SEARCH · PubMed Health

Results for “Condensates”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 163 records · Page 9Linked to original sources

PubMed2001

Toxicology and carcinogenesis studies of coconut oil acid diethanolamine condensate (CAS No. 68603-42-9) in F344/N rats and B6C3F1 mice (dermal studies).

Coconut oil acid diethanolamine condensate, a mixture of fatty acid diethanolamides of the acids found in coconut oil, is widely used in cosmetics, shampoos, soaps, and related consumer products. Because of the lack of information about potential risks associated with long-term exposure, coconut oil acid diethanolamine condensate was selected as a representative of the diethanolamine chemical class for evaluation of toxicity and carcinogenic potential. Male and female F344/N rats and B6C3F1 mice received dermal applications of coconut oil acid diethanolamine condensate for 14 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, L5178Y mouse lymphoma cells, cultured Chinese hamster ovary cells, and mouse peripheral blood erythrocytes. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats received dermal applications of 0, 25, 50, 100, 200, or 400 mg coconut oil acid diethanolamine condensate/kg body weight in ethanol, five times per week for 14 weeks. All rats survived until the end of the study. Final mean body weights and body weight gains of 200 and 400 mg/kg males and females were significantly less than those of the vehicle controls. Clinical findings included irritation of the skin at the site of application in 100, 200, and 400 mg/kg males and females. Cholesterol concentrations were significantly decreased in 200 and 400 mg/kg males and in females administered 100 mg/kg or greater; triglyceride concentrations were also decreased in 200 and 400 mg/kg males. Histopathologic lesions of the skin at the site of application included epidermal hyperplasia, sebaceous gland hyperplasia, chronic active inflammation, parakeratosis, and ulcer. The incidences and severities of these skin lesions generally increased with increasing dose in males and females. The incidences of renal tubule regeneration in 100, 200, and 400 mg/kg females were significantly greater than the vehicle control incidence, and the severities in 200 and 400 mg/kg females were increased. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice received dermal applications of 0, 50, 100, 200, 400, or 800 mg coconut oil acid diethanolamine condensate/kg body weight in ethanol, five times per week for 14 weeks. All mice survived until the end of the study. Final mean body weights and body weight gains of dosed males and females were similar to those of the vehicle controls. The only treatment-related clinical finding was irritation of the skin at the site of application in males and females administered 800 mg/kg. Weights of the liver and kidney of 800 mg/kg males and females, the liver of 400 mg/kg females, and the lung of 800 mg/kg females were significantly increased compared to the vehicle controls. Epididymal spermatozoal concentration was significantly increased in 800 mg/kg males. Histopathologic lesions of the skin at the site of application included epidermal hyperplasia, sebaceous gland hyperplasia, chronic active inflammation, parakeratosis, and ulcer. The incidences and severities of these skin lesions generally increased with increasing dose in males and females. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats received dermal applications of 0, 50, or 100 mg coconut oil acid diethanolamine condensate/kg body weight in ethanol five times a week for 104 weeks. Survival, BODY WEIGHTS, AND CLINICAL FINDINGS: The survival rates of treated male and female rats were similar to those of the vehicle controls. The mean body weights of dosed males and females were similar to those of the vehicle controls throughout most of the study. The only chemical-related clinical finding was irritation of the skin at the site of application in 100 mg/kg females. PATHOLOGY FINDINGS: There were marginal increases in the incidences of renal tubule adenoma or carcinoma (combined) in 50 mg/kg females. The severity of nephropathy increased with increasing dose in female rats. Nonneoplastic lesions of the skin at the site of application included epidermal hyperplasia, sebaceous gland hyperplasia, parakeratosis, and hyperkeratosis, and the incidences and severities of these lesions increased with increasing dose. The incidences of chronic active inflammation, epithelial hyperplasia, and epithelial ulcer of the forestomach increased with dose in female rats, and the increases were significant in the 100 mg/kg group. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice received dermal applications of 0, 100, or 200 mg coconut oil acid diethanolamine condensate/kg body weight in ethanol five times a week for 104 to 105 weeks. SURVIVAL, BODY WEIGHTS, AND CLINICAL FINDINGS: Survival of dosed male and female mice was generally similar to that of the vehicle controls. Mean body weights of 100 mg/kg females from week 93 and 200 mg/kg females from week 77 were less than those of the vehicle controls. The only clinical finding attributed to treatment was irritation of the skin at the site of application in males administered 200 mg/kg. PATHOLOGY FINDINGS: The incidences of hepatic neoplasms (hepatocellular adenoma, hepatocellular carcinoma, and hepatoblastoma) were significantly increased in male and/or female mice. Most of the incidences exceeded the historical control ranges. The incidences of eosinophilic foci in dosed groups of male mice were increased relative to that in the vehicle controls. The incidences of renal tubule adenoma and renal tubule adenoma or carcinoma (combined) were significantly increased in 200 mg/kg males. Several nonneoplastic lesions of the skin at the site of application were considered treatment related. Incidences of epidermal hyperplasia, sebaceous gland hyperplasia, and hyperkeratosis were greater in all dosed groups of males and females than in the vehicle controls. The incidences of ulcer in 200 mg/kg males and inflammation and parakeratosis in 200 mg/kg females were greater than those in the vehicle controls. The incidences of thyroid gland follicular cell hyperplasia in all dosed groups of males and females were significantly greater than those in the vehicle control groups. GENETIC TOXICOLOGY: Coconut oil acid diethanolamine condensate did not show genotoxic activity in vitro. It was not mutagenic in Salmonella typhimurium, nor did it produce an increase in mutant L5178Y mouse lymphoma cell colonies. In addition, no increases in the frequencies of sister chromatid exchanges or chromosomal aberrations were observed in Chinese hamster ovary cells after incubation with coconut oil acid diethanolamine condensate. All these in vitro assays were conducted with and without induced S9 activation enzymes. In contrast to the uniformly negative results in vitro, positive results were obtained in a peripheral blood micronucleus test in male and female mice from the 14-week dermal study. CONCLUSIONS: Under the conditions of these 2-year dermal studies, there was no evidence of carcinogenic activity of coconut oil acid diethanolamine condensate in male F344/N rats administered 50 or 100 mg/kg. There was equivocal evidence of carcinogenic activity in female F344/N rats based on a marginal increase in the incidences of renal tubule neoplasms. There was clear evidence of carcinogenic activity in male B6C3F1 mice based on increased incidences of hepatic and renal tubule neoplasms and in female B6C3F1 mice based on increased incidences of hepatic neoplasms. These increases were associated with the concentration of free diethanolamine present as a contaminant in the diethanolamine condensate. Exposure of rats to coconut oil acid diethanolamine condensate by dermal application in ethanol for 2 years resulted in epidermal hyperplasia, sebaceous gland hyperplasia, hyperkeratosis, and parakeratosis in males and females and ulcer in females at the site of application. There were increases in the incidences of chronic inflammation, epithelial hyperplasia, and epithelial ulcer in the forestomach of female rats. The severities of nephropathy in dosed female rats were increased. Exposure of mice to coconut oil acid diethanolamine condensate by dermal application for 2 years resulted in increased incidences of eosinophilic foci of the liver in males. Increased incidences of epidermal hyperplasia, sebaceous gland hyperplasia, and hyperkeratosis in males and females, ulcer in males, and parakeratosis and inflammation in females at the site of application and of follicular cell hyperplasia in the thyroid gland of males and females were chemical related.

