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Increased resistance of hygroscopic condenser humidifiers when using a closed circuit suction system.

OBJECTIVES: To examine a hygroscopic condenser after clinical use and to describe the interaction of a hygroscopic condenser and a closed circuit suction system used simultaneously. DESIGN: Prospective evaluation of hygroscopic condensers used clinically, and laboratory investigation of a hygroscopic condenser used with a closed circuit suction system. SETTING: Tertiary referral centers. MEASUREMENTS AND MAIN RESULTS: The hygroscopic condenser used during mechanical ventilation was removed and peak inflation pressure was measured by delivering a standard tidal volume and inspiratory flow across the isolated hygroscopic condenser while recording the peak inflation pressure. In the laboratory, four 10-mL aliquots of saline were instilled via closed circuit suction system into a test lung with fresh hygroscopic condensers (n = 15) inline. At baseline and after each instillation, the hygroscopic condenser was weighed and the peak inflation pressure was measured while in five condensers, peak expiratory flow rate was also measured. In these five devices, hygroscopic condenser resistance was measured with 100 L/min of constant gas flow while measuring the pressure drop across the hygroscopic condenser. In 11 hygroscopic condensers used for 27.5 +/- 11.9 hrs with no closed circuit suction system, the peak inflation pressure was 3.74 +/- 0.58 cm H2O. In the laboratory, instillation of saline via closed circuit suction system was associated with an increase in hygroscopic condenser weight. Peak inflation pressure increased in a quadratic fashion with the increase in hygroscopic condenser weight, while peak expiratory flow rate decreased in a linear fashion. After four saline instillations, hygroscopic condenser resistance increased from 5.66 +/- 0.31 to 13.9 +/- 2.42 cm H2O/L/sec. CONCLUSIONS: Clinical use of a hygroscopic condenser alone is not associated with a significant increase in peak inflation pressure. We caution the use of a hygroscopic condenser and a closed circuit suction system simultaneously, as an increase in hygroscopic condenser resistance may develop and may be poorly tolerated in patients with marginal ventilatory reserve.

Aged↗

Mutagenicity of wood smoke condensates in the Salmonella/microsome assay.

Smoke condensates of woods used for food preservation and aromatization in Nigeria were tested for mutagenic activity using Salmonella typhimurium TA98 and TA100. The woods were: white mangrove (Avicennia nitida), red mangrove (Rhizophora racemosa), mahogany Khaya sp.), abura (Mitragyna ciliata), alstonia (Alstonia boonei) and black afara (Terminalia ivorensis). Cigarette tar was tested for comparison. The condensates induced dose-dependent increases in the number of His+ revertants mainly with S9 mix. With the exception of mahogany and cigarette smoke condensate, the smoke condensates induced more revertants/microgram condensate in TA100 than in TA98. The number of revertants/microgram condensate ranged between 0.04 and 0.9 for the wood smoke condensates and was 0.12 for the cigarette smoke in TA100. The range was between 0.1 and 0.30 for the wood smoke condensates and 0.18 revertants/microgram condensate for cigarette smoke condensate in TA98. Concentrations of 7 polycyclic aromatic hydrocarbons (PAHs) in the condensates were determined namely, pyrene, benzo[a]pyrene, benz[a]anthracene, benzo[k]fluoranthene, benzo[b]chrysene, benzo[g,h,i]perylene and dibenzo[a,e]pyrene. The condensates contained varying concentrations of the individual PAHs and those with higher concentrations generally showed greater mutagenic activities. However, the order of mutagenic potency in the bacterial strains differed from the order of PAH concentrations, which were lower than the concentrations at which they are reported to induce mutations. When 6 of the PAHs were mixed in the concentrations in which they were found in the individual condensates, the mixtures did not induce mutation so that the contribution of the PAHs to the mutagenic activities of the condensates could not be determined.

Microsomes↗

The membranous skeleton: the role of cell condensations in vertebrate skeletogenesis.

