject received a patch test of 5% sulfur in either an octoxynol 9 (Triton X-100) or a carbomer 934P (Carbopol 934P) vehicle and one of two controls consisting of the Triton X-100 vehicle or a dry patch test. We conclude that no trend or correlation was noted between the presence or absence of sulfur in the formulation and the appearance of comedones. The Triton X-100 vehicle itself appeared to be comedogenic under the experimental conditions.

Acne Vulgaris↗

PubMedDate not supplied

Direct analysis of platinum in plasma and urine by electrothermal atomic absorption spectrophotometry.

An improved technique is described for analysis of platinum (Pt) concentrations in plasma and urine by electrothermal atomic absorption spectrophotometry (EAAS). The method is intended for use in therapeutic monitoring of cancer patients treated with platinum-containing antitumor drugs. Samples (0.1 ml) of plasma, urine, or Pt standards are diluted to two ml with a matrix solution that contains diammonium edetate, ammonium dihydrogen phosphate, ammonium hydroxide, and octoxynol detergent. Concentrations of Pt in the diluted samples are determined directly by EAAS analysis with Zeeman background correction. Standard additions are unnecessary; Pt concentrations are read from a calibration chart of peak heights, which is linear up to 1.6 mg per liter. The detection limit is 0.02 mg of Pt per liter. Day-to-day precision (coefficient of variation, based on 21 consecutive runs) ranges from 4.2 to 11.7 percent, depending upon the Pt concentration in the plasma and urine specimens. Recovery of Pt added to plasma and urine specimens averages 103 +/- 8 and 99 +/- 6 percent, respectively. Concentrations of Pt are stable in plasma and urine specimens stored at 4 degrees C or -20 degrees C for four weeks. Analyses of Pt concentrations in serial plasma and urine specimens from cancer patients receiving cisplatin chemotherapy demonstrate the clinical utility of the technique.

Cisplatin↗

PubMed1987

Presence of abundant filaments in apical caps of the nonciliated bronchiolar epithelial (Clara) cells.

The nonionic detergent Triton X-100 has often been used for the extraction of cytoplasmic materials. We used the detergent in a vascular perfusion medium when preparing rat lung in order to observe the cytoskeleton of the nonciliated bronchiolar epithelial (Clara) cells. To eliminate some cytoplasmic materials selectively and to maintain good fine cell structure simultaneously, the lungs were perfused sequentially with the detergent (0.2% Triton X-100) alone for 2 min, with a mixture of low-concentration (0.1%) glutaraldehyde and detergent (0.2% Triton X-100) for 15 min, and finally with 2.5% glutaraldehyde for 5-10 min. After fixation, the nonciliated bronchiolar epithelial (Clara) cells were observed by scanning and transmission electron microscopy. At the apical region of the cells, there were central cytoplasmic protuberances (apical caps) filled with microfilaments. These filaments were bound at one end to the cytoplasmic side of the cell membrane and ran into the interior of the cytoplasm at the other end. As a control, the Clara cells were observed by transmission electron microscopy after perfusion with 2.5% glutaraldehyde solution. The luminal surfaces of the cells were covered with short, thick microvilli. The apical caps also had microvillus-like protrusions. These results suggest that the apical cap is not an apocrine droplet but rather is a stable structure involved in the function of the Clara cells.

Actin Cytoskeleton↗

PubMed1990

Functional, physical, and ultrastructural localization of CD15 antigens to the human polymorphonuclear leukocyte secondary granule.

A murine monoclonal IgM antibody, M3, which interferes with both polymorphonuclear leukocyte (PMN) phagocytosis and bactericidal activity, was used to examine the subcellular location of antigens bearing 3-fucosyllactosamine (CD15 antigens) within this cell type. Percoll gradient-separated secondary granule fractions were rich in CD15 antigens, with at least seven antigens recognizable in SDS-PAGE/electroblot studies. Sonication/sedimentation experiments using secondary granule fractions showed that both soluble and sedimentable CD15 antigens were present. Exposure of purified PMN to the secondary granule secretagogue phorbol myristate acetate caused extracellular release of two or three CD15 antigens, which could be purified by immunoprecipitation using antibody M3. Triton X-114 phase-partition experiments showed that secondary granule fraction CD15 antigens could be partitioned into hydrophilic (aqueous phase) and hydrophobic (detergent phase) antigens, suggesting that several of these antigens were integral secondary granule membrane components. Ultrastructurally, PMN intracellular granules showed two patterns of CD15 expression, localization over both granule matrix/granule membrane and localization to only granule membrane. Colocalization studies showed that lactoferrin and CD15 antigens were both present in a subset of intracellular granules, confirming a secondary granule location for these antigens.

Antibodies, Monoclonal↗

PubMed1991

A method for preparing skeletal muscle fiber basal laminae.