Administration, Topical↗

PubMed1995

Relationships between cellular condensation, preosteoblast formation and epithelial-mesenchymal interactions in initiation of osteogenesis.

Initiation of osteogenesis or bone formation is dependent on cell and tissue interactions. We investigated the events between 4 and 7 days of incubation that translate epithelial-mesenchymal signalling into overt differentiation of osteoblasts and deposition of bone in the mandibles of chick embryos. Condensation of mandibular mesenchyme (the membranous skeleton), visualized with PNA-lectin, occurred at H.H. mid-26 (5.75 days), lasted 12 h and preceded osteoblast differentiation by 1.5 days. As determined from 3D-reconstruction all mandibular membrane bones arose from a single condensation closely associated with the stomodeal epithelium. The finding that the osteogenic condensation in the mandibular arch is a major branch of a common condensation that provides osteogenic mesenchyme to both maxillary and mandibular arches establishes a closer link between mechanisms controlling development of the skeleton in these two arches than previously suspected. Preosteoblasts (alkaline phosphatase-positive cells) form in the mandible at H.H. early 25, which is before condensation but after the epithelial-mesenchymal interaction upon which preosteoblast formation and condensation depend--neither form in isolated mesenchyme, whereas both form after recombination of mesenchyme and epithelium. Tenascin was present in the mandibular epithelium only at H.H. stage 19 but not in the mesenchyme at any age. Therefore, the epithelial-mesenchymal interaction controls initiation of osteogenesis at the preosteoblast stage. Preosteoblasts then condense, transform into osteoblasts and deposit bone matrix. Differentiation of preosteoblasts precedes condensation which amplifies their number. This is in contrast with chondrogenesis where condensation triggers prechondroblast differentiation.

