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Induction of structural and numerical changes of chromosome, centrosome abnormality, multipolar spindles and multipolar division in cultured Chinese hamster V79 cells by exposure to a trivalent dimethylarsenic compound.

Dimethylarsine iodide (DMI) was used as a model compound of trivalent dimethylarsenicals [DMA(III)], and the biological effects were extensively investigated in cultured Chinese hamster V79 cells. When the cytotoxic effects of DMA(III) were compared with those of inorganic arsenite and dimethylarsinic acid [DMA(V)], DMA(III) was about 10,000 times more potent than DMA(V), and it was even 10 times more toxic than arsenite. Depletion of cell glutathione (GSH) did not influence the cytotoxic effects of DMA(III), whereas it enhanced the cytotoxicity of arsenite. Chromosome structural aberrations, such as gaps, breaks and pulverizations, and numerical changes, such as aneuploidy, hyper- and hypo-tetraploidy, were induced by DMA(III) in a concentration-dependent manner. Mitotic index increased 9-12h after the addition of DMA(III), and then declined. By contrast, the incidence of multinucleated cells increased conversely with the decrease in mitotic index at and after 24h of exposure. The mitotic cell-specific abnormality of centrosome integrity and multipolar spindles were induced by DMA(III) in a time- and concentration-dependent manner. Moreover, DMA(III) caused abnormal cytokinesis (multipolar division) at concentrations that were effective in causing centrosome abnormality, multipolar spindles and aneuploidy. These results showed that DMA(III) was genotoxic on cultured mammalian cells. Results also suggest that DMA(III)-induced multipolar spindles and multipolar division may be associated with the induction of aneuploidy. In addition, the centrosome may be a primary target for cell death via multinucleated cells.

Aneuploidy↗

Protection against radiation-induced chromosome damage in mouse bone marrow by Ocimum sanctum.

The radioprotective effect of the leaf extract of Ocimum sanctum (Ocimum extract, OE) was investigated by taking chromosome aberrations as the end point. Adult Swiss mice were whole-body exposed to 1-6 Gy of gamma radiation with/without pretreatment with 10 mg/kg b.wt. of OE intraperitoneally for 5 consecutive days. Radiation was given 30 min after the last injection. Metaphase plates were prepared from femur marrow on days 1, 2, 7 and 14 post-treatment and the frequency of aberrant cells and individual aberrations were scored. OE alone did not have any significant effect on the chromosomes. Maximum percent of aberrant cells was observed at 24 h in all the exposed groups. The percent aberrant cells showed a linear quadratic increase with radiation dose, in both radiation alone (RT) and OE + RT-treated groups. Exchange (dicentrics and rings) and multiple (pulverized and severely damaged cells) aberrations also showed a similar response. However, the slopes of OE + RT was significantly shallower than RT groups (p < 0.05). A dose-modifying factor of 2.63 was obtained taking percent aberrant cells for 2 Gy as the base. Progressive decline in the percent aberrant cells as well as the number of aberrations with time after irradiation was observed in both RT and OE + RT groups. OE treatment resulted in a faster recovery compared to RT alone group. At doses below 3 Gy, OE pretreatment almost completely eliminated the exchange aberrations from the cell population by day 2. Studies on a chemical system demonstrated that OE significantly reduced the generation of hydroxyl radical; a lower dose of OE (1 mg/ml) was more effective than 5 mg/ml and this effect was more pronounced than that produced by DMSO. These results show that OE affords in vivo protection against radiation-induced cytogenetic damage. Free radical scavenging is a likely mechanism of OE protection.

Animals↗

Enhancement of bone marrow radioprotection and reduction of WR-2721 toxicity by Ocimum sanctum.

