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T Pace

Publications and source records attributed to T Pace.

33 records · Page 2Linked to original sources

Organization of subtelomeric repeats in Plasmodium berghei.

Several (but not all) Plasmodium berghei chromosomes bear in the subtelomeric position a cluster of 2.3-kilobase (kb) tandem repeats. The 2.3-kb unit contains 160 base pairs of telomeric sequence. The resulting subtelomeric structure is one in which stretches of telomeric sequences are periodically spaced by a 2.1-kb reiterated sequence. This periodic organization of internal telomeric sequences might be related to chromosome-size polymorphisms involving the loss or addition of subtelomeric 2.3-kb units.

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Presence of contaminating mitochondrial DNA from host reticulocytes in experimental infections of Plasmodium berghei.

Superposition of two unrelated processes, namely terminal reticulocyte differentiation and synchronous plasmodial development, takes place in experimental infections of Plasmodium berghei. The first process is shown to be responsible for the appearance of some discrete restriction bands of host origin when DNA is extracted from leucocyte-free blood containing synchronous parasites at early stages of infection. These discrete DNA fragments cross-hybridize with host cell mitochondrial DNA. Purification steps are suggested to reduce this effect, which might be relevant also in the case of other plasmodial species exhibiting preference for reticulocytes as host cell.

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Telomeric motifs are present in a highly repetitive element in the Plasmodium berghei genome.

Using as probes the subfragments of the telomeric sequence previously cloned by us from Plasmodium berghei DNA, we identified and cloned a 2.3 kb repeat, largely overlapping the original telomeric insert. Restriction mapping indicated that cloned inserts (2.3 kb in length) represented circularly permutated versions of a rather well conserved repeated element, at least in part organized in tandem. The 2.3 kb repeat family with a copy number of about 300 occupies about 4% of the whole genome. The copies are unevenly distributed among the chromosome-sized molecules revealed by pulsed field gradient electrophoresis. Complete sequence determination of the 2.3 kb element revealed that telomere-related motifs are present with a characteristic pattern in a set of tandem repeats, 27 bp long. The perfect conservation of these motifs as well as the pattern of chromosomal distribution suggest that we are dealing with a specialised structure subject to selective mechanisms of amplification and maintenance.

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The relationship between chromosomal origins of replication and the nuclear matrix during the cell cycle.

A cytological investigation into the dynamic behaviour of the origins of replication with respect to the nuclear matrix has been carried out on Xenopus laevis cultured cells. In order to preferentially label origins or 'non-origin' regions along DNA fibres, 5-fluoro-2'-deoxyuridine (FUdR)-treated cells were pulsed with [3H]deoxyadenosine in early or late S phase. Samples were then allowed to proceed through the cell cycle for increasing times. The DNA loops were induced in situ to completely uncoil around the nuclear matrix. The autoradiographic analysis shows that, under the experimental conditions used, 'non-origin' regions behave as expected from previous studies, i.e., they associate with the nuclear matrix only when they become part of a replication fork, whereas active origins of replication remain associated with the matrix throughout the cell cycle.

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Homologous telomeric sequences are present in different species of the genus Plasmodium.

The telomeric sequence cloned from Plasmodium berghei (see M. Ponzi et al. (1985) EMBO J. 4, 2991-2995) was tested for species specificity. A telomeric and a subtelomeric fragment of the cloned insert served as separate, labelled