Animals↗

PubMedDate not supplied

Stage-specific onset of condensation and matrix deposition for Meckel's and other first arch cartilages in inbred C57BL/6 mice.

Condensation is a multistep process, involving cell recruitment, cell-to-cell contact by cell adhesions, and concomitant changes in cell shape. Condensation of prechondrogenic cells down-regulates and/or inactivates cell proliferation and enhances the activities of cartilage-specific genes. Timing of onset and duration of condensation are thus important regulatory processes mediating cellular and molecular events during chondrogenesis. The present study was undertaken to examine timing of onset and duration of condensation and onset of matrix formation for first arch cartilages in inbred C57BL/6 mice. Because timing can only be reliably assessed in very precisely staged embryos, mice were mated for only 2 hr, pregnancy was determined by weight increase, and embryos were assigned to substages and divisions of Theiler's [1972] stages on the basis of external development of the frontonasal area, eyes, vibrissae, mandibular and hyoid auricular hillocks (pinna) and some internal structures [Miyake et al., 1996]. Condensation and matrix formation were determined using PNA lectin histochemistry, type II collagen immunohistochemistry, Mallory's trichrome, Hall and Brunt's quadruple and toluidine blue stains on serially sectioned embryos, and 3-D reconstruction. A single, continuous, first arch chondrogenic condensation was identified. It consisted of three components: 1) a rostral component for the symphysis; 2) a core component for the major portion of Meckel's cartilage; and 3) a caudal component for the caudo-lateral area of Meckel's cartilage and the two ear cartilages, the malleus and incus. The caudal component was further divided into rostral and caudal subcomponents which gave rise to the malleus and incus, respectively. Cellular arrangement differed among the three components, probably reflecting major axes of matrix growth. The condensation was only weakly recognized by peanut agglutinin lectin (PNA), unless sections were pretreated with neuraminidase, showing the masking effects of sialic acid. The core and caudal components of the condensation appeared at stage 20.12 and ended at stage 21.14 and 21.32, respectively. The symphysial component began at stage 20.2 and ended at stage 21.32. Deposition of sulfated proteoglycans began at stage 21.14 in the core component of the condensation--preceding slightly deposition of type II collagen--and at stage 21.32 in the symphysial and caudal components.

Animals↗

PubMedDate not supplied

Effect of condensation techniques on amalgam bond strengths to dentin.

This in vitro study determined the shear bond strengths of Tytin amalgam to dentin using All-Bond 2, Amalgambond Plus, Amalgambond Plus with HPA powder, and OptiBond with hand and mechanical (Condensaire) condensation. The occlusal enamel surfaces were ground flat to expose the dentin surfaces, and polished with 600-grit SiC paper. The dentin surfaces were treated with one of the combinations of a dentin bonding agent and a condensation technique. All specimens were thermocycled 300 cycles in water between 5-55 degrees C, and shear bond strengths were determined with a Zwick Universal Testing Machine. Two-way ANOVA and Duncan's tests showed no significant differences (P > 0.05) in shear bond strength between hand and mechanical condensation of the paired groups, with the exception of Amalgambond Plus, where hand condensation was shown to be significantly better. For both condensation techniques, shear bond strength of Amalgambond Plus with HPA powder was significantly higher than the other materials. For mechanical condensation, OptiBond was significantly higher than either Amalgambond Plus or All-Bond 2, and no significant difference was found between Amalgambond Plus and All-Bond 2. For hand condensation, OptiBond was only significantly higher than All-Bond 2, while there was no significant difference between OptiBond and Amalgambond Plus or between Amalgambond Plus and All-Bond 2. In conclusion, for both condensation techniques, Amalgambond Plus with HPA powder performed the best in bonding amalgam to dentin. The two different condensation techniques did not significantly affect shear bond strength, except with Amalgambond Plus.

Analysis of Variance↗

PubMed1998

An indirect role for cyclin B-Cdc2 in inducing chromosome condensation in Xenopus egg extracts.