The radioprotective effect of the leaf extract of Ocimum sanctum (OE) in combination with WR-2721 (WR) was investigated on mouse bone marrow. Adult Swiss mice were injected intraperitoneally (i.p.) with OE (10 mg/kg on 5 consecutive days), or 100-400 mg/kg WR (single dose) or combination of the two or double-distilled water (DDW) and whole-body exposed to 4.5 Gy gamma-irradiation (RT). Metaphase plates were prepared from femur bone marrow on days 1, 2, 7 and 14 post-treatment and chromosomal aberrations were scored. The maximum number of aberrant cells was observed at 24 h after irradiation in all the groups. However, pretreatment with OE or WR individually resulted in a significant decrease in aberrant cells as well as different types of aberrations. The combination of the two further enhanced this effect; resulting in a 2-fold increase in the protection factor (PF = 6.68) compared to 400 mg/kg WR alone. The percent aberrant cells decreased linear-quadratically with WR dose when given individually, while in the OE + WR pretreatment animals the values showed a linear dose response. Combination of OE with WR doses above 200 mg/kg completely eliminated rings, polyploidy and pulverization of chromosomes. Percent aberrant cells decreased with time in all groups, though the values remained higher than normal even on day 14 in the RT alone as well as those treated with single agent + RT. WR doses above 200 mg/kg before RT resulted in significantly higher frequency of aberrant cells compared to RT and OE + RT groups on day 14, suggesting delayed WR toxicity; but combination of OE with WR brought down these values to normal level, indicating that OE combination, in addition to enhancing WR protection, may also act as a detoxifier. The protective effect of OE and WR is also reflected in the enhancement of bone marrow CFU survival. Both OE and WR possessed significant free radical scavenging activity in vitro. The combination of the two further enhanced this effect, suggesting that the enhanced free radical scavenging activity by combining the two protectors results in the higher bone marrow cell protection. The significant elevation in chromosome protection obtained by combining OE with WR, with reduction in the latter's toxicity at higher doses, suggests that the combination may have promise for radioprotection in humans.

Amifostine↗

PAH and soot emissions from burning components of medical waste: examination/surgical gloves and cotton pads.

This is a laboratory investigation on the emissions from batch combustion of representative infectious ("red bag") medical waste components, such as medical examination latex gloves and sterile cotton pads. Plastics and cloth account for the majority of the red bag wastes by mass and, certainly, by volume. An electrically heated, horizontal muffle furnace was used for batch combustion of small quantities of shredded fuels (0.5-1.5 g) at a gas temperature of approximately 1000 degrees C. The residence time of the post-combustion gases in the furnace was approximately 1 s. At the exit of the furnace, the following emissions were measured: CO, CO2, NOx, particulates and polynuclear aromatic compounds (PACs). The first three gaseous emissions were measured with continuous gas analyzers. Soot and PAC emissions were simultaneously measured by passing the furnace effluent through a filter (to collect condensed-phase PACs) and a bed of XAD-4 adsorbent (to capture gaseous-phase PACs). Analysis involved soxhlet extraction, followed by gas chromatography-mass spectrometry (GC-MS). Results were contrasted with previously measured emissions from batch combustion of pulverized coal and tire-derived fuel (TDF) under similar conditions. Results showed that the particulate soot) and cumulative PAC emissions from batch combustion of latex gloves were more than an order of magnitude higher than those from cotton pads. The following values are indicative of the relative trends (but not necessarily absolute values) in emission yields: 26% of the mass of the latex was converted to soot, 11% of which was condensed PAC. Only 2% of the mass of cotton pads was converted to soot, and only 3% of the weight of that soot was condensed PAC. The PAC yields from latex were comparable to those from TDF. The PAC yields from cotton were higher than those from coal. A notable exception to this trend was that the three-ring gas-phase PAC yields from cotton were more significant than those from latex. Emission yields of CO and CO2 from batch combustion of cotton were, respectively, comparable and higher than those from latex, despite the fact that the carbon content of cotton was half that of latex. This is indicative of the more effective combustion of cotton. Nearly all of the mass of carbon of cotton gasified to CO and CO2 while only small fractions of the carbon in latex were converted to CO2 and CO (20% and 10%, respectively). Yields of NOx from batch combustions of latex and cotton accounted for 15% and 12%, respectively, of the mass of fuel nitrogen indicating that more fuel nitrogen was converted to NOx in the former case, possibly due to higher flame temperatures. No SO2 emissions were detected, indicating that during the fuel-rich combustion of latex, its sulfur content was converted to other compounds (such as H2S) or remained in the soot.

