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Retrospective dose reconstruction of human radiation exposure by FISH/chromosome painting.

Measurements of stable chromosomal aberrations by fluorescence in situ hybridisation (FISH)-chromosome painting has the potential to be used for dose reconstruction, years after exposure to ionising radiation. The method is, however, not yet fully standardised and validated. In particular, with respect to the limited lifespan of circulating T-lymphocytes, a level of uncertainty exists on the long-term persistence of stable aberrations. The main principles of the technique will be demonstrated. Based on results from the literature, the reliability and limitations are discussed.

Calibration↗

Characterization of chromosome changes in two human prostatic carcinoma cell lines (PC-3 and DU145) using chromosome painting and comparative genomic hybridization.

Using chromosome painting, a study of chromosomal abnormalities has been performed in two prostatic carcinoma cell lines, PC-3 and DU145. In PC-3, this analysis revealed a highly rearranged hypotriploid karyotype with 54 to 61 chromosomes and numerous rearrangements of chromosomes 1, 3, 5, 8, 10, and 14. At passage 73, DU145 had a hypotriploid karyotype with few rearrangements of chromosomes 1, 3, 5, 12, 13, and 20, whereas at passage 153, this cell line showed a near-tetraploid karyotype with a great number of rearrangements involving chromosomes 3, 6, 8, 10, 12, and 17. A single rearrangement was shared by the 2 cell lines, an i(5)(p10). A comparative genomic hybridization study demonstrated a noticeable amplification of bands 10q22.3-q23 and 14q22-q24 in the PC-3 cell line. No amplification signal was detected for DU145.

Carcinoma↗

Toxicologic evaluation of flavor ingredients added to cigarette tobacco: skin painting bioassay of cigarette smoke condensate in SENCAR mice.

Four comparative two-stage SENCAR mouse skin painting bioassays were conducted with cigarette smoke condensate (CSC) preparations to evaluate the effect of common American cigarette flavoring ingredients on tumor promotion. Each independent study employed a unique flavoring combination applied to tobacco at exaggerated levels, and in total resulted in an evaluation of 150 ingredients. Groups of 30-50 female SENCAR mice each were initiated topically with 50 microg of 7,12-dimethylbenz(a)anthracene (DMBA), and promoted thrice weekly for 26 weeks with either 10 or 20 mg of CSC from test cigarettes containing ingredient mixtures. For comparison, separate groups of mice received concurrent treatment with CSC from reference cigarettes prepared without added ingredients. Negative and positive controls were treated with acetone or 12-0-tetradecanoyl-phorbol-13-acetate (TPA) as a promoter, respectively. CSC-only groups served as promotion controls. Tumors developed in > 80% of the TPA-treated mice by study week 11, with a < 3% background tumor formation in the acetone treated controls at termination. Tumor incidence in CSC-only promotion control groups was < 20%, with no apparent difference between reference and test CSC groups. Approximately 70% of the DMBA-initiated mice promoted with 20 mg CSC developed tumors. Tumors first appeared around week 9, with about five tumors/tumor bearing animal. Tumor incidence, latency and multiplicity were CSC dose related, with a lower tumor incidence (approximately 50%), longer latency (12 weeks), and reduced tumor burden (four tumors/tumor bearing animal) at the 10 mg CSC dose level. While tumor incidence, latency and multiplicity data occasionally differed between test and comparative reference CSC groups, all effects appeared to be within normal variation for the model system. Furthermore, none of the changes appeared to be substantial enough to conclude that the tumor promotion capacity of CSC obtained from cigarettes containing tobacco with ingredients was discernibly different from the CSC obtained from reference cigarettes containing tobacco processed without ingredients.

9,10-Dimethyl-1,2-benzanthracene↗

Characteristics of nest soil, but not geographic origin, influence cold hardiness of hatchling painted turtles.

We investigated environmental factors influencing cold hardiness in hatchling painted turtles (Chrysemys picta) indigenous to northeastern Indiana and the Sandhills of west-central Nebraska. In both locations, hatchlings overwinter in their natal nests. Survival of hatchlings chilled to minimum temperatures between -2.5 and -6.0 degrees C inside explanted natal nests ranged from 30 to 100%. Mortality likely was caused by freezing of the turtles that was induced by contact with ice nuclei in the surrounding soil. Susceptibility to inoculative freezing was strongly influenced by moisture content (7.5-25%, w/w) of the frozen soil in which hatchlings were cooled. When chilled in soil containing 15% moisture, turtles from Indiana resisted inoculative freezing better than hatchlings from Nebraska, but this variation was due to physical characteristics of the soils indigenous to each locale rather than genetic differences between populations. Soil in which the Indiana turtles nested contained relatively higher amounts of clay and organic matter, and bound more moisture, than the loamy sand at the Nebraska site. Soil collected from both locales contained potent ice nuclei that may constrain supercooling of the hatchlings, even in the absence of soil moisture. In addition to temperature and precipitation, local and regional variation in soils is an important determinant of overwintering survival of hatchling C. picta.

