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Introgression of Lilium rubellum Baker chromosomes into L. longiflorum Thunb.: a genome painting study of the F1 hybrid, BC1 and BC2 progenies.

Interspecific hybrids between Lilium longiflorum (L, 2n = 2x = 24) and Lilium rubellum (R, 2n = 2x = 24) were produced with the aim of transferring desirable horticultural traits from L. rubellum to L. longiflorum. All F1 hybrids (LR, 2n = 2x = 24) and BC1 individuals (LLR, 2n = 3x = 36) were phenotypically uniform for plant height, flowering time, leaf shape and flower colour. The BC1 plants were, in spite of their triploid nature, fertile and could be used as a female parent in backcrossings with autotetraploid L. longiflorum (LLLL, 2n = 4x = 48). Twelve BC2 individuals were obtained and three of them were selected for further chromosome analysis. As L. longiflorum and L. rubellum chromosomes were indistinguishable in the hybrids, genomic in-situ hybridization (GISH) was applied to establish the parentage of the chromosomes of the F1 hybrids and the BC1 and BC2 progenies. GISH confirmed the LLRR constitution of the doubled amphimonoploid (allodiploid), and the LLR constitution of all BC1 plants. The three selected BC2 plants were, as expected, aneuploid, containing three complete sets of L. longiflorum chromosomes and six, seven or eight L. rubellum chromosomes, respectively. However, L/R translocation or recombinant chromosomes could not be demonstrated in the mitotic metaphase complements of the F1, BC1 and BC2 plants. In spite of the high frequencies of homoeologous recombination in the F1 hybrids (LR) pollen was found to be sterile in all cases. At metaphase I of the pollen mother cells of the BC1 plants, genome painting did not reveal any cases of homoeologous pairing and recombination between L and R chromosomes. This lack of exchange between homoeologous chromosome segments indicates complete preferential pairing of the L and R chromosomes in the F1 (amphidiploid) and BC1 plants. It seems that the preferential pairing in the F1 and BC1 hybrids hinder the introgression of the chromosome segments or species-specific genes into the recipient for breeding purposes.

Chimera↗

Chromosome painting in marsupials: genome conservation in the kangaroo family.

In order to deduce the ancestral genome arrangement in the karyotypically diverse marsupial family Macropodidae, and to assess chromosome change in this family, chromosome-specific paints from the tammar wallaby (2n = 16) were hybridized to metaphase spreads from the two species proposed to represent the 2n = 22 ancestral karyotype, as well as species with derived 2n = 20 and 2n = 14 karyotypes. Identical patterns were observed in the two 2n = 22 species, from which the rearrangements to form the three derived karyotypes may be easily deduced to be 1, 3 and 4 different fusions, respectively. The identical Thylogale and Dorcopsis genomes may both be used to represent the pleisiomorphic macropodid chromosome complement. Variation in the X chromosome was also investigated by hybridizing an X-Y shared tammar wallaby 12-kb repeat element to chromosomes from the other four macropodid species, finding that it hybridized only to the most closely related species, and therefore is of recent origin.

Animals↗

Chromosome painting supports lack of homology among sex chromosomes in Oncorhynchus, Salmo, and Salvelinus (Salmonidae).

The sex chromosome pair has been identified previously as the largest submetacentric pair in the genome in several species of the genus Salvelinus (eastern trouts and chars) including S. namaycush (lake trout) and as a large subtelocentric/acrocentric pair in several species of the genus Oncorhynchus (Pacific trouts and salmon). Sex chromosomes have not been identified in Salmo (Atlantic salmon and brown trout). Two paint probes, one specific for the short arm (Yp) and the other for the long arm (Yq) of the sex chromosome pair in Salvelinus namaycush were hybridized to chromosomes of Oncorhynchus mykiss (rainbow trout) and O. tshawytscha (chinook salmon) and Salmo salar (Atlantic salmon) and S. trutta (brown trout). The two probes hybridized to two different autosomal pairs in each of the Oncorhynchus species, supporting lack of homology between the sex chromosomes in the two genera. The Yp probe hybridized to interstitial regions on two different chromosome pairs in S. salar and one pair in S. trutta. The Yq probe hybridized to a different pair in both species.

