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PubMed1998

Microdissection of pig chromosomes: dissection of whole chromosomes, arms and bands for construction of paints and libraries.

Chromosome microdissection is an important means to efficiently generate a large number of markers from a desired region of a genome. The present study was designed to initiate microdissection and amplification of DNA from whole chromosomes, arms, or bands of porcine chromosomes. The following pig (SSC) chromosomes/segments were scraped: SSC1p, SSC1q26-q2.13, SSC2q11-q14, SSC4q12-q25, SSC13, SSC13q12-q31, SSC13q32-q43, SSC13q32-q43, SSC15, and SSC16q21-q23. After amplification and PCR-labelling, the DNA from the dissected segments were painted back to normal metaphase chromosomes to test their identity. Microdissection of some of the segments (on SSC4, 13 and 15) coincides with the mapping of economically important traits. As a first step towards generation of markers, microcloning of amplified product from SSC1p and SSC15 was carried out. The libraries were screened with a (GT)15 oligonucleotide probe. Future prospects of such a work in farm animals are discussed.

Animals↗
PubMed1975

An adhesive paint-on restorative for tooth surface defects.

The physical properties of an adhesive paint-on restorative material have been examined and it is shown that the adhesion to enamel is good, its compressive strength about equal to silicate, hardness is less than for composite resins, and stain resistance is good.

Acrylic Resins↗
PubMed1992

Reverse chromosome painting: a method for the rapid analysis of aberrant chromosomes in clinical cytogenetics.

We describe a method, termed reverse chromosome painting, which allows the rapid analysis of the content and breakpoints of aberrant chromosomes. The method involves the sorting of small numbers of the aberrant chromosome from short term blood culture preparations or cell lines by using bivariate flow karyotype analysis. The sorted chromosomes are amplified and biotin labelled enzymatically using a degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), the product annealed to metaphase spreads from normal subjects, and hybridisation detected using fluorescence in situ hybridisation (FISH). We show the usefulness of this method for routine clinical cytogenetics by the analysis of cases involving an insertion, a deletion, a translocation, and two cases of a chromosome with additional material of unknown origin. The method has particular application for the rapid resolution of the origin of de novo unbalanced chromosome duplications.

Adult↗
PubMed1994

Pseudodicentric chromosome 18 diagnosed by chromosome painting and primed in situ labelling (PRINS).

We report on a newborn white male infant with marked dysmorphic features and various congenital malformations. The initial clinical evaluation showed Crouzon-like features as well as some features of trisomy 18 syndrome and trisomy 13 syndrome. The results from conventional cytogenetic analysis showed a structurally abnormal chromosome replacing one normal chromosome 18, but only by applying molecular cytogenetic methods could the architecture of this abnormal chromosome be characterised clearly. The primed in situ labelling (PRINS) technique, using a newly synthesised alpha 18 oligonucleotide, showed the dicentric pattern and direct chromosome painting established the origin to be from chromosome 18. The combination of conventional cytogenetics and molecular cytogenetics showed the karyotype in the proband to be 45,XY,-14,-18,-21,+t(14;21),+psu dic(18) (qter-->cen-->p11.3: :p11.3-->psu cen-->qter). This was supported by molecular analysis using chromosome 18 specific DNA markers, which showed the paternal origin of the abnormal chromosome.

Abnormalities, Multiple↗
PubMed1994

Elucidation of structural abnormalities of the X chromosome using fluorescence in situ hybridisation with a Y chromosome painting probe.

Particular regions of the X and Y chromosomes share DNA sequence homology to the extent that cross hybridisation occurs. Thus, chromosome painting with a whole Y chromosome probe consistently results in fluorescence on specific regions of the X chromosome as well as the complete Y chromosome. This phenomenon has been exploited to elucidate the structure of unusual X chromosome rearrangements, without Y involvement, in two females.

Adult↗
PubMed1992

Natural freezing survival by painted turtles Chrysemys picta marginata and C. picta bellii.