Previous attempts to prepare skeletal muscle basal laminae (BL) for ultrastructural analyses have been hampered by difficulties in successfully removing skeletal muscle proteins and cellular debris from BL tubes. In the present study we describe a two phase method which results in an acellular muscle preparation, the BL of which are examined by light, transmission electron, and scanning electron microscopy. In the first phase, excised rat extensor digitorum longus muscles are subjected to x-radiation and then soaked in Marcaine to inhibit muscle regeneration and to destroy peripheral muscle fibers. The muscles are then grafted back into their original sites and allowed to remain in place 7-14 days to allow for maximal removal of degenerating muscle tissue with minimal scar tissue formation. In the second phase, the muscle grafts are subjected sequentially to EDTA, triton X-100, DNAase, and sodium deoxycholate to remove phagocytizing cells and associated degenerating muscle tissue. These procedures result in translucent, acellular muscle grafts which show numerous empty tubes of BL backed by endomysial collagenous fibers. These preparations should be useful for morphological analyses of isolated muscle BL and for possible in vitro studies by which the biological activity of muscle BL can be examined.

Animals↗

PubMedDate not supplied

Release kinetics of ATP in cells exposed to nonionic detergents.

We have previously shown that the protein binding of intracellular ATP could be examined by monitoring the ATP release kinetics from Triton X-100 and Brij 58 nonionic detergent permeabilized cells. We have now analysed the protein binding of ATP in an isotonic medium using intact and partially ATP depleted Brij 58 treated human erythrocytes. The effects of Triton X-100 below the critical micelle concentration (CMC) was studied in normal and tumorous tissue culture cells and human red blood cells. Our results showed that the protein association of ATP was altered in the partially ATP depleted erythrocytes. Below the CMC value, but above a critical level Triton X-100 treatment was effective in mobilizing the intracellular ATP in both cell types. The ATP release curves were sigmoidal and an 'all or none' type of response was observed, especially in erythrocytes. The use of Triton X-100 (less than CMC) delays the detergent-induced cell decomposition time thus providing a new approach to investigating the physical state of intracellular ATP.

Adenosine Triphosphate↗

PubMed1989

Purification of the membrane protein enzyme lipoamidase by affinity chromatography.

Lipoamidase, a membrane glycoprotein enzyme, was purified from brain membrane by means of various affinity columns. A column with immobilized Arg-Phe-NH2 was found to be the most effective. After loading the crude material of the membrane, and extensive washing of the column with sodium chloride (0.3 M) solution, the enzyme activity was eluted by a solution containing 1% of nonionic detergent (Nonidet P-40). The fractions containing the lipoamidase activity were analyzed by SDS-PAGE, and a single protein band detected in this fraction. On the other hand, lipoyl-affinity columns with various resins were not effective in enzyme purification. Single step chromatography on the Arg-Phe-NH2 column enriched the membrane enzyme lipoamidase by 40-fold. The mechanism by which this affinity resin effectively enriches the enzyme remains to be elucidated.

Amidohydrolases↗

PubMed1985

Phospholipids in plant and animal chromatin.

Isolated hepatic nuclei and hepatic chromatin have been analysed for their DNA, RNA, protein and phospholipid content. The protein/DNA ratio is 3 for nuclei and 1.95 for chromatin extracted from Triton X-100 treated nuclei. The phospholipids, (2.36 +/- 0.91 (S.D.) per cent of the total nuclear material), are lost during the chromatin preparation mainly during the Triton X-100 washings of the nuclei. Nevertheless, 10 per cent of the total nuclear phospholipids remain bound to the chromatin. The comparative analysis of both nuclei and chromatin shows a difference in phospholipids and fatty acid composition. Thus, the chromatin-associated phospholipid cannot be attributed simply to contaminating nuclear membrane. This is supported by the autoradiographic study of semi-thin sections of interphase nuclei from root apices of Vicia faba in which [3H] ethanolamine is clearly localized in the chromatin and nucleolar regions of the nuclei.

Animals↗

PubMed1989

Clofibrate feeding increases cytoplasmic but not mitochondrial malic enzyme activity in rat kidney cortex.

Administration of clofibrate for 21 days to rats increased the malic enzyme activity in the kidney cortex by about 80 per cent. This effect seems to be specific since the drug did not alter significantly the activity either of lactate dehydrogenase, citrate synthase or total mitochondrial protein content in this organ. The increase in activity of malic enzyme in the 13,000 g supernatant (extramitochondrial) fraction in rats treated with the drug was about 80 per cent, whereas in the pellet (mitochondrial fraction) it was about 40 per cent. The specific activity of malic enzyme in the kidney cortex cytosol from clofibrate-treated rats was about twice that in controls. In contrast clofibrate treatment did not affect its specific activity in isolated mitochondria. Calculations showed that 0.57 and 0.53 mumoles min-1 g-1 wet tissue of mitochondrial malic enzyme was obtained in control and clofibrate-treated rats respectively. Thus, clofibrate feeding increases the amount of cytoplasmic but not mitochondrial malic enzyme activity.