Animals↗

Painting analysis of meiotic metaphase I configurations of the germ line-limited chromosomes in Acricotopus.

Meiotic metaphase I configurations and pairing behavior of the germ line-limited chromosomes (= Ks) in the chironomid Acricotopus lucidus were analyzed by chromosome painting using specific probes of the three soma chromosomes (= Ss) and of their individual arms. The Ks are derived from the Ss and possess large S-homologous sections. Beside regular K and S bivalents, we also observed frequently K multivalents, e.g. trivalents, mainly quadrivalents, but also penta- and hexavalents, composed of the same K type in metaphases I. Chiasmata predominately occur within the S-homologous sections, probably ensuring a correct segregation and the transmission of a set of Ks to the next generation. Because K bivalents are almost exclusively autobivalents in A. lucidus formed by earlier sister chromatids, this multivalent formation with crossover also between homologous but non-identical Ks leads to genetic recombination within a K type. Rarely, quadrivalents composed of non-homologous Ks but derived from the same S were found. Therefore, these multivalents most probably resulted from crossover between homologous sections of morphologically different K types. This may result in new K types and might be important for the evolution of K type diversity in A. lucidus. In some cases, pairing-like associations between SIII and K4, which is derived from SIII, were observed in metaphases, indicating the possibility of crossover events and recombination between these chromosomes and so between the somatic and the germ-line restricted chromosome complements. Possible functions of additional copies of S sequences carried in the germ line are discussed.

Animals↗

Chromosomal painting detects non-random chromosome arrangement in dasyurid marsupial sperm.

Chromosome arrangements have been studied in metaphase and interphase somatic cells and in sperm of many animal species, but there are conflicting data and it is still not clear whether chromosomes are arranged randomly or non-randomly. We used chromosome painting to reveal the positions of chromosomes in marsupial sperm. Marsupials are ideally suited for these studies because they have only a few large chromosomes. Here, we show that chromosomes occupy fixed positions in the immature and mature sperm of Sminthopsis crassicaudata. We suggest that the non-random arrangement of chromosomes in marsupial sperm may be important in establishing chromosome arrangement and patterns of gene activity within the developing embryo.

Animals↗

The phylogeny of howler monkeys (Alouatta, Platyrrhini): reconstruction by multicolor cross-species chromosome painting.

We performed multidirectional chromosome painting in a comparative cytogenetic study of the three howler monkey species Alouatta fusca, A. caraya and A. seniculus macconnelli (Atelinae, Platyrrhini) in order to reconstruct phylogenetic relationships within this genus. Comparative genome maps between these species were established by multicolor fluorescence in-situ hybridization (FISH) employing human, Saguinus oedipus and Lagothrix lagothricha chromosome-specific probes. The three species included in this study and previously analyzed howler monkey species were subjected to a phylogenetic analysis on the basis of a data matrix comprised of 98 discrete molecular cytogenetic characters. The results revealed that howler monkeys represent the genus with the most extensive karyotype diversity within Platyrrhini so far analyzed with high levels of intraspecific chromosomal variability. Two different multiple sex chromosome systems were identified. The phylogenetic analysis indicated that Alouatta is a monophyletic clade which can be derived from a proposed ancestral Atelinae karyotype of 2n = 62 chromosomes by a chromosome fusion, a fission, a Y-autosomal translocation and a pericentric inversion. Following these suggestions, the genus Alouatta can be divided into two distinct species groups: the first includes A. caraya and A. belzebul, the second A. s. macconnelli, A. sara, A. s. arctoidea and A. fusca.

Alouatta↗

Chromosomal studies in Callicebus donacophilus pallescens, with classic and molecular cytogenetic approaches: multicolour FISH using human and Saguinus oedipus painting probes.

This paper presents the karyotype of Callicebus donacophilus pallescens for the first time. The analysis included G-, C-, NOR-banding techniques and FISH with chromosome painting probes from Saguinus oedipus and Homo sapiens. The results were compared with the karyotypes of Callicebus moloch donacophilus and C. moloch previously published. These three karyotypes display the same diploid number (2n = 50) but diverge about the number of biarmed and acrocentric chromosomes. The acrocentrics 14 and 15 from C. m. donacophilus and C. moloch have undergone an in-tandem fusion originating a large acrocentric (pair 10) in C. d. pallescens. The major submetacentric pair (pair 1) from C. d. donacophilus and C. moloch have undergone fission originating two acrocentric pairs in C. d. pallescens (pairs 15 and 22). Herein was evidence that, in spite of the high interspecific variation among Callicebus, most of the chromosomes remained conserved.