Hatchlings of both the Midland (Chrysemys picta marginata) and Western (C. picta bellii) subspecies of the painted turtle tolerate the freezing of extracellular body fluids while overwintering in terrestrial nests. Fall-collected hatchlings survived 3 days of continuous freezing at -2.5 degrees C, with ice contents of 43.5 +/- 1.0% of total body water (SE; n = 24) for C. picta marginata and 46.5 +/- 0.8% (n = 32) for C. picta bellii. Survival times dropped to 4-5 h when temperature was lowered to -4 degrees C, correlated with ice contents of greater than or equal to 50%. However, C. picta marginata tested immediately after excavation from nests in the spring showed greater freeze tolerance, with survival extending to 11 days at -2.5 degrees C and a higher mean ice content of 50.2 +/- 1.2% (n = 6). Spring hatchlings also had high supercooling points, -1.07 +/- 0.13 degrees C (n = 8), that dropped within 3 days to -4.83 +/- 0.83 degrees C (n = 4), suggesting a breakdown of endogenous ice-nucleating agents when hibernation ended. A search for possible cryoprotectants showed that both subspecies accumulated glucose and lactate in liver during freezing (net increase = 3-13 mumols/g wet wt); both also maintained large free amino acid pools in organs, with taurine making up 21-47% of the total.

Acclimatization↗
PubMed1998

Sperm segregation analysis of a complex chromosome rearrangement, 2;22;11, by whole chromosome painting.

Using the human sperm-hamster oocyte fusion technique and whole chromosome painting, we studied sperm chromosome segregation in a male heterozygous for a complex chromosome rearrangement, 46,XY,-2, +der(2)t(2;11)(q13; q23),-11,+der(11)t(11;22)(q23;q11.2),-22, +der(22)t(2;22)(q13; q11.2). A total of 208 sperm complements were analyzed. The frequency of sperm carrying a normal or a balanced complement was 13.5% (9.62% and 3.85%, respectively). The frequency of unbalanced sperm was 86.5% (64.9% from 3:3 segregation, including 30 different types; 20.7% from 4:2 segregation, including 21 different types; and 0.96% from 5:1 segregation, including 2 different types). The sex ratio, determined in 134 sperm complements, did not differ from the expected 1:1 ratio. The results obtained in this study are compatible with the formation, during the synaptic process, of a complex hexavalent figure involving chromosomes 2, 11, and 22. The behavior and segregation of this complex figure would explain the high frequency (86.5%) of unbalanced complements observed in this carrier.

Animals↗
PubMed1993

Translocations between two specific human chromosomes detected by three-color "chromosome painting".

Translocations between two specific chromosomes are important markers for many human malignancies. Previously, the detection of translocations involving random breakpoints between two specific chromosomes could only be accomplished by banding techniques, which are severely labor intensive and require highly trained technicians. The three-color chromosome painting approach described in this paper was developed in our laboratory to detect translocations between two specific human chromosomes rapidly and accurately, while simultaneously revealing the nonhybridized chromosomes. Because this method efficiently detects translocations involving breakpoints anywhere on the targeted chromosomes, it is ideal as a screening tool for chromosome-specific translocations.

Chromosomes, Human↗
PubMed1994

Isolation of rat chromosome-specific paint probes by bivariate flow sorting followed by degenerate oligonucleotide primed-PCR.

The flow karyotype of rat chromosomes was determined by dual beam flow cytometry. Eighteen fractions were sorted and subsequently amplified by degenerate oligonucleotide primed-PCR as described by Telenius et al. (1992a, 1992b). The PCR products were labeled and used as probes for fluorescence in situ hybridization on rat fibroblast metaphases. The amplified chromosomes were detectable as bright chromosome paints and in most cases the signal was evenly distributed along the whole chromosome except for the centromeric region in half of the chromosomes. The results show that chromosomes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 19, 20, X and Y can be sorted as individual fractions, whereas chromosomes 11, 13, 14, 15 and chromosomes 16, 17 and 18 are clustered together in the flow karyotype.