Elements of the vertebrate skeleton are initiated as cell condensations, collectively termed the 'membranous skeleton' whether cartilages or bones by Grüneberg (1963). Condensations, which were identified as the basic cellular units in a recent model of morphological change in development and evolution (Atchley and Hall 1991) are reviewed in this paper. Condensations are initiated either by increased mitotic activity or by aggregation of cells towards a centre. Prechondrogenic (limb bud) and preosteogenic (scleral ossicle) condensations are discussed and contrasted. Both types of skeletogenic condensations arise following epithelial-mesenchymal interactions; condensations are identified as the first cellular product of such tissue interactions. Molecular characteristics of condensations are discussed, including peanut agglutinin lectin, which is used to visualize prechondrogenic condensations, and hyaluronan, hyaladherins, heparan sulphate proteoglycan, chondroitin sulphate proteoglycan, versican, tenascin, syndecan, N-CAM, alkaline phosphatase, retinoic acid and homeo-box-containing genes. The importance for the initiation of chondrogenesis or osteogenesis of upper and lower limits to condensation size and the numbers of cells in a condensation are discussed, as illustrated by in vitro studies and by mutant embryos, including Talpid3 in the chick and Brachypod, Congenital hydrocephalus and Phocomelia in the mouse. Evidence that genes specific to the skeletal type are selectively activated at condensation is discussed, as is a recent model involving TGF-beta and fibronectin in condensation formation. Condensations emerge as a pivotal stage in initiation of the vertebrate skeleton in embryonic development and in the modification of skeletal morphology during evolution.

Animals↗

Condensation of oligonucleotides assembled into nicked and gapped duplexes: potential structures for oligonucleotide delivery.

The condensation of nucleic acids into well-defined particles is an integral part of several approaches to artificial cellular delivery. Improvements in the efficiency of nucleic acid delivery in vivo are important for the development of DNA- and RNA-based therapeutics. Presently, most efforts to improve the condensation and delivery of nucleic acids have focused on the synthesis of novel condensing agents. However, short oligonucleotides are not as easy to condense into well-defined particles as gene-length DNA polymers and present particular challenges for discrete particle formation. We describe a novel strategy for improving the condensation and packaging of oligonucleotides that is based on the self-organization of half-sliding complementary oligonucleotides into long duplexes (ca. 2 kb). These non-covalent assemblies possess single-stranded nicks or single-stranded gaps at regular intervals along the duplex backbones. The condensation behavior of nicked- and gapped-DNA duplexes was investigated using several cationic condensing agents. Transmission electron microscopy and light-scattering studies reveal that these DNA duplexes condense much more readily than short duplex oligonucleotides (i.e. 21 bp), and more easily than a 3 kb plasmid DNA. The polymeric condensing agents, poly-l-lysine and polyethylenimine, form condensates with nicked- and gapped-DNA that are significantly smaller than condensates formed by the 3 kb plasmid DNA. These results demonstrate the ability for DNA structure and topology to alter nucleic acid condensation and suggest the potential for the use of this form of DNA in the design of vectors for oligonucleotide and gene delivery. The results presented here also provide new insights into the role of DNA flexibility in condensate formation.

Chlorides↗

Kinetics of nucleation controlled formation and condensational growth of disperse particles.

The kinetics of nucleation controlled formation and condensational growth of disperse particles is considered under the assumptions that: (i) only a small amount of condensable substance nucleates and forms the particles that grow by condensing the rest of the substance. (ii) The condensation efficiency is a power function of the particle mass. A nontrivial perturbation theory with respect to the smallness parameter mu= (the mass of nucleated matter)/(the total mass of condensable matter) is developed allowing one to describe the source-enhanced and free (no source) condensation processes in terms of universal functions: the particle-mass spectrum and the concentration of condensable matter. The theory relies upon a scaling transformation that removes at all the smallness parameter from the evolution equations (if the nucleation rate is a power function of the concentration of condensable matter) or leaves it in the expression for the nucleation rate where this parameter defines only a concentration scale of the nucleation process (for the nucleation rates of general form). The theory is illustrated by the exact analytical solutions of the nucleation-condensation kinetic equations for three practically important cases: (i) gas-to-particle conversion in the free-molecular regime, (ii) formation and diffusion controlled condensational growth of islands on surfaces, and (iii) formation and diffusion controlled growth of disperse particles in the continuum regime. The analytical expressions for the mass spectra of growing particles are found in the case of free condensing particles. The final mass spectra in free condensing systems display rather unusual behavior: they are either singular at small particle masses or not, depending on the value of the power exponent in the mass dependence of the condensation rate.

Journal Article↗

NTP Toxicology and Carcinogenesis Studies of Lauric Acid Diethanolamine Condensate (CAS NO. 120-40-1) in F344/N Rats and B6C3F1 Mice (Dermal Studies).