Journal Article↗

Natural freeze-tolerance in hatchling painted turtles?

Hatchlings of the North American painted turtle (Family Emydidae: Chrysemys picta) typically spend their first winter of life inside a shallow, subterranean hibernaculum (the natal nest) where life-threatening conditions of ice and cold commonly occur. Although a popular opinion holds that neonates exploit a tolerance for freezing to survive the rigors of winter, hatchlings are more likely to withstand exposure to ice and cold by avoiding freezing altogether-and to do so without the benefit of an antifreeze. In the interval between hatching by turtles in late summer and the onset of wintery weather in November or December, the integument of the animals becomes highly resistant to the penetration of ice into body compartments from surrounding soil, and the turtles also purge their bodies of catalysts for the formation of ice. These two adjustments, taken together, enable the animals to supercool to temperatures below those that they routinely experience in nature. However, cardiac function in hatchlings is diminished at subzero temperatures, thereby compromising the delivery of oxygen to peripheral tissues and eliciting an increase in reliance by those tissues on anaerobic metabolism for the provision of ATP. The resulting increase in production of lactic acid may disrupt acid/base balance and lead to death even in animals that remain unfrozen. Although an ability to undergo supercooling may be key to survival by overwintering turtles in northerly populations, a similar capacity to resist inoculation and undergo supercooling characterizes animals from a population near the southern limit of distribution, where winters are relatively benign. Thus, the suite of characters enabling hatchlings to withstand exposure to ice and cold may have been acquired prior to the northward dispersal of the species at the end of the Pleistocene, and the characters may not have originated as adaptations specifically to the challenges of winter.

Acclimatization↗

Hatchling painted turtles (Chrysemys picta) survive only brief freezing of their bodily fluids.

Neonatal painted turtles (Chrysemys picta) spend their first winter inside the shallow, subterranean nest cavity where they completed embryogenesis. Consequently, hatchlings at high latitudes may be exposed to ice and cold during the winter. This study was undertaken to determine how long hatchlings withstand freezing at temperatures slightly below 0 degree C because tolerance for freezing has been proposed to be the key to survival by overwintering animals. A thermocouple was glued to the carapace of each hatchling. The animal was dipped in water to provide a site of nucleation of ice and was then placed into a glass jar that was partially immersed in a circulating bath at -2 degrees C. Carapace temperature was monitored throughout the procedure. When a freezing exotherm was detected, timing of the freezing event began. Animals were maintained in a frozen state for 12-48 h prior to being warmed to room temperature. Of the 39 hatchlings, 22 did not survive, and mortality increased as the duration of freezing increased. Logistic regression indicates that no turtle would have survived in a frozen state for more than 54 h. These results indicate that hatchlings can survive only brief exposure to freezing of the body fluids. Thus, hatchlings cannot tolerate freezing during prolonged periods of cold.

Acclimatization↗

Theragnostic imaging for radiation oncology: dose-painting by numbers.

Theragnostic imaging for radiation oncology is the use of molecular and functional imaging to prescribe the distribution of radiation in four dimensions-the three dimensions of space plus time-of radiotherapy alone or combined with other treatment modalities in an individual patient. Several new imaging targets for positron-emission tomography, single-photon-emission CT, and magnetic resonance spectroscopy allow variations in microenvironmental or cellular phenotypes that modulate the effect of radiation to be mapped in three dimensions. Dose-painting by numbers is a strategy by which the dose distribution delivered by inverse planned intensity-modulated radiotherapy is prescribed in four dimensions. This approach will revolutionise the way that radiotherapy is prescribed and planned and, at least in theory, will improve the therapeutic outcome in terms of local tumour control and side-effects to unaffected tissue.

Diagnostic Imaging↗

Cadmium: tissue distribution and binding protein induction in the painted turtle, Chrysemys picta.