We have studied the cytoplasmic mechanism that induces metaphase chromosome condensation in cell-free Xenopus egg extracts. To analyze the mechanism responsible for inducing chromosome condensation separately from those responsible for sperm chromatin remodeling and nuclear envelope disassembly, we used Xenopus sperm chromatin that had already been remodeled to nucleosomal chromatin by incubating demembranated sperm with egg extracts added with lysolecithin. We found that inhibition of cyclin B-Cdc2 with butyrolactone I abolished chromosome condensation of the remodeled sperm chromatin by M-phase egg extracts, but incubation of the chromatin with active cyclin B-Cdc2 alone did not induce chromosome condensation, indicating a requirement for cytoplasmic factor(s) in addition to cyclin B-Cdc2 for the induction of chromosome condensation. We further demonstrated that if the cyclin B-Cdc2-dependent phosphorylation state was protected against dephosphorylation by a preincubation of M-phase extracts with ATP-gamma-S, chromosome condensation and phosphorylation of chromosomal histone H1 occurred even when extracts were depleted of cyclin B-Cdc2 activity. The chromosome condensation seen in the absence of cyclin B-Cdc2 was completely inhibited with another protein kinase inhibitor, 6-dimethylaminopurine, implying that a protein kinase other than cyclin B-Cdc2 was involved in the induction of chromosome condensation. These results strongly suggest that a cyclin B-Cdc2-dependent protein kinase cascade is involved in inducing chromosome condensation and the phosphorylation of chromosomal histone H1.

Animals↗

PubMed2025

An RNA Condensate Model for the Origin of Life.

The RNA World hypothesis predicts that self-replicating RNAs evolved before DNA genomes and coded proteins. Despite widespread support for the RNA World, self-replicating RNAs have yet to be identified in a natural context, leaving a key 'missing link' for this explanation of the origin of life. Inspired by recent work showing that condensates of charged polymers are capable of catalyzing chemical reactions, we consider a catalytic RNA condensate as a candidate for the self-replicating RNA. Specifically, we propose that short, low-complexity RNA polymers formed catalytic condensates capable of templated RNA polymerization. Because the condensate properties depend on the RNA sequences, RNAs that formed condensates with improved polymerization and demixing capacity would be amplified, leading to a 'condensate chain reaction' and evolution by natural selection. Many of the needed properties of this self-replicating RNA condensate have been realized experimentally in recent studies and our predictions could be tested with current experimental and theoretical tools. Our theory addresses central problems in the origins of life: (i) the origin of compartmentalization, (ii) the error threshold for the accuracy of templated replication, (iii) the free energy cost of maintaining an information-rich population of replicating RNA polymers. Furthermore, we note that the extant nucleolus appears to satisfy many of the requirements of an evolutionary relic for the model we propose. More generally, we suggest that future work on the origin of life would benefit from condensate-centric biophysical models of RNA evolution.

Origin of Life↗

PubMed2000

All for one and one for all: condensations and the initiation of skeletal development.

Condensation is the pivotal stage in the development of skeletal and other mesenchymal tissues. It occurs when a previously dispersed population of cells gathers together to differentiate into a single cell/tissue type such as cartilage, bone, muscle, tendon, kidney, and lung and is the earliest stage during organ formation when tissue-specific genes are upregulated. We present a synopsis of our current understanding of how condensations are initiated and grown, how their boundaries and sizes are set, how condensation ceases, and how overt differentiation begins. Extracellular matrix molecules, cell surface receptors and cell adhesion molecules, such as fibronectin, tenascin, syndecan, and N-CAM, initiate condensation formation and set condensation boundaries. Hox genes (Hoxd-11-13) and other transcription factors (CFKH-1, MFH-1, osf-2), modulate the proliferation of cells within condensations. Cell adhesion is ensured indirectly through Hox genes (Hoxa-2, Hoxd-13), and directly via cell adhesion molecules (N-CAM and N-cadherin). Subsequent growth of condensations is regulated by BMPs, which activate Pax-2, Hoxa-2 and Hoxd-11 among other genes. Growth of a condensation ceases when Noggin inhibits BMP signalling, setting the stage for transition to the next stage of skeletal development, namely overt cell differentiation. BioEssays 22:138-147, 2000.

Animals↗

PubMedDate not supplied

Vibrational CD (VCD) and atomic force microscopy (AFM) study of DNA interaction with Cr3+ ions: VCD and AFM evidence of DNA condensation.