Lauric acid diethanolamine condensate is widely used in cosmetics, shampoos, soaps, and related consumer products, to which there is extensive human exposure. Because of the lack of information about potential risks associated with long-term exposure, lauric acid diethanolamine condensate, coconut oil acid diethanolamine condensate, and oleic acid diethanolamine condensate were selected as representative of the class of diethanolamides for evaluation of prechronic toxicity and carcinogenic potential. Male and female F344/N rats and B6C3F1 mice were exposed to lauric acid diethanolamine condensate dermally 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 rats were admin istered 0, 25, 50, 100, 200, or 400 mg lauric acid diethanolamine condensate/kg body weight in ethanol by dermal application for 14 weeks. All animals survived until study termination. Final mean body weights and body weight gains of males receiving 200 or 400 mg/kg were significantly less than those of the vehicle control group. Irritation of the skin at the site of application was observed in males receiving 100 mg/kg or greater and in females receiving 200 or 400 mg/kg. Kidney weights of females administered 200 or 400 mg/kg were significantly greater than those of the vehicle control group. There were dose-dependent increases in the incidences of nonneoplastic lesions of the skin at the site of application, including epidermal and sebaceous gland hyperplasia, chronic inflammation, parakeratosis, and ulcer. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were admin istered 0, 50, 100, 200, 400, or 800 mg lauric acid diethanolamine condensate/kg body weight in ethanol by dermal application for 14 weeks. All animals survived until the end of the study, and final mean body weights and body weight gains of dosed mice were generally similar to those of the vehicle control groups. Irritation of the skin at the site of application was observed in all males and females administered 400 or 800 mg/kg. The kidney weights of males receiving 100, 400, or 800 mg/kg and females receiving 800 mg/kg were significantly greater than those of the vehicle controls. Liver weights of females administered 200 mg/kg or greater were significantly greater than those of vehicle controls. Increased incidences of nonneoplastic lesions of the skin at the site of application, including epidermal and sebaceous gland hyperplasia, chronic inflammation, parakeratosis, and ulcer, were observed in males and females receiving 200 mg/kg or greater. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were admin istered 0, 50, or 100 mg lauric acid diethanolamine condensate/kg body weight in ethanol by dermal application for 104 or 105 weeks. Survival and Body Weights There were no significant differences between vehicle control and dosed males or females in survival or mean body weights. Pathology Findings There were no chemical-related differences in neoplasm incidences. Dose-related increases occurred in the incidences of nonneoplastic lesions of the skin at the site of application, including epidermal and sebaceous gland hyperplasia, hyperkeratosis, chronic inflammation, parakeratosis, and ulcer. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were admin istered 0, 100, or 200 mg lauric acid diethanolamine condensate/kg body weight in ethanol by dermal application for 105 or 106 weeks. Survival and Body Weights There were no significant differences in survival between vehicle control and dosed males or females. Mean body weights of females that received 200 mg/kg were less than those of the vehicle controls beginning at week 33. Pathology Findings The incidences of hepatocellular adenoma or carcinoma (combined) were significantly increased in dosed females compared to the vehicle controls, as was the incidence of hepatocellular adenoma in the 100 mg/kg female group. There were dose-related increases in the incidences of nonneoplastic lesions of the skin at the site of application, including epidermal and sebaceous gland hyperplasia, hyperkeratosis, chronic inflammation, and parakeratosis. Dosed males had greater incidences of thyroid gland follicular cell focal hyperplasia than did the vehicle controls. GENETIC TOXICOLOGY: Lauric acid diethanolamine condensate was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100, or TA1535, with or without S9 metabolic activation enzymes. No increase in the frequency of mutant colonies of L5178Y mouse lymphoma cells was noted after exposure to lauric acid diethanolamine condensate, with or without S9. In cytogenetic tests with cultured Chinese hamster ovary cells, lauric acid diethanolamine condensate was shown to induce sister chromatid exchanges, but not chromosomal aberrations, with and without S9. In vivo, no increase in the frequency of micro nucleated normochromatic erythrocytes was observed in peripheral blood samples from male and female mice treated dermally with lauric acid diethanolamine condensate for 14 weeks. CONCLUSIONS: Under the conditions of these 2-year dermal studies, there was no evidence of carcinogenic activity of lauric acid diethanolamine condensate in male or female F344/N rats administered 50 or 100 mg/kg or in male B6C3F1 mice administered 100 or 200 mg/kg. There was some evidence of carcinogenic activity in female B6C3F1 mice based on increased incidences of hepatocellular neoplasms. These increases were associated with free diethanolamine, which was present as a contaminant of lauric acid diethanolamine condensate. Dermal administration of lauric acid diethanolamine condensate to rats and mice for 2 years resulted in increased incidences of epidermal and sebaceous gland hyperplasia, hyperkeratosis, chronic inflammation, and parakeratosis at the site of application. Lauric acid diethanolamine condensate administration also resulted in increased incidences of thyroid gland follicular cell hyperplasia in dosed male mice. Synonyms: N,N-bis(2-hydroxyethyl) dodecanamide; N,N-bis(hydroxyethyl) lauramide; N,N-bis(b-hydroxyethyl) lauramide; bis(2-hydroxyethyl) lauramide; coco diethanolamide; coconut oil amide of diethanolamine; diethanollauramide; N,N-diethanollauramide; N,N-diethanollauric acid amide; lauramide DEA; lauric diethanolamide; lauroyl diethanolamide; lauryl diethanolamide; LDA; LDE Trade names: Clindrol 200 L; Ninol AA62; Onyxol 345; Rewomid DLMS; Rewomid DL 203/S; Richamide 6310; Rolamid CD; Standamidd LD; Steinamid DL 203 S; Super amide L-9A; Super amide L-9C; Synotol L-60; Unamide J-56; Varamid ML 1.