Animals↗

PubMed1986

Regulation of sperm flagellar movement by protein phosphorylation and dephosphorylation.

Flagellar motility of Triton models of sea urchin spermatozoa was reactivated by cyclic AMP-dependent protein kinase and a protein factor, termed motility activator, both of which were prepared from the detergent-extract of sea urchin spermatozoa. It was shown that phosphorylation of the motility activator by the protein kinase is necessary for the reactivation of flagellar motility [Ishiguro et al, J. Cell Biol. 92:777-782, 1982; Murofushi et al, in "Biological Functions of Microtubules and Related Structures," Academic Press, 1982]. Reactivating factor was also detected in a KCl-extract of the axoneme fraction devoid of the detergent-extractable materials. The activity of this factor was also cyclic AMP- and protein kinase-dependent. Furthermore, when freshly prepared Triton models were treated with phosphoprotein phosphatase prepared from bovine cardiac muscle, the flagellar motility was drastically suppressed. This inhibition of the motility was partially recovered by the addition of cyclic AMP and protein kinase to the phosphatase-treated models.

Adenosine Triphosphatases↗

PubMed1986

Incorporation and turnover of labeled exogenous tubulin in the mitotic spindles of Chaetopterus oocytes and HeLa cells.

The incorporation of tubulin into mitotic spindles in situ was studied by incubating permeabilized mitotic cells in solutions containing [3H]GTP-labeled or dichlorotriazinylamino fluorescein (DTAF)-labeled tubulin. Metaphase HeLa cells or spindle-containing "minicells" from Chaetopterus oocytes were lysed in a microtubule-assembly buffer plus 0.5% Nonidet P-40, 1 mg/ml 120,000g supernatant mammalian brain tubulin, and [3H]GTP. After different periods of incubation, mitotic spindles were isolated in 2 M-glycerol-containing assembly buffer and separated from unbound counts by centrifugation through a 4 M-glycerol cushion; 3H counts per mg protein increase linearly for 8-12 min and then reach a plateau or steady state in both Chaetopterus oocytes and HeLa cells. Addition of 4 mM CaCl2 blocks or reverses incorporation. Little or no [3H]GTP is incorporated if exogenous tubulin or lysed cells are omitted from the assembly mixture. To measure the loss rate of [3H]GTP-tubulin from mitotic spindles, cells were incubated in tubulin plus [3H]GTP for 30 min, and a 20-fold excess of cold GTP (2 mM) was added. Samples were removed after incubation for different periods, and spindles were isolated as described above and counted for 3H content. [3H]GTP is lost from spindles at a rate of about 16%/min until a new steady state is reached in about 8 min. These results are consistent with an incorporation and turnover of [3H]GTP-tubulin in spindle microtubules of these lysed-cell models. The location of this newly incorporated tubulin in the spindle was investigated by incorporating fluorescent DTAF-tubulin into mitotic spindles of these lysed cell types.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Ultrastructure and motion analysis of permeabilized Paramecium capable of motility and regulation of motility.

Structural and behavioral features of intact and permeabilized Paramecium tetraurelia have been defined as a basis for study of Ca2+ control of ciliary reversal. Motion analysis of living paramecia shows that all the cells in a population swim forward with gently curving spirals at speeds averaging 369 +/- 19 microns/second. Ciliary reversal occurs in 10% of the cell population per second. Living paramecia, quick-fixed for scanning electron microscopy (SEM), show metachronal waves and an effective stroke obliquely toward the posterior end of the cell. Upon treatment with Triton X-100, swimming ceases and both scanning and transmission electron microscopy reveal cilia that uniformly project perpendicularly from the cell surface. Thin sections of these cells indicate that the ciliary, cell, and outer alveolar membranes are greatly disrupted or entirely missing and that the cytoplasm is also disrupted. These permeabilized paramecia can be reactivated and are capable of motility and regulation of motility. Motion analysis of cells reactivated with Mg2+ and ATP in low Ca2+ buffer (pCa greater than 7) shows that 71% swim forward in straight or curved paths at speeds averaging 221 +/- 20 microns/second. When these cells are quick-fixed for SEM the metachronal wave patterns of living, forward swimming cells reappear. Motion analysis of permeabilized cells reactivated in high Ca2+ buffers (pCa 5.5) shows that 94% swim backward in tight spirals at a velocity averaging 156 +/- 7 microns/second. SEM reveals a metachronal wave pattern with an effective stroke toward the anterior region. Although the permeabilized cells do not reverse spontaneously, the pCa response is preserved and the Ca2+ switch remains intact. The ciliary axonemes are largely exposed to the external environment. Therefore, the behavioral responses of these permeabilized cells depend on interaction of Ca2+ with molecules that remain bound to the axonemes throughout the extraction and reactivation procedures.

Animals↗