Carbon↗

Inhalation health effects of fine particles from the co-combustion of coal and refuse derived fuel.

This paper is concerned with health effects from the inhalation of particulate matter (PM) emitted from the combustion of coal, and from the co-combustion of refuse derived fuel (RDF) and pulverized coal mixtures, under both normal and low NO(x) conditions. Specific issues focus on whether the addition of RDF to coal has an effect on PM toxicity, and whether the application of staged combustion (for low NO(x)) may also be a factor in this regard. Ash particles were sampled and collected from a pilot scale combustion unit and then re-suspended and diluted to concentrations of approximately 1000 microg/m(3). These particles were inhaled by mice, which were held in a nose-only exposure configuration. Exposure tests were for 1 h per day, and involved three sets (eight mice per set) of mice. These three sets were exposed over 8, 16, and 24 consecutive days, respectively. Pathological lung damage was measured in terms of increases in lung permeability. Results show that the re-suspended coal/RDF ash appeared to cause very different effects on lung permeability than did coal ash alone. In addition, it was also shown that a "snapshot" of lung properties after a fixed number of daily 1-h exposures, can be misleading, since apparent repair mechanisms cause lung properties to change over a period of time. For the coal/RDF, the greatest lung damage (in terms of lung permeability increase) occurred at the short exposure period of 8 days, and thereafter appeared to be gradually repaired. Ash from staged (low NO(x)) combustion of coal/RDF appeared to cause greater lung injury than that from unstaged (high NO(x)) coal/RDF combustion, although the temporal behavior and (apparent) repair processes in each case were similar. In contrast to this, coal ash alone showed a slight decrease of lung permeability after 1 and 3 days, and this disappeared after 12 days. These observations are interpreted in the light of mechanisms proposed in the literature. The results all suggest that the composition of particles actually inhaled is important in determining lung injury. Particle size segregated leachability measurements showed that water soluble sulfur, zinc, and vanadium, but not iron, were present in the coal/RDF ash particles, which caused lung permeabilities to increase. However, the differences in health effects between unstaged and staged coal/RDF combustion could not be attributed to variations in pH values of the leachate.

Air Pollutants↗

Resuspension of coal and coal/municipal sewage sludge combustion generated fine particles for inhalation health effects studies.

Airborne particulate matter (PM) is an important environmental issue because of its association with acute respiratory distress in humans, although the specific particle characteristics that cause lung damage have yet to be identified. Particle size, acid aerosols, water-soluble transition metals (e.g. Cu, Fe, V, Ni and Zn), polyaromatic hydrocarbons, and particle composition are the focus of several popular hypotheses addressing respiratory distress. All of the above mentioned characteristics are contained in PM generated from the combustion of both pulverized coal, and biomass, including dried municipal sewage sludge (MSS). In this investigation, we report results from collaborative interdisciplinary research on the inhalation health risks caused by particles emitted from the co-combustion of municipal sewage sludge (MSS) and coal. A solid particle resuspension system was implemented to resuspend ash particles. Mice were exposed to resuspended coal and MSS/coal ash particles. Mice exposed to MSS/coal ash particulate demonstrated significant increases in lung permeability, a marker of the early stages of pathological lung injury, while the mice exposed to coal-only ash did not. These results show that the composition of particles actually inhaled is important in determining lung damage. Zinc was significantly more concentrated in the MSS/coal ash than coal ash particles and the pH of these particles did not differ significantly. Specifically, an MSS/coal mixture, when burned, emits particles that may cause significantly more lung damage than coal alone, and that consequently, the use of MSS as a 'green', CO2-neutral replacement fuel should be carefully considered.

Air Pollutants↗

Development of a new calcium phosphate cement that contains sodium calcium phosphate.