Journal Article↗

DNA condensation by cobalt hexaammine (III) in alcohol-water mixtures: dielectric constant and other solvent effects.

DNA molecules condense into compact structures in the presence of a critical concentration of multivalent cations. To probe the contribution of electrostatic forces to condensation, we used mixtures of water with methanol (MeOH), ethanol (EtOH), and isopropanol (iPrOH) to vary the dielectric constant epsilon from 80 to 50. The condensation of pUC18 plasmids by hexaammine cobalt (III), Co(NH3)(3+)6, was monitored by total intensity and dynamic light scattering, electron microscopy, and CD. The total scattering intensity increased as epsilon went from 80 to 70, and the decreased as epsilon decreased further. Ultraviolet spectrophotometry confirmed that the loss of intensity at low epsilon was not due to the particles' settling out of solution. The rate as well as the extent of condensation increased as epsilon was lowered from 80 to 70, and also depended on the species of alcohol (MeOH < EtOH < iPrOH). The hydrodynamic radii RH of the particles, however, remained roughly the same at 300-350 A and was independent of the species of alcohol. RH increased below epsilon = 70. The critical concentration of Co(NH3)6(3+) required to induce DNA condensation decreased from 21 microM to about 16 microM as the dielectric constant decreased from 80 to 70, and decreased moderately with the nonpolarity of the alcohol. The fraction of DNA charge neutralized at the onset of DNA condensation was calculated by a modification of Manning's two-variable counterion condensation theory to be 0.90 +/- 0.01, independent of epsilon. By electron microscopy we observed that the condensed particles changed from about 93% toroids at epsilon = 80 to 89% rods at epsilon = 70 and 98% rods at epsilon = 65. At epsilon lower than 65, DNA collapsed into a network of multistranded fibers. The morphology of condensed DNA particles, whether toroids, rods, or fibers, was independent of the alcohol species. CD spectra in ethanol-water mixtures indicated that both closed circular and linearized plasmids were in the B conformation when condensed with Co(NH3)6(3+) at epsilon > or = 70, although the closed circular molecules exhibited a weak psi-DNA spectrum. A transition from the B to A form took place between epsilon = 70 and 60, well above the normal dielectric constant of epsilon = 40 for this transition, indicating that ethanol and Co(NH3)6(3+) synergistically promote the B-A transition. We interpret these results to mean that alcohols have both electrostatic and structural effects on DNA, leading to three regimes of condensation. At the lowest alcohol concentrations the B conformation is stable and condensation is relatively slow, allowing time for the packing adjustments necessary to form toroids.(ABSTRACT TRUNCATED AT 400 WORDS)

1-Propanol↗

Molecular analysis of mitotic chromosome condensation using a quantitative time-resolved fluorescence microscopy assay.

Chromosomes condense during mitotic entry to facilitate their segregation. Condensation is typically assayed in fixed preparations, limiting analysis of contributing factors. Here, we describe a quantitative method to monitor condensation kinetics in living cells expressing GFP fused to a core histone. We demonstrate the utility of this method by using it to analyze the molecular requirements for the condensation of holocentric chromosomes during the first division of the Caenorhabditis elegans embryo. In control embryos, the fluorescence intensity distribution for nuclear GFP:histone changes during two distinct time intervals separated by a plateau phase. During the first interval, primary condensation converts diffuse chromatin into discrete linear chromosomes. After the plateau, secondary condensation compacts the curvilinear chromosomes to form shorter bar-shaped structures. We quantitatively compared the consequences on this characteristic profile of depleting the condensin complex, the mitosis-specific histone H3 kinase Aurora B, the centromeric histone CENP-A, and CENP-C, a conserved protein required for kinetochore assembly. Both condensin and CENP-A play critical but distinct roles in primary condensation. In contrast, depletion of CENP-C slows but does not prevent primary condensation. Finally, Aurora B inhibition has no effect on primary condensation, but slightly delays secondary condensation. These results provide insights into the process of condensation, help resolve apparent contradictions from prior studies, and indicate that CENP-A chromatin has an intrinsic role in the condensation of holocentric chromosomes that is independent of its requirement for kinetochore assembly.