The interaction of natural calf thymus DNA with Cr(3+) ions was studied at room temperature by means of vibrational CD (VCD) and infrared absorption (ir) spectroscopy, and atomic force microscopy (AFM). Cr(3+) ion binding mainly to N(7) (G) and to phosphate groups was demonstrated. Psi-type VCD spectra resembling electronic CD (ECD) spectra, which appear during psi-type DNA condensation, were observed. These spectra are characterized mainly by an anomalous, severalfold increase of VCD intensity. Such anomalous VCD spectra were assigned to DNA condensation with formation of large and dense particles of a size comparable to the wavelength of the probing ir beam and possessing large-scale helicity. Atomic force microscopy confirmed DNA condensation by Cr(3+) ions and the formation of tight DNA particles responsible for the psi-type VCD spectra. Upon increasing the Cr(3+) ion concentration the shape of the condensates changed from loose flower-like structures to highly packed dense spheres. No DNA denaturation was seen even at the highest concentration of Cr(3+) ions studied. The secondary structure of DNA remained in a B-form before and after the condensation. VCD and ir as well as AFM proved to be an effective combination for investigating DNA condensation. In addition to the ability of VCD to determine DNA condensation, VCD and ir can in the same experiment provide unambiguous information about the secondary structure of DNA contained in the condensed particles.

Animals↗

PubMed1990

Approach to the limit of counterion condensation.

According to counterion condensation theory, one of the contributions to the polyelectrolyte free energy is a pairwise sum of Debye-Hückel potentials between polymer charges that are reduced by condensed counterions. When the polyion model is taken as an infinitely long and uniformly spaced line of charges, a simple closed expression for the summation, combined with entropy-derived mixing contributions, leads to the central result of the theory, a condensed fraction of counterions dependent only on the linear charge density of the polyion and the valence of the counterion, stable against increases of salt up to concentrations in excess of 0.1 M. Here we evaluate the sum numerically for B-DNA models other than the infinite line of B-DNA charges. For a finite-length line there are end effects at low salt. The condensation limit is reached as a flat plateau by increasing the salt concentration. At a fixed salt concentration the condensation limit is reached by increasing the length of the line. At moderate salt even very short B-DNA line-model oligomers have condensed fractions not far from the infinite polymer limit. For a long double-helical array with charge coordinates at the phosphates of B-DNA, the limiting condensed fraction appears to be approached at low salt. In contrast to the results for the line of charges, however, the computed condensed fraction varies strongly with salt in the range of experimentally typical concentrations. Salt invariance is restored, in agreement with both the line model and experimental data, when dielectric saturation is considered by means of a distance-dependent dielectric function. For sufficiently long B-DNA line and helical models, as typical salt concentrations, the counterion binding fraction approaches the polymer limit as a linear function of 1/P, where P is the number of phosphate groups of B-DNA.

Animals↗

PubMed1988

Condensation of DNA in situ in metaphase chromosomes induced by intercalating ligands and its relationship to chromosome banding.

Interactions of certain intercalating cationic ligands with nucleic acids result in the formation of products that undergo condensation and agglomeration; this transition in solution can be monitored by light-scatter measurements. In the present study, using such intercalators as the antitumor drug mitoxantrone or fluorochromes acridine orange and quinacrine, we induced condensation of DNA in situ in Chinese hamster chromosomes. The in situ products scattered light and could be detected by darkfield- or phase-contrast microscopy. In the darkfield the complexes had a characteristic granular appearance and often generated a banding pattern on the chromosomes. In contrast, condensation of DNA in situ by the nonintercalating polyvalent cations (Co3+, spermine4+), while enhancing the chromosome's image contrast, did not produce the granular products or the banding. The condensation of free DNA, single or double stranded, natural or synthetic, the latter of various base composition and configuration, was also measured in solution. The condensation in solution and in situ was observed at similar concentrations of the respective ligands. The intercalating dye ethidium bromide, which did not condense DNA in solutions of moderate and high ionic strength, also did not generate the granular products or banding on chromosomes. The data also show that both base composition and configuration are important factors in determining the sensitivity of DNA to condensation by particular intercalating ligands. The studies suggest that the phenomenon of DNA condensation by intercalating dyes, which shows a high degree of specificity with respect to primary and secondary structures of DNA, may be associated with mechanisms of chromosome banding induced by the intercalating thiazine dyes in Giemsa staining or by quinacrine. Observation of chromosome banding based on light-scatter detection in darkfield microscopy allows the study of interactions between DNA and the ligands that neither fluoresce nor generate colored products. This principle of chromosome "counter-staining" can be explored by flow cytometry.