A cement powder consisting of sodium calcium phosphate, Na3Ca6(PO4)5, in addition to tetracalcium phosphate and beta-tricalcium phosphate was prepared by pulverizing blocks of 4 wt% sodium-, 11 wt% carbonate-containing apatite samples that were heated at 1700 degrees C for 5 h. When mixed with 30 wt% malic acid or citric acid at a powder liquid ratio of 3:1, the cement set in 3 or 7 min at room temperature with compressive strength being around 52 or 27 MPa. In HeLa-cell cultures, the cement mixed with malic acid was less cytotoxic than the cement mixed with citric acid, which was far less cytotoxic than a commercial carboxylate cement used as a negative control, suggesting malic acid to be superior to citric acid as a liquid in this regard. Similar findings were also obtained with osteoclasts, of which culture experiments clearly suggested that the number of osteoclasts on the cement mixed with malic acid was significantly greater than that on the cement mixed with citric acid. Since osteoclastic response to substrates could be used as a maker in evaluating their bioresorbability associated with osteoclasts, the above finding may suggest that the cement that is to be mixed with malic acid would be more useful as bone substitutes.

Animals↗

Preparation and histological evaluation of biomimetic three-dimensional hydroxyapatite/chitosan-gelatin network composite scaffolds.

A novel biodegradable hydroxyapatite/chitosan-gelatin network (HA/CS-Gel) composite of similar composition to that of normal human bone was prepared as a three-dimensional biomimetic scaffold by phase separation method for bone tissue engineering. Changing the solid content and the compositional variables of the original mixtures allowed control of the porosities and densities of the scaffolds. The HA granules were dispersed uniformly in the organic network with intimate interface contact via pulverizing and ultrasonically treating commercial available HA particles. Scaffolds of 90.6% porosity were used to examine the proliferation and functions of the cells in this three-dimensional microenvironment by culturing neonatal rat caldaria osteoblasts. Histological and immunohistochemical staining and scanning electron microscopy observation indicated that the osteoblasts attached to and proliferated on the scaffolds. Extracellular matrices including collagen I and proteoglycan-like substrate were synthesized, while osteoid and bone-like tissue formed during the culture period. Furthermore, the cell/scaffold constructs had good biomineralization effect after 3 weeks in culture.

Animals↗

Antibacterial silver-containing silica glass prepared by sol-gel method.

Recently, various inorganic antibacterial materials containing silver have been developed and some of them are in commercial use. Colorless and more chemically durable materials which slowly release the silver ion for a long period are, however, desirable to be developed for medical applications such as composite resin for dental restoration. In the present study, Si(OC2H5)4, Al(NO3)3 x 9H2O, AgNO3, HNO3, C2H5OH and H2O solutions with various Al/Ag atomic ratios under a constant Si/Ag atomic ratio of 1/0.023 were kept at 40 degrees C for gelation and drying. Thus obtained gels were pulverized into fine powders with average particle size of approximately 10 microm and then heat-treated at 900-1000 degrees C for 2 h. For the composition Al/Ag = 0, a yellow-colored glass was formed, since the silver existed in the form of metallic colloids in the glass. However, for the compositions Al/Ag > or = 1, colorless glasses were successfully obtained, since the silver existed in the form of Ag+ ions in the glasses. For the composition Al/Ag = 0, the silver ions got released rapidly into the water, whereas, for the compositions Al/Ag > or = 1, they gradually got released into the water at a controlled rate. A composite of the obtained powders with Al/Ag atomic ratio of 1 with Bis-GMA/TEGDMA in 70:30 weight ratio showed excellent antibacterial property. The sol-gel derived silica glass powders containing silver with compositions Al/Ag > or = 1 are believed to be useful as an antibacterial material for medical applications such as filler of composite resin for dental restoration.

Aluminum Compounds↗

The effect of neodymium:YAG laser shocks on the blood-aqueous barrier.

An animal model demonstrates that the acute inflammation seen after neodymium: YAG (Nd:YAG) capsulotomy is related to the presence of disrupted tissue suspended in the aqueous, rather than to the mechanical insults by the repeated shock waves. Seven rabbits were treated in the lens cortex of one eye with 20 bursts of 4 pulses, 24 mJ each, and followed fluorophotometrically using albumin labelled with fluorescein. This allowed transmission of shock waves to the anterior segment without releasing debris in five eyes that showed no inflammation. Two eyes with inadvertent capsular rupture showed marked blood-aqueous barrier breakdown. This suggests the use of a capsulotomy technique that relies on discission rather than pulverization of the membrane and avoids the thicker portions of the membrane when possible.