Animals↗

Utility of exhaled breath condensates in chronic obstructive pulmonary disease: a critical review.

PURPOSE OF REVIEW: Evaluation of the utility of exhaled breath condensates in chronic obstructive pulmonary disease. RECENT FINDINGS: Exhaled breath condensates have recently been introduced as a simple, noninvasive method of sampling respiratory fluid in inflammatory lung disorders, including chronic obstructive pulmonary disease. Increases in condensate concentrations of at least 12 markers of inflammation have been reported in these disorders. Furthermore, condensate pH appears to be decreased in both chronic obstructive lung disease and bronchial asthma. This has been referred to as acidopnea and could reflect airway acidification by inflammatory cells. Although safer and more convenient than bronchoalveolar lavage, interpretation of condensate data is complicated by uncertainty regarding the source of condensate solutes and by variable dilution of respiratory droplets from condensed water vapor, which represents more than 99.9% of condensate volumes. This dilution can be estimated from the dilution of plasma constituents such as urea or electrolytes. Because the principal buffer in condensate is NH4, much of which is derived from bacterial degradation of urea in the mouth, condensate pH measurements may not provide accurate estimates of airway pH. Nevertheless, acidification of condensate may be indicative of gastroesophageal reflux, which frequently occurs in obstructive lung diseases and may contribute to cough and bronchospasm. SUMMARY: It is too early to tell how useful condensate studies will be to pulmonary investigators and clinicians. Realization of the enormous potential of this approach will require a thorough understanding of the manner in which these solutions are generated and how they should be analyzed.

Acidosis, Respiratory↗

[Breath condensate--a medium obtained by a noninvasive method for the detection of inflammation mediators of the lung].

Collection of exhaled condensate (freezing of expired air under conditions of spontaneous breathing) is a non-invasive method permitting the collection of material originating from the lung and the lower respiratory tract so that it can be used for diagnostic examinations. In order to be able to evaluate the diagnostic evidence of exhaled condensate samples in cases of respiratory disease of the calf, leukotriene B4 (LTB4) in bovine exhaled condensate was determined. The influence of the breathing pattern and body temperature on the quantity of condensate to be collected was tested in a total of 49 exhaled condensate samples. It became obvious that the exhaled condensate quantity obtained per time unit is dependent on the ventilation volume per time unit (minute volume). In exhaled condensate samples from 35 clinically healthy calves, LTB4 concentrations of less than 250 pg/mL exhaled condensate were detected. A total of 14 exhaled condensate samples from 7 calves was analyzed before and after experimental respiratory infection with Pasteurella multocida D. In parallel to the analysis of LTB4 in exhaled condensate, the lung function of the calves was examined by means of impulse oscilloresistometry. The increase of LTB4 in the exhaled condensate post infection correlated significantly (p < or = 0.05) with decreases of respiratory reactance. The determination of LTB4 concentrations in exhaled condensate seems to be suitable, in principle, for the detection of inflammations in the respiratory system of the calf. Further studies are needed for the evaluation of the diagnostic validity of the method.

Animals↗

Condensation on polymethylmethacrylate, acrylic polymer, and silicone intraocular lenses after fluid-air exchange in rabbits.