Acridine Orange↗

PubMed1991

Propionyl-CoA condensing enzyme from Ascaris muscle mitochondria. I. Isolation and characterization of multiple forms.

The condensation of two propionyl-CoA units or a propionyl-CoA with acetyl-CoA is required for the synthesis of 2-methylvalerate or 2-methylbutyrate, respectively, two of the major fermentation products of Ascaris anaerobic muscle metabolism. An enzyme that preferentially catalyzes the condensation of propionyl-CoA rather than acetyl-CoA has been purified from the mitochondria of the parasitic intestinal nematode Ascaris lumbricoides var. suum. The purified enzyme is over 10 times more active with propionyl-CoA than with acetyl-CoA as substrate. It also catalyzes the coenzyme A-dependent hydrolysis of acetoacetyl-CoA at a rate four times higher than the propionyl-CoA condensation reaction. The purified Ascaris condensing enzyme preferentially forms the 2-methyl-branched-chain keto acids rather than the corresponding straight chain compounds. The native molecular weight of the purified enzyme was estimated to be 160,000 by gel filtration chromatography and 158,000 by high pressure liquid chromatography. The enzyme migrated as a single protein band with Mr 40,000 during sodium dodecyl sulfate-polyacrylamide electrophoresis, indicating that the enzyme is composed of four subunits of the same molecular weight. Chromatography on CM-sephadex resulted in the isolation of two separate peaks of activity, designated as A and B. Both A and B had the same molecular weight and subunit composition. However, they differed in their specific activities and isoelectric points. The pIs of condensing enzymes A and B were 7.6 and 8.4, respectively. Propionyl-CoA was the best substrate for the condensation reaction with both enzymes. However, the specific activity of enzyme B for both propionyl-CoA condensation (3.4 mumol/min/mg protein) and acetoacetyl-CoA thiolysis (13.8 mumol/min/mg protein) was 2.4 times higher than that obtained with enzyme A. Similarly, chromatography on phosphocellulose resolved the Ascaris condensing enzyme activity into one minor and two major peaks. All of these components had the same molecular weight and subunit composition, but differed in their specific activities. The two major phosphocellulose peaks cross-reacted immunologically when examined by the Ouchterlony double immunodiffusion technique. In addition, antiserum against the phosphocellulose most active form cross-reacted with forms A and B isolated by chromatography of the enzyme on CM-Sephadex, indicating that all forms were immunochemically related.

Alcohol Oxidoreductases↗

PubMed2003

Sequence-dependent DNA condensation and the electrostatic zipper.

Sequence-dependent configuration changes and condensation of double-stranded poly(dG-dC).(dG-dC) (GC-DNA) and ds poly(dA-dT).(dA-dT) (AT-DNA) were observed by atomic force microscopy in the presence of Ni(II). Less condensing agent was required to generate configuration changes in GC-DNA as compared to AT-DNA. In the presence of Ni(II) cations, GC-DNA adopted a Z-type conformation and underwent a stepwise condensation, starting with partial intramolecular folding, followed by intermolecular condensation of two to several molecules and ending with the formation of toroids, rods, and jumbles. GC-DNA condensates were unusual in that the most highly condensed regions were surrounded by loops of ds GC-DNA. In contrast, AT-DNA retained its B-type conformation and displayed only minor condensation even at high Ni(II) concentrations. The Ni(II)-dependent differences in condensation between GC-DNA and AT-DNA are predicted by an extension of the electrostatic zipper motif proposed by Kornyshev and Leikin, in which we account for shorter than Debye screening length surface separations between the DNA molecules and for the Ni(II)-induced conformation change of GC-DNA to Z-DNA.

DNA↗

PubMed1998

Insertion of telomere repeat sequence decreases plasmid DNA condensation by cobalt (III) hexaammine.