Animals↗

No chromosomal instability in offspring of survivors of childhood malignancy.

Chromosomal instability was examined in 20 apparently healthy children of survivors of childhood malignancy. As compared to controls, no increase of spontaneous or bleomycin-induced aberrations (including gaps, breaks, sister chromatid exchanges, pulverization, and premature centromere divisions) were found in these "index children." The results suggest that the offspring of subjects previously receiving chemotherapy and/or radiotherapy for childhood malignancy are probably at no increased risk of latent chromosomal instability.

Bleomycin↗

A small-scale procedure for extracting nucleic acids from woody plants infected with various phytopathogens for PCR assay.

The complexity of most nucleic acid extraction procedures limits the number of samples that can be easily processed for analysis by polymerase chain reaction (PCR). A simple, small-scale procedure was developed which can be carried out entirely in 1.5-ml microfuge tubes whereby the container and contents are frozen with liquid nitrogen, tissue is pulverized, and targeted nucleic acids are extracted. DNA of bacterial and phytoplasmal plant pathogens was extracted in hot CTAB buffer followed by chloroform clarification. Following centrifugation, the DNA in the aqueous fraction was precipitated with isopropanol and resuspended in water. RNA originating from viruses and viroids was extracted from triturated tissue using STE buffer and phenol. The nucleic acid fraction was purified using CF-11 cellulose. All purified preparations were used as PCR or RT-PCR templates to detect DNA or RNA, respectively. These procedures were used to detect Xylella fastidiosa, peach yellow leaf roll phytoplasma, sour cherry green ring mottle virus, and peach latent mosaic viroid by agarose gel electrophoresis.

DNA, Bacterial↗

Dietary xylitol prevents ovariectomy induced changes of bone inorganic fraction in rats.

Thirty-six 3-month-old female Wistar rats were labelled with a single intraperitoneal tetracycline injection. Twenty-four animals were subsequently ovariectomized, while the control group of 12 animals underwent sham operations. All animals received the basal Ewos R3 diet and half of the ovariectomized animals (n = 12) were given an additional 5% dietary xylitol supplementation. Three months later, following the collection of blood and urine, the animals were killed by decapitation. The tibiae were detached and prepared for chemical and other studies. The weight and density of the tibiae were measured. The right tibiae were dried and pulverized for chemical analysis of calcium, phosphorus and citric acid. The total inorganic fraction was determined by ashing the powdered bone. The left tibiae were cross-sectioned at the tibio-fibular junctions for the measurement of the width of periosteally formed bone, which was identified by tetracycline fluorescence. The examination of mineral content of bone was performed by scanning electron microscopy, using an electron probe microanalytic technique. The results indicate that the supplementation of the diet with 5% xylitol had a protective effect against the loss of bone mineral after ovariectomy in the rat. This was clearly seen in tibial density and in the inorganic fraction of the bone, and in the concentrations of bone Ca and phosphorus. Ovariectomy caused a doubling in periosteal bone formation relative to the controls, whereas the growth of the periosteally formed bone was somewhat reduced following xylitol supplementation as compared with ovariectomy alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Observation and characterization of colloids derived from leached cement hydrates.

The possibility of colloid generation from cement hydrates in a cementitious repository environment has been investigated through leaching experiments. Pulverized samples of High Flyash and Silica fume-content Cement (HFSC) and 1:9 ordinary portland cement/blast furnace slag (1:9 OPC/BFS) hydrate were leached in low-salinity groundwater at three solid-to-liquid (S/L) mass ratios (1:5, 1:50 and 1:100), and two temperatures (20 and 60 degrees C) for durations of nearly 2 and 8 months. Detailed characterization of colloid populations has been undertaken by TEM coupled with X-ray analysis. In addition, the surface charge and stability behavior of colloids have been investigated. The colloid concentrations in HFSC hydrate leachates generated at 20 and 60 degrees C show similar trends with S/L ratio. The colloid concentrations of leachates with the lower S/L ratio (1:50 and 1:100) are in the range of 10(11)-10(12) particles per liter. The majority of these particles are composed predominantly of Si, Ca, and Al; the mean particle size is less than 100 nm. The lowest colloid concentrations are found in the leachates with the highest S/L ratios, and the colloid populations tend to be dominated by larger particles. HFSC-derived colloid stability is due to a high negative zeta potential at alkaline pH values, combined with a calcium concentration that is below the critical coagulation concentration (CCC) for the colloids. A preliminary interpretation of HFSC-derived colloid stability based on classical DLVO theory provides a semi-quantitative explanation of the dependence of colloid populations on the S/L ratio in the leaching experiments.