PURPOSE: Condensation occurs rapidly on silicone intraocular lenses (IOLs) after vitrectomy and fluid-air exchange in the presence of a posterior capsulotomy, severely limiting the retinal view. Foldable IOLs made of an acrylic polymer are now available. The authors compare the condensation and subsequent retinal view degradation after vitrectomy and fluid-air exchange in rabbit eyes implanted with polymethylmethacrylate (PMMA), soft acrylic polymer (ACRYSOF), and silicone IOLs. METHODS: Thirty rabbits underwent bilateral lens removal. Twenty PMMA, 20 acrylic polymer, and 20 silicone IOLs were implanted into the capsular bag. After three-port vitrectomy, eyes were randomly assigned to undergo a posterior capsulotomy or to an intact posterior capsule. Fluid-air exchange was performed, and the amount of condensation on the IOL surface and resultant retinal view was graded. Surgical maneuvers then were performed to decrease the condensation. These maneuvers included wiping the IOL surface with a soft-tipped cannula and coating it with viscoelastic. RESULTS: No condensation occurred on IOLs in eyes with an intact posterior capsule. Significantly more condensation occurred on IOLs in eyes with a capsulotomy, regardless of IOL type (P < 0.014). Wiping the condensation resulted in significantly less condensation and improved retinal view on the PMMA and acrylic IOLs when compared with the silicone IOLs (P < 0.024). Condensation was eliminated on all three IOL types after viscoelastic was applied. However, increased distortion of the retinal view was induced by the viscoelastic on the silicone IOL as compared with the other IOLs. CONCLUSIONS: Condensation and subsequent retinal image degradation occurs uniformly after vitrectomy, posterior capsulotomy, and fluid-air exchange in eyes with PMMA, acrylic, and silicone IOLs. Because the condensation can be eliminated most easily on acrylic IOLs compared with silicone IOLs, when foldable IOL techniques are used it is likely that an acrylic polymer IOL would be advantageous for those patients at risk for future vitreous procedures requiring fluid-air exchange.

Animals↗

Structure and dynamics of condensed DNA probed by 1,1'-(4,4,8,8-tetramethyl-4,8-diazaundecamethylene)bis[4-[[3- methylbenz-1,3-oxazol-2-yl]methylidine]-1,4-dihydroquinolinium] tetraiodide fluorescence.

Information on the structure and dynamics of condensed forms of DNA is important in understanding both natural situations such as DNA packaging and artificial systems such as gene delivery complexes. We have established the fluorescence of bisintercalator 1,1'-(4,4,8,8-tetramethyl-4,8-diazaundecamethylene)bis[4-[[3-methylbenz-1,3-oxazol-2-yl]methylidine]-1,4-dihydroquinolinium] tetraiodide (YOYO-1) as a novel probe for DNA condensation. When the level of DNA-bound YOYO-1 is sufficiently large, condensation by either polyethylenimine (PEI) or the cationic detergent cetyltrimethylammonium bromide (CTAB) leads to electronic interaction among YOYO-1 molecules bound on the same DNA molecule. This interaction results in an excitonic blue shift of the absorption spectra of YOYO-1 and dramatic decrease in the fluorescence quantum yield. These observations constitute a signature of the condensation of DNA. We further examined the comparative properties of DNA condensed by PEI, CTAB, or Co(NH(3))(6)(3+) through the steady-state and dynamic fluorescence of YOYO-1. Condensation by either PEI or CTAB was associated with a blue shift in the absorption spectra of YOYO-1, although the magnitude of the shift was larger in the case of PEI when compared to that of CTAB. In contrast, condensation by Co(NH(3))(6)(3+) was not associated with a measurable shift in the absorption spectra. These results were interpreted as signifying the varying level of compactness of the DNA condensates. Quenching of fluorescence by acrylamide showed that condensation by all three agents led to an increase in the level of solvent exposure of the base pairs. Observation of the decay of fluorescence intensity and anisotropy of DNA-bound YOYO-1 showed that while condensation by either PEI or CTAB froze the segmental mobility of the helix, condensation by Co(NH(3))(6)(3+) enhanced the flexibility of DNA. The relevance of our findings to functions such as efficiency of gene delivery is discussed.

Benzoxazoles↗

In-stack condensible particulate matter measurements and issues.

Particulate matter (PM) emitted from fossil fuel-fired units can be classified as either filterable or condensible PM. Condensible PM typically is not measured because federal and most state regulations do not require sources to do so. To determine the magnitude of condensible PM emissions relative to filterable PM emissions and to better understand condensible PM measurement issues, a review and analysis of actual U.S. Environmental Protection Agency (EPA) Method 202 (for in-stack condensible PM10) and EPA Method 201/201A (for in-stack filterable PM10) results were conducted. Methods 202 and 201/201A results for several coal-burning boilers showed that the condensible PM, on average, comprises approximately three-fourths (76%) of the total PM10 stack emissions. Methods 202 and 201/201A results for oil- and natural gas-fired boilers showed that the condensible PM, on average, comprises 50% of the total PM10 stack emissions. Methods 202 and 201/201A results for oil-, natural gas-, and kerosene-fired combustion turbines showed that the condensible PM, on average, comprises 69% of the total PM10 stack emissions. Based on these limited measurements, condensible PM can make a significant contribution to total PM10 emissions for fossil fuel-fired units. A positive bias (indicating more condensible PM than is actually emitted) may exist in the measured data due to the conversion of dissolved sulfur dioxide to sulfate compounds in the sampling procedure. In addition, these Method 202 results confirm that condensible PM, on average, is composed mostly of inorganic matter, regardless of the type of fuel burned.