The freshwater painted turtle, Chrysemys picta, was used to investigate (a) the distribution of an injected dose of 109Cd in tissues over a period of 192 h (8 days) and (b) the effect of non-isotopic cadmium injection on tissue metal-binding protein levels. Cadmium is cleared from the blood with 9% remaining in the circulation at 192 h. 109Cd is found in all tissues, but is accumulated preferentially in liver, kidney, pancreas, and gastrointestinal tract. The liver is the primary site of Cd accumulation, accounting for 46.4% of the injected dose by 192 h and the highest Cd concentration (cpm/mg tissue). Steroidogenic tissues and the oviduct accumulate significant amounts of 109Cd and the isotope is present in yolk. An increase in tissue metal-binding protein level after non-isotopic CdCl2 injection is consistent with 109Cd distribution, in that metal-binding protein concentration after CdCl2 injection is highest in liver, followed by pancreas and kidney with low, but with significant levels of cadmium-binding protein in gonads and steroid target organs. We conclude that the liver is the major site of storage after a single injection of isotopic cadmium and induction of a metal-binding protein may be an adaptive response to exposure to cadmium.

Animals↗

One-step, painting-like coating procedures to make surfaces highly and permanently bactericidal.

Previously we found that covalent attachment of long-chained, moderately hydrophobic polycations to surfaces of solid objects renders the latter permanently bactericidal. Herein we replaced such surface-specific, multistep immobilization techniques with a single-step, general procedure akin to common painting. Glass or polyethylene slides were briefly dipped into organic solutions of certain optimally hydrophobic N-alkyl-PEI (where PEI stands for branched 750-kDa polyethylenimine) polycations, followed by solvent evaporation. The resultant polycation-coated slides were able to kill on contact all of the encountered bacterial cells, whether the Gram-positive human pathogen Staphylococcus aureus or its Gram-negative brethren Escherichia coli. This biocidal effect was found not to be caused by N-alkyl-PEI molecules leached from the surface. Further examination of the mechanism of this bactericidal action suggested that the surface-deposited N-alkyl-PEI kills bacteria by rupturing their cellular membranes. This conclusion was further supported by studies in which the molecular weight of PEI and the hydrophobicity of the alkyl moiety were varied.

Escherichia coli↗

Fast autoxidation of ethyl linoleate catalyzed by [Mn(acac)3] and bipyridine: a possible drying catalyst for alkyd paints.

It has been found that [Mn(acac)(3)], by functioning both as a radical initiator and a hydroperoxide decomposition catalyst, has a very high activity in the autoxidation of ethyl linoleate (EL), a model compound for the binder molecule in household alkyd paint. Adding 1 equiv of bipyridine (bpy) to a reaction mixture of EL and [Mn(acac)(3)] significantly enhances the autoxidation rate. The redox properties of [Mn(acac)(3)] and [Mn(acac)(2)(bpy)] have been compared with cyclic voltammetry; the observed peak potentials show that addition of bpy facilitates reduction of Mn(III) to Mn(II), which may explain the increase in autoxidation activity.

Journal Article↗

Comparative chromosome painting discloses homologous segments in distantly related mammals.

Comparative chromosome painting, termed ZOO-FISH, using DNA libraries from flow sorted human chromosomes 1, 16, 17 and X, and mouse chromosome 11 discloses the presence of syntenic groups in distantly related mammalian orders ranging from primates (Homo sapiens), rodents (Mus musculus), even-toed ungulates (Muntiacus muntjak vaginalis and Muntiacus reevesi) and whales (Balaenoptera physalus). These mammalian orders have evolved separately for 55-80 million years (Myr). We conclude that ZOO-FISH can be used to generate comparative chromosome maps of a large number of mammalian species.

Animals↗

Chromosome specific paints from a high resolution flow karyotype of the mouse.

Chromosomes from antigen stimulated B-cells from spleens of inbred mice have been separated using flow cytometry into 18 distinguishable peaks. Using locus-specific oligonucleotides and fluorescence in situ hybridization to banded metaphase spreads, 15 individual chromosomes were identified: 1, 2, 3, 6, 7, 8, 9, 11, 12, 16, 17, 18, 19, X and Y. The remaining six chromosomes, occurring as pairs in three peaks, 4 with 5, 10 with 13, and 14 with 15, were resolved by flow sorting chromosomes from mice carrying an appropriate homozygous translocation and 4, 5 and 14 have been isolated in this way. This is the first demonstration of how a complete set of mouse chromosome paints can be produced.

Animals↗

Genetic markers substantiate long-term storage and utilization of sperm by female painted turtles.