Animals↗
PubMed1995

A panel of subchromosomal painting libraries representing over 300 regions of the human genome.

DNA samples from about 100 human-hamster somatic cell hybrids, previously characterized by conventional banding techniques, were amplified with dual-Alu PCR. The products were then used as probes in FISH experiments on normal human metaphases for an accurate cytogenetic characterization of the human material retained in each hybrid. In addition to entire chromosomes, most hybrids were found to contain one or a few chromosome fragments, as a result of rearrangements that had occurred in vitro. Forty additional primary hybrids, in which conventional cytogenetic analysis failed to reveal any complete human chromosome, contained many human chromosome fragments. More than 300 chromosome fragments were scored and their precise chromosomal location recorded. We show data indicating that subchromosomal painting libraries generated from these hybrids can be favorably used in the fine characterization of chromosomal rearrangements encountered in clinical cytogenetics or in tumor cytogenetics, and in tracking chromosomal changes that occurred in primate evolution.

Animals↗
PubMed1995

Chromosome painting analysis of early oogenesis in human trisomy 18.

We have used chromosome 18-specific painting probes to analyze early stages of oogenesis in two human trisomy 18 fetuses. At leptotene, a diffuse, nonlinear chromosomal fluorescence was detected as one (27%), two (42%), or three (31%) signals in 534 cells. The variation in size of these signals implies the possibility of associations between homologs prior to zygotene. At pachytene, about 75% (339/453) of the cells had a trivalent configuration, and almost half of these cells exhibited almost complete triple synapses. Approximately 24% of the pachytene cells demonstrated a bivalent:univalent configuration, and 1% exhibited complete asynapsis. Our data imply that triple synapses may be a regular feature of meiosis involving multivalents.

Abortion, Induced↗
PubMed1995

Chromosome painting with human chromosome-specific DNA libraries reveals the extent and distribution of conserved segments in bovine chromosomes.

Commercially available human chromosome-specific DNA libraries covering the whole karyotype were hybridized to normal bovine metaphase spreads to characterize the conserved chromosomal segments between man and cattle. All chromosome libraries except the Y chromosome library displayed a signal on at least part of one or more bovine chromosomes. The labeling was clearly visualized and permitted precise delineation of the hybridized bovine chromosomal segments. This study indicates that the reorganization of the genetic material between human and bovine genomes is not as great as expected from classical comparative cytogenetics based on banding patterns. However, apart from interchromosomal rearrangements between ancestral forms of human and bovine chromosomes, a majority of intrachromosomal rearrangements must have occurred in these species during evolution to explain the differences in the banding patterns of their chromosomes. These results show that chromosome painting with heterologous chromosome-specific DNA libraries can provide useful information in comparative studies on karyotypes and gene maps of distantly related mammalian species. The observations are discussed in relation to published data on gene mapping in man and cattle.

Animals↗
PubMed1996

Demonstration of homoeologies between human and lemur chromosomes by chromosome painting.

Human-specific probes for chromosomes 3, 7, 9, 14, 19, and 21 were used to paint chromosomes of three lemur species: Eulemur fulvus mayottensis, E. macaco macaco, and Lemur catta. Chromosomes 1 and 3 of E. f. mayottensis are homoeologous to human chromosomes 3, 9, 14, and 21, as previously suggested by chromosome banding. Probes for human chromosomes 7 and 19 produced unexpectedly strong signals in the centromeric regions of all lemur chromosomes, suggesting that sequences homologous to nonrepeated sequences of the human genome have been amplified during the formation of constitutive heterochromatin in lemurs.

Animals↗
PubMed1996

Chromosome homology between the domestic pig and the babirusa (family Suidae) elucidated with the use of porcine painting probes.