Telomere repeat sequence (TRS) DNA is found at the termini of most eukaryotic chromosomes. The sequences are highly repetitive and G-rich (e.g., [C(1-3)A/TG(1-3)]n for the yeast Saccharomyces cerevisiae) and are packaged into nonnucleosomal protein-DNA structures in vivo. We have used total intensity light scattering and electron microscopy to monitor the effects of yeast TRS inserts on in vitro DNA condensation by cobalt (III) hexaammine. Insertion of 72 bp of TRS into a 3.3-kb plasmid depresses condensation as seen by light scattering and results in a 22% decrease in condensate thickness as measured by electron microscopy. Analysis of toroidal condensate dimensions suggests that the growth stages of condensation are inhibited by the presence of a TRS insert. The depression in total light scattering intensity is greater when the plasmid is linearized with the TRS at an end (39-49%) than when linearized with the TRS in the interior (18-22%). Circular dichroism of a 95-bp fragment containing the TRS insert gives a spectrum that is intermediate between the A-form and B-form, and the anomalous condensation behavior of the TRS suggests a noncanonical DNA structure. We speculate that under conditions in which the plasmid DNA condenses, the telomeric insert assumes a helical geometry that is similar to the A-form and is incompatible with packing into the otherwise B-form lattice of the condensate interior.

Base Sequence↗

PubMed1990

Ultrasonic condensation of gutta-percha: an in vitro dye penetration and scanning electron microscopic study.

Lateral condensation of gutta-percha with and without ultrasonic activation of the spreader was compared by use of dye penetration analysis and scanning electron photomicrographs of the gutta-percha fills in extracted human incisors and canines. The root canal fillings in three groups of 10 teeth each were laterally condensed using an ultrasonically activated spreader, a fine finger spreader, or a non-activated ultrasonic spreader. Significantly less apical dye penetration occurred when teeth were obturated using an ultrasonically activated spreader as compared with manual condensation with fine finger spreaders. The ultrasonically condensed gutta-percha mass was more homogeneous with fewer voids compared with gutta-percha masses from the two groups that were condensed without ultrasonic activation. A correlation between apical microleakage and the appearance of the gutta-percha mass was seen. All teeth with well-condensed, more homogeneous fillings had low dye penetration. All teeth with a high extent of dye penetration had poorly condensed, less homogeneous fillings. However, not all teeth that had poorly condensed fillings had high dye penetration.

Cuspid↗

PubMed2000

De-misting condensation on intraocular lenses.

BACKGROUND: Condensation occurs rapidly on intraocular lenses (IOLs), particularly silicone IOLs, after vitrectomy and fluid-air exchange in the presence of a posterior capsulotomy and severely limits the surgeon's view of the retina. This study investigated the value of heating contact lens irrigation fluid to prevent condensation on IOLs. DESIGN: An experimental in vitro study and a noncomparative interventional case series. PARTICIPANTS: Five control eyes had temperature measurements during fluid-air exchanges. Two patients with silicone IOLs and posterior capsulotomies underwent a vitrectomy involving a fluid-air exchange with heated contact lens fluid. METHODS: An in vitro model allowed monitoring of temperature and humidity changes during condensation formation on four different IOL materials. Adjusting variables to promote evaporation rather than condensation was achieved in vitro. In vivo, intraocular temperatures were measured at various stages of five vitrectomies involving a fluid-air exchange. Finally, in two in vivo cases a blood warmer was used to heat the accessory contact lens irrigation fluid during the vitrectomy and fluid-air exchange. MAIN OUTCOME MEASURES: Successful prevention of condensation on the silicone IOL during the fluid-air exchange. RESULTS: Anterior segment temperature influences the IOL temperature, such that when it is higher than the posterior segment temperature, condensation does not form and evaporation is promoted. In vivo the temperatures in the eye are hypothermic throughout the vitrectomy. At fluid-air exchange the posterior segment heats rapidly as irrigation ceases and low specific heat gas enters. Finally, in two in vivo cases with silicone IOLs, condensation was predicted then prevented by our intervention for the duration of the fluid-air exchange. CONCLUSIONS: Heating the anterior segment by conduction from warmed irrigation fluid flowing through the irrigating contact lens represents a cheap, noninvasive, and safe means to prevent condensation on IOL materials.

Body Temperature↗

PubMed1998

In vitro condensation-sorting of enzyme proteins isolated from rat pancreatic acinar cells.