Most studies of genetic parentage in natural populations have been limited to a single breeding season or reproductive episode and, thus, provide only a snapshot of individuals' mating behaviours. Female turtles can store viable sperm in their reproductive tracts for as long as several years, but the extent to which this capacity is utilized in nature has remained unknown. Here, we employ microsatellite markers to assess genetic paternity in successive clutches of individually marked, free-ranging female painted turtles (Chrysemys picta) over a four year period. The genetic data from 113 clutches from this natural population demonstrate that most females (80.5%) remated each year and that each female generally used a single male's sperm to fertilize all clutches laid within a year. However, sperm usage among females varied considerably, and some females apparently used sperm that had been stored for up to three years to fertilize some or all eggs laid in consecutive nesting seasons. Thus, remating by females is not necessary for continued offspring production from a given sire. Furthermore, 13.2% of all clutches examined showed evidence of multiple paternity, and the genetic paternity patterns across years suggest a 'last in, first out' operation of the females' sperm storage tubules.

Alleles↗

Ultrastructural changes in granulosa cells and plasma steroid levels after administration of luteinizing hormone-releasing hormone in the Western painted turtle, Chrysemys picta.

In this study we investigated the effects of treatment by luteinizing hormone-releasing hormone (LHRH) on the morphology and steroid release of ovarian tissues in the Western painted turtle, (Chrysemys picta). In Experiment I, four adult female turtles were injected with synthetic mammalian LHRH (i.p., 500 pg/g bodyweight) and four with saline 2-3 weeks prior to ovulation. Granulosa cells from LHRH-treated turtles vs controls contained both preovulatory follicles (16-20 mm in diameter) and small follicles (0.5-1.00mm in diameter) with increased RER, free ribosomes and mitochondria with swollen cristae. An increase in the amount of cytoskeletal material (microfilaments) was observed in granulosa cells of the experimental turtles compared to the controls. Cytoplasmic extensions of the oocyte and granulosa cells were longer in the small follicles of treated animals, accounting for the observed increase in the thickness of the zona pellucida (ZP) over the controls. In Experiment II, administration of LHRH (i.p.) to 10 turtles during the same period triggered a substantial increase in plasma progesterone and estradiol-17beta levels over the 10 saline-injected controls. This supports the idea that in this species, as in mammals, steroidogenic activity in the ovarian follicles are under the control of the hypothalamic-pituitary axis. The ultrastructure and hormonal levels of the experimental animals were typical of untreated turtles just prior to ovulation. In this species the development of follicles and steroidogenesis can be stimulated prematurely by a releasing hormone from a nonreptilian origin.

Animals↗

Modification of a Super-Sep bottle for paint-on application of a separating medium used in flasking dentures.

Modifying a Super-Sep bottle by adding a brush to the lid for paint-on use allows for the precise application of the separating medium used when flasking dentures. Super-Sep offers many advantages over other commonly used separating mediums: It is virtually dimensionless, allowing for better accuracy; it dries quickly and functions best on moist stone; and it can be poured against immediately. The latter fact means less time is needed before the subsequent layers of stone can be applied when denture flasking.

Calcium Sulfate↗

The contribution of colour to three-dimensional ambiguities in paintings and drawings.

This paper is about kinds of visual ambiguity that concern the author as a painter. Some effects of incompatible depth cues on the experience of looking at paintings and drawings are discussed. Particular attention is given to the role of colour as a depth cue. It is suggested that areas of identical pigmentation on different parts of a picture surface tend to be interpreted as being on the same plane. Where this interpretation is incompatible with other depth cues 'visual tension' results, the degree of tension being dependent on the relative strength of the cues involved. This hypothesis is illustrated and elaborated with the help of a number of line drawings.

Art↗

Reconstruction of genomic rearrangements in great apes and gibbons by chromosome painting.

The homology between hylobatid chromosomes and other primates has long remained elusive. We used chromosomal in situ suppression hybridization of all human chromosome-specific DNA libraries to "paint" the chromosomes of primates and establish homologies between the human, great ape (chimpanzee, gorilla, and orangutan), and gibbon karyotypes (Hylobates lar species group, 2n = 44). The hybridization patterns unequivocally demonstrate the high degree of chromosomal homology and synteny of great ape and human chromosomes. Relative to human, no translocations were detected in great apes, except for the well-known fusion-origin of human chromosome 2 and a 5;17 translocation in the gorilla. In contrast, numerous translocations were detected that have led to the massive reorganization of the gibbon karyotype: the 22 autosomal human chromosomes have been divided into 51 elements to compose the 21 gibbon autosomes. Molecular cytogenetics promises to finally allow hylobatids to be integrated into the overall picture of chromosomal evolution in the primates.

Animals↗