Air Pollutants, Occupational↗

Proper chromatin condensation and maintenance of histone H3 phosphorylation during mouse oocyte meiosis requires protein phosphatase activity.

We have shown okadaic acid (OA) and calyculin-A (CLA) inhibition of mouse oocyte phosphoprotein phosphatase 1 (PPP1C) and/or phosphoprotein phosphatase 2A (PPP2CA) results in aberrant chromatin condensation, as evidenced by the inability to resolve bivalents. Phosphorylation of histone H3 at specific residues is thought to regulate chromatin condensation. Therefore, we examined changes in histone H3 phosphorylation during oocyte meiosis and the potential regulation by protein PPPs. Western blot and immunocytochemical analysis revealed histone H3 phosphorylation changed during mouse oocyte meiosis, with changes in chromatin condensation. Germinal vesicle-intact (GV-intact; 0 h) oocytes had no phospho-Ser10 but did have phospho-Ser28 histone H3. Oocytes that had undergone germinal vesicle breakdown (GVBD; 2 h) and progressed to metaphase I (MI; 7 h) and MII (16 h) had phosphorylated Ser10 and Ser28 histone H3 associated with condensed chromatin. To determine whether OA-induced aberrations in chromatin condensation were due to alterations in levels of histone H3 phosphorylation, we assessed phosphorylation of Ser10 and Ser28 residues following PPP inhibition. Oocytes treated with OA (1 microM) displayed increased phosphorylation of histone H3 at both Ser10 and Ser28 compared with controls. To begin to elucidate which OA-sensitive PPP is responsible for regulating chromatin condensation and histone H3 phosphorylation, we examined spatial and temporal localization of OA-sensitive PPPs, PPP1C, and PPP2CA. PPPC2A did not localize to condensed chromatin, whereas PPP1beta (PPP1CB) associated with condensing chromatin in GVBD, MI, and MII oocytes. Additionally, Western blot and immunocytochemistry confirmed presence of the PPP1C regulatory inhibitor subunit 2 (PPP1R2) in oocytes at condensed chromatin during meiosis and indicated a change in PPP1R2 phosphorylation. Inhibition of oocyte glycogen synthase kinase 3 (GSK3) appeared to regulate phosphorylation of PPP1R2. Furthermore, inhibition of GSK3 resulted in aberrant oocyte bivalent formation similar to that observed following PPP inhibition. These data suggest that PPP1CB is the OA/CLA-sensitive PPP that regulates oocyte chromatin condensation through regulation of histone H3 phosphorylation. Furthermore, GSK3 inhibition results in aberrant chromatin condensation and appears to regulate phosphorylation of PPP1R2.

Animals↗

Increased density of gutta-percha using a controlled heat instrument with lateral condensation.

The purpose of this study was to compare quantitatively the density of standard cold lateral gutta-percha condensation and warm lateral gutta-percha condensation using the System B heating instrument in a low-heat warm lateral condensation technique in an artificial root canal in vitro. Thirty-degree simulated root canals in 30 transparent acrylic blocks were instrumented using Gates-Glidden burs and Quantec (NT Company, Chattanooga, TN) rotary files. The canals were then obturated with gutta-percha using standard cold lateral condensation without sealer. Warm lateral condensation without sealer using the System B instrument at 101 degrees C was then performed on the same 30 canals. A second treatment of warm lateral condensation was then applied to these same canals. The blocks were weighed after the initial canal preparation and after each obturation treatment. Results showed warm lateral condensation of gutta-percha using the System B resulted in a significant increase in density by weight when compared with standard cold lateral condensation. A 23.97% increase in weight was realized after the first heat application, compared with standard lateral condensation. A second heat application produced an additional 2.59% increase in weight over that produced by the first heat application. Data were analyzed using a t test for repeated measures. Both increases were statistically significant (p < 0.001). Warm lateral condensation using the System B instrument results in denser gutta-percha fills by weight when compared with standard cold lateral condensation.

Acrylic Resins↗

Effect of condensed tannins from birdsfoot trefoil on endoglucanase activity and the digestion of cellulose filter paper by ruminal fungi.