Animals↗

Precise identification of chicken chromosomes in the lampbrush form using chromosome painting probes.

Chromosome painting probes specific for macrochromosomes 1, 2, 3, 4, 5, and Z were applied to both mitotic and lampbrush chromosomes of the chicken (Gallus gallus domesticus). Five autosomal macrobivalents and sex chromosome Z in the lampbrush phase were identified and their correspondence to the target chromosomes in the metaphase of mitosis was shown. Nascent transcripts on lateral loops of the target lampbrush chromosome were intensively labelled when the hybridization was performed without RNase A treatment according to the DNA/(DNA + RNA) hybridization protocol.

Animals↗

Refined genome-wide comparative map of the domestic horse, donkey and human based on cross-species chromosome painting: insight into the occasional fertility of mules.

We have made a complete set of painting probes for the domestic horse by degenerate oligonucleotide-primed PCR amplification of flow-sorted horse chromosomes. The horse probes, together with a full set of those available for human, were hybridized onto metaphase chromosomes of human, horse and mule. Based on the hybridization results, we have generated genome-wide comparative chromosome maps involving the domestic horse, donkey and human. These maps define the overall distribution and boundaries of evolutionarily conserved chromosomal segments in the three genomes. Our results shed further light on the karyotypic relationships among these species and, in particular, the chromosomal rearrangements that underlie hybrid sterility and the occasional fertility of mules.

Animals↗

Generation of chromosome painting probes from single chromosomes by laser microdissection and linker-adaptor PCR.

Fluorescence in situ hybridization (FISH) plays an essential role in research and clinical diagnostics. The versatility and resolution of FISH depends critically on the probe set used. Here, we describe an improved approach for the generation of specific DNA probes from single copies of chromosomes. Single chromosomes or single chromosomal regions were microdissected by laser pressure catapulting and amplified using linker-adaptor PCR. The probes were labeled and tested in various scenarios including multicolor-FISH experiments employing up to seven different fluorochromes. FISH confirmed the specific and even staining of the respective chromosomal regions. Furthermore, the capability of these probes to detect even small translocations (<3 Mb) suggests that the dissected regions are completely represented in the generated painting probes.

Chromosome Painting↗

Airborne allergic contact dermatitis from 3-iodo-2-propynyl-butylcarbamate at a paint factory.

3-Iodo-2-propynyl-butylcarbamate (IPBC) is a fungicide used in both industrial products and cosmetics. We report the first case of allergic contact dermatitis from airborne exposure to this preservative. A 34-year-old female production worker at a paint factory developed dermatitis on air-exposed skin areas. Patch testing showed a ++ reaction to the preservative IPBC 0.01% in petrolatum. The compound was used as a preservative in wood treatment products manufactured at her work place. Based on animal studies, IPBC is considered safe as a cosmetic preservative. However, widespread use of the chemical might lead to increasing levels of contact allergy, and therefore, close monitoring of IPBC is recommended.

Adult↗

Palaeolithic paintings. Evolution of prehistoric cave art.

Sophisticated examples of European palaeolithic parietal art can be seen in the caves of Altamira, Lascaux and Niaux near the Pyrenees, which date to the Magdalenian period (12,000-17,000 years ago), but paintings of comparable skill and complexity were created much earlier, some possibly more than 30,000 years ago. We have derived new radiocarbon dates for the drawings that decorate the Chauvet cave in Vallon-Pont-d'Arc, Ardèche, France, which confirm that even 30,000 years ago Aurignacian artists, already known as accomplished carvers, could create masterpieces comparable to the best Magdalenian art. Prehistorians, who have traditionally interpreted the evolution of prehistoric art as a steady progression from simple to more complex representations, may have to reconsider existing theories of the origins of art.