Colloids↗

Factitial cutaneous ulcers and nodules: the use of electron-probe microanalysis in diagnosis.

Two cases are reported in which persistent nodular or ulcerative skin lesions with granulomatous histology suggested possible injection of foreign material. Electron-probe microanalysis was used to determine the presence and nature of foreign materials present in the skin. This technic identified specific agents used as fillers in tablets, indicating the injection of pulverized tablet material. The patients were counseled regarding this evidence, and psychotherapy and drug rehabilitation were recommended in each instance. This technic provides a valuable tool for accurate identification of foreign substances in the skin.

Adult↗

Adhesion, spreading and growth of animal cells on the surface of glass ceramic Ap40--a contribution to the cell compatibility of dental permanent hard tissue implants.

Cells of different cell lines on the surface of glass ceramic Ap40 responded in the manner of reversible vacuolization in the cytoplasm. Whereas the eluate of bulk glass ceramic did not produce toxic effect on cell populations in vitro, the eluate of pulverized glass ceramic Ap40 caused a toxic reaction, the effect being dependent on concentration and incubation time. Although cell attachment, spreading and proliferation of fibroblast-like and epithel-like cells on glass ceramic surfaces are delayed when compared to glass and polystyrene the cells are adhering to a confluent monolayer.

Animals↗

Pig chromosome aberrations after vaccination against classical swine fever in field trials.

Chromosomal aberrations were observed after vaccination against classical swine fever (CSF) in a previous study done on experimental pigs. To determine if the same effect occurs in farm animals, field trials were done with immunized pigs. The cytogenetic analysis was made from lymphocyte cultures of pigs sampled in three farms of Río Cuarto region on six different periods: one, pre-vaccination (control) and in five post-vaccination (days 3, 7, 10, 15 and 22). Vaccine inoculation induced significant increase of cell frequency with chromosomal aberrations from days 3 to 10, when maximal frequencies of 6.78, 10.36 and 7.21% were observed in farms A, B and C, respectively. Afterwards, a decrease was recorded, reaching values not significantly different from controls. Mean frequencies of cells with chromosomal aberrations were 2.8+/-0.3, 5.9+/-0.4, 3.1+/-0.3%, for A, B and C farms, respectively. Cytogenetic effect was clastogenic, being chromosome breaks the first type of alteration to increase from the day 3 post-vaccination onwards, followed by chromatid exchanges and cells with multiple abnormalities. Chromosome pulverization was the last type to show increment, reaching a top value on day 10, after that it started to diminish gradually. There was no difference in the frequency of polyploid cells among sampling dates meaning that this type of alteration would not be induced by vaccination. Results of this field study confirm the mutagenic capacity of live virus vaccines against CSF and indicate that the evolution of chromosome alterations on the five evaluated post-vaccination periods was similar at different environments.

Animals↗

The experimental study of microbial contamination of the space hardware.

The role of potential contaminants of design materials and products of space technology--aerobic and anaerobic prokaryotes (myxobacteria, eubacteria, corinebacteria, actinomyces), and eukaryotes (micromyces), psychrophilic, mesophilic and thermophilic forms, chemolythotrophic microorganisms is discussed in this paper. The methods of analysis of microbial contamination in the solution of problem of the planetary protection are considered. The necessity of the use of ultrasound at the evaluation of surface and subsurface contamination of specimens is demonstrated; methods of determination of buried contamination (with the use of organic solvents and mechanical pulverization) are discussed. The data on buried and subsurface contamination for some materials and electronic parts together with microflora resistivity to sterilizing treatment are given.

Bacteria↗