The ruminal fungi Neocallimastix frontalis RE1, Neocallimastix patriciarum 27, Piromyces communis 22, and Orpinomyces joyonii 19-2 were examined for their ability to digest filter paper in the presence of condensed tannins from birdsfoot trefoil (Lotus corniculatus L.). For all four fungi, inhibition of endoglucanases was evident at 100 micrograms condensed tannins.mL-1 with nearly complete inhibition at 300 micrograms condensed tannins.mL-1. At 100 and 200 micrograms condensed tannins.mL-1, the endoglucanase activity of N. frontalis RE1 was greater (P < 0.01) than that of the other three fungal species. Exposure to 100 micrograms condensed tannins.mL-1 did not affect the ability of N. frontalis RE1 or N. patriciarum 27 to digest filter paper, and although digestion was reduced, N. frontalis RE1 and P. communis 22 solubilized more than 20% of the filter paper at 500 micrograms condensed tannins.mL-1. In contrast, O. joyonii 19-2 was virtually unable to digest filter paper at 300 micrograms condensed tannins.L-1. Mycelia of fungi grown with condensed tannins were covered by filamentous material, which may have arisen from the formation of condensed tannin-protein complexes. Less than 86% of the condensed tannins (as measured by the H2SO4 method) were recovered after 120 h of incubation with N. frontalis RE1, P. communis 22, and N. patriciarum 27. The need for detailed studies to examine the ability of ruminal fungi to metabolize condensed tannins is evident.

Animals↗

Condensation of DNA by trivalent cations. 2. Effects of cation structure.

Electron microscopy is employed to examine DNA aggregates produced by three tripositively charged condensing agents. Spermidine, hexammine cobalt (III), and me8spermidine (in which the amine groups of spermidine are exhaustively methylated) all produce condensates. The predominant form of condensate observed is toroidal; however, me8spermidine produces a large fraction of rodlike condensates. Distributions of toroidal radii and estimated volumes suggest that the size of condensates depends on the condensing agent employed, its concentration, and the time elapsed after addition of condensing agent. While ligand charge seems to be the major factor in predicting condensing power, ligand structure influences the morphology and dimensions of the particles produced. The ability to form hydrogen bonds is not required to promote condensation, since me8spermidine has no NHs. There may be a kinetic barrier to condensation at low me8spermidine concentrations. The relative proportions of toroids and rods may depend on the energetic compensation between bending and binding in cyclic structures, or on rate-limiting formation of sharply bent or kinked regions in rods.

Cations↗

Condensation of DNA by multivalent cations: considerations on mechanism.

DNA is generally found within viruses and cells in a tightly packaged state, typically occupying only 10(-4)-10(-6) of the volume of the uncondensed DNA wormlike coil. Condensation can be induced in vitro at low salt by the naturally occurring polyamines spermidine3+ and spermine4+, by hexammine cobalt(III), and even by Mg2+ in methanol-water mixtures. These condensates generally have an orderly, toroidal, or rodlike shape and size similar to that of DNA gently lysed from phage heads. It is also striking that the condensate size distribution is independent of DNA molecular length from 400 to 40,000 base pairs (bp), but that shorter DNA molecules (e.g., 150-bp mononucleosomal DNA) cannot condense in this fashion. We have constructed a successive association equilibrium theory to attempt to explain these results, using an equation devised by Tanford for micelle formation. Most of the obvious attractive and repulsive free energy contributions (mixing, bending, hydration, and other nearest-neighbor interactions) are linear in the amount of DNA incorporated, but the net attractive delta G0 grows nonlinearly because of the increasing average number of nearest neighbors of each duplex as the particle grows. In order that the size distribution have a maximum, a quadratic repulsive free energy is also required, arising from the electrostatic self-energy of the incompletely neutralized particles. The net attractive free energy per base pair interaction is tiny, on the order of 10(-3) kT. Despite the apparent generally correct order of magnitude of the various free energy terms, the calculated size distribution is smaller and narrower than observed experimentally. It appears that the size distribution of condensed particles is determined kinetically rather than thermodynamically. Very short DNA molecules cannot nucleate stable aggregates because they cannot develop adequate overlap, either internally or intermolecularly. A substantial fraction of rodlike condensates is observed in aqueous solutions only with a rather inefficient condensing agent, permethylated spermidine. This suggests that slow condensation kinetics may be required to overcome the high activation energy of highly distorted DNA bends or kinks at the turning points of rods. Evidence is reviewed that condensation may be associated with localized helix structure distortion provoked by condensing agents.

DNA↗