Homology among three pairs of domestic pig (Sus scrofa) and five pairs of babirusa (Babyrousa babyrussa) autosomes has been demonstrated with the use of porcine painting probes. With the results of this study, in addition to data obtained earlier through the application of banding techniques, correspondence between all individual chromosomes of these two distantly related pigs has been identified.

Animals↗
PubMed1994

Diuretic plants in the paintings of Pompeii.

The plants that appear in the paintings and mosaics of Pompeii are chiefly edible and medicinal, though flowers with purely esthetic appeal are also shown. An important example is one of the floor mosaics from the House of the Faun, in which it is possible to identify lemon, cherry, strawberry, pomegranate, grape and olive, leaves of grape, fig, apple and olive, and flowers of corn cockle. The diuretic properties of some of these plants are mentioned in the Naturalis Historia of Pliny the Elder. A silver cup from the House of Menandro, one of the most refined examples of plant decoration in antiquity, shows olive branches and fruits. The presence of plants in such artefacts confirms that people of classical times were conscious that plants were important producers of food, oils, fibers, woods and medicines.

Diuretics↗
PubMed1990

Correlation between lymphocyte proliferative responses and dendritic cell migration in regional lymph nodes following skin painting with contact-sensitizing agents.

We have investigated whether there exists a correlation between the induction of draining lymph node cell (LNC) proliferation in contact allergy and the accumulation of dendritic cells (DC) within such lymph nodes. CBA/Ca mice, which compared with mice of BALB/c strain, mount a more vigorous lymphocyte proliferative response following sensitization, also exhibited a more marked accumulation of DC in draining lymph nodes 24 h following skin painting. Moreover, studies with the skin-sensitizing fluorochromes fluorescein isothiocyanate (FITC) and rhodamine B isothiocyanate (RITC) revealed that DC-enriched fractions of draining LNC prepared from CBA/Ca mice contained a higher percentage of antigen-bearing cells than did those from BALB/c mice. A relationship between DC migration into lymph nodes and the magnitude of the induced LNC proliferative response was also indicated by experiments performed in BALB/c mice with a variety of contact allergens. It was observed that there was a direct correlation between the vigour of the proliferative response measured 3 days following exposure and the frequency of DC in draining nodes at 24 h. Collectively these data suggest that following skin sensitization the migration of DC into the draining lymph nodes influences quantitatively the primary immune response and the development of contact allergy.

Allergens↗
PubMed1986

Distribution of 2,4-dinitrophenyl groups on the epidermal Langerhans cells of guinea pigs following skin painting with 2,4-dinitrochlorobenzene.

It has been postulated that epidermal Langerhans cells (LC) are essential for induction and elicitation of contact sensitivity (CS) to simple chemical allergens. Although the precise role of LC in CS has not been determined, recent reports have focused on a function for LC in the presentation of antigen to T lymphocytes. To investigate this possibility, the distribution of allergen on the epidermal LC of inbred strain 13 guinea pigs was observed by the immunofluorescent method using antibodies against Ia antigen and hapten. 2,4-Dinitrophenyl groups were found to be localized on/in Ia-positive epidermal LC of the animals following skin painting with 2,4-dinitrochlorobenzene. This finding is discussed in relation to the role of LC as antigen presenting cells in CS.

2,4-Dinitrophenol↗
PubMed1976

Experimental lead paint poisoning in nonhuman primates. I. Clinical signs and course.

Lead-containing paints were administered orally to 27 rhesus monkeys for periods of 18-667 days. Lead acetate was fed to nine monkeys of three different species for 9-156 days. Excretion of one week's dose of lead in six primates ranged from 35 to 94%. The animals incurred moderate to extreme elevations of lead in blood, most lost weight, or had depressed weight gains, and developed Burtonian lines, some died suddenly and unexpectedly, and many terminated in a moribund state with profound anemia. Only one neonate had obvious signs of lead encephalopathy. The monkeys' ages, dose and source of lead, and possibly other factors, affected their response to lead.

Age Factors↗