Archaeology↗

J. M. W. Turner's painting "The unpaid bill, or the dentist reproving his son's prodigality".

In November 2002, the BDA News carried an item, illustrated with a colour reproduction, describing a painting of a Georgian dentist's rooms by Joseph Mallord William Turner (1775-1851), one of the most respected of English artists, which was shortly to come up for auction at Christies' Rooms in London. This work, first exhibited in 1808, was entitled "The unpaid bill, or the dentist reproving his son's prodigality", and had originally been commissioned by the connoisseur Richard Payne Knight (1750-1824). "The examiner", a contemporary London journal, identifies the 'cradle-piece' for the commission as being a Rembrandt which Payne Knight owned, and the journalist Robert Hunt said that Turner had more than come up to the task of showing that a modern could handle light as well as the old master, 'for a picture of colouring and effect, it is ... inestimable'.

Dental Offices↗

Evaluation of tryptamine in an impinger and on XAD-2 for the determination of hexamethylene-based isocyanates in spray-painting operations.

Tryptamine was evaluated as a reagent for derivatizing hexamethylene diisocyanate (HDI) monomer and oligomers during actual spray-painting operations. In one side-by-side sampling study, an impinger filled with 1-(2-methoxyphenyl)piperazine in toluene was compared with a second impinger filled with tryptamine in dimethyl sulfoxide (DMSO). The amount of HDI monomer obtained was below the limit of quantification for both impingers. The amount of HDI oligomer obtained when using 1-(2-methoxyphenyl)piperazine in toluene was comparable to the amount obtained when using an impinger filled with tryptamine in DMSO. In a second side-by-side sampling study, a tryptamine-coated XAD-2 resin was used as a sorbent. The relative collection efficiency of the tryptamine-coated XAD-2 resin was on average 60% of the value obtained using an impinger filled with tryptamine in DMSO. The results indicate that using an impinger filled with tryptamine in DMSO gives higher concentrations of isocyanate than a tryptamine-coated XAD-2 sorbent for HDI monomer and oligomer.

Dimethyl Sulfoxide↗

Determination of hexamethylene diisocyanate-based isocyanates in spray-painting operations. Part 2. Comparison of high performance liquid chromatography with capillary zone electrophoresis.

A capillary zone electrophoresis (CZE) approach was developed for the determination of hexamethylene diisocyanate (HDI) monomer and HDI-based oligomers. A comparison of CZE with high performance liquid chromatography (HPLC) indicates that the CZE separation completely isolates isocyanates from excess solvent, derivatizing reagent and pigment while offering a fivefold increase in sensitivity. The CZE approach allows for the quantification of HDI monomer and oligomer within a 1 min time window under the run conditions selected. For the determination of HDI-based oligomer, provided that the relative response with respect to HDI monomer is calculated, there is no significant difference (p < 0.05, n = 10) in the isocyanate air concentration when using either HPLC or CZE. The results are significant because they indicate that CZE has advantages for the determination of both HDI-based oligomer and HDI monomer generated during spray-painting operations.

Air Pollutants, Occupational↗

Bacterial chromosomal painting for in situ monitoring of cultured marine bacteria.

We previously described a new method, bacterial chromosomal painting (BCP), for the in situ identification of bacterial cells. Here, we describe the application of this technique to study the ecology and physiology of cultured marine pelagic bacteria from the western Sargasso Sea (WSS). A total of 86 bacteria were isolated from seawater collected from near the surface, at a depth of 250 m and from nutrient-amended seawater incubations. The 10 bacterial isolates that were best represented in environmental genomic DNA from the WSS were selected using reverse genome probing. BCP hybridization cell counts were used to determine the depth-specific distribution of one of the alpha proteobacterial isolates, B5-6, in the WSS during two thermal stratification regimes: stratified and partially mixed. The maximum cell count measured for B5-6 at the summer deep chlorophyll maximum was approximately 4% of the total cell count. This study is the first application of BCP to natural environments.

Alphaproteobacteria↗

Carbamazepine, hepatotoxicity, organic solvents, and paints.

Hepatotoxicity secondary to carbamazepine is a serious condition which can be fatal. However, other concomitant medications or environmental factors may be the offending agents. In this case report, hepatotoxicity secondary to organic solvents and paints is described.

Alcohol Drinking↗