To study the process of granule formation in pancreatic acinar cells in more detail we have established an in vitro system in which the whole complement of enzyme proteins released from isolated zymogen granules is mixed with a tracer amount of the same biosynthetically labeled proteins and is incubated at conditions prevailing in either pre-Golgi (pH 7.5) or trans-Golgi (pH 5.9) compartments. Condensation of the proteins into dense cores is assayed and quantitated after centrifugation of the mixture at 13000g and separation of the proteins in both the supernatant and the pellet by 2D-gel electrophoresis. At pH 7.5 about 1% of the total protein-bound radioactivity can be sedimented into the pellet and this increases 5-fold at pH 5.9 with similar sedimentation efficiency for individual enzyme proteins. The usual assumption that all aggregated proteins can be sedimented and thus only the pellet is representative for pH-dependent condensation has to be modified by the fine structural analysis of both the supernatant and pellet fraction at pH 7.5 and 5.9. Small particulate complexes form already in the supernatant at pH 7.5 which are not sedimented to a large extent into the pellet. At pH 5.9 aggregates of a homogeneous size of about 0.6 to 0.8 microm formed in the supernatant while the pellet is composed of sheets and vesicles of membranes studded with dense core particles of about 20 to 30nm size. The pH-dependent protein condensation is a stepwise process starting with the formation of small dense core particles already at pH 8.0/7.5 which then progressively aggregate to form larger cores at pH 6.0/5.0. These aggregates can only be sedimented employing higher centrifugal forces. In the condensation process of pancreatic enzyme proteins calcium ions exert an effect only at pH 7.5, leading to somewhat larger dense particles, while potassium ions are inhibitory both in protein condensation and in the binding of particles to membranes. The process of pH-dependent protein condensation is reversible and can be performed repetitively. The sedimentation of condensed proteins can be increased by the addition of isolated zymogen granule membranes. Thus the in vitro system allows the analysis of two related processes in granule formation: the condensation of secretory proteins into granule cores and their binding to the granule membrane.

Animals↗

PubMed2000

Strategies for maintaining the particle size of peptide DNA condensates following freeze-drying.

The particle size of peptide DNA condensates were studied after freeze-drying and rehydration as a function of sugar excipient, concentration, pH, DNA concentration, and peptide condensing agent. In the absence of an excipient, freeze-dried 50 microg/ml AlkCWK(18) (iodoacetic acid alkylated Cys-Typ-Lys(18)) DNA condensates formed large fibrous flocculates on rehydration. Of the sugars tested as lyoprotectants, sucrose proved most effective at preserving particle size during rehydration. The addition of 5 wt/vol% sucrose preserved a mean particle diameter of less than 50 nm during rehydration of AlkCWK(18) DNA condensates prepared at DNA concentrations up to 200 microg/ml; however, higher DNA concentrations led to the formation of insoluble fibrous flocculates. Substitution of polyethylene glycol (PEG)-CWK(18) as a DNA condensing peptide eliminated the need for sucrose, resulting in peptide DNA condensates that retained particle size when rehydrated in water or normal saline at concentrations up to 5 mg/ml. The results suggest that sucrose functions primarily as a bulking agent during freeze-drying that only preserves the particle size of AlkCWK(18) DNA condensates up to a maximum concentration of 200 microg/ml. Alternatively, the steric layer created on the surface of PEG-CWK(18) DNA condensates provides far more efficient lyoprotection, preserving their particle size at a concentration of 5 mg/ml without a bulking agent.

DNA↗

PubMed1995

Condensation of plasmids enhanced by Z-DNA conformation of d(CG)n inserts.

DNA molecules collapse into compact structures in the presence of multivalent cations. To probe the possible importance of supercoiling and conformational effects, pUC18 plasmids (2686 bp) were modified by inserting 12-bp and 20-bp alternating d(CG)n sequences, which are capable of converting to a left-handed Z-conformation under appropriate conditions, into the polycloning region. Condensation was induced by rapid addition of hexaammine cobalt(III) [Co(NH3)6(3+)] and monitored by laser light scattering and electron microscopy. Light scattering shows that plasmids with longer d(CG)n inserts condense more extensively at natural superhelical densities. Electron microscopy indicates that the morphological distribution of condensed d(CG)n-containing plasmids changes as a function of Co(NH3)6(3+) concentration. At lower Co(NH3)6(3+) concentration, the proportion of rods is higher, and at higher Co-(NH3)6(3+) concentration, most of the condensates have the form of toroids. In addition, the inner radii of the toroids are much smaller relative to condensed pUC18 under the same conditions. Enzymatic analysis and chemical probing show that the d(CG)n inserts in naturally supercoiled plasmids have extensively converted from B-form to Z-form in the presence of Co(NH3)6(3+) at the upper range of concentrations under which condensation occurs. To determine whether the enhanced condensation of d(CG)n-containing plasmids results from the change of superhelical density due to the B-Z transition, we treated wild-type pUC18 molecules with topoisomerase I and varying amounts of ethidium bromide to generate a range of supercoil densities. Light scattering indicates that supercoiling did not affect the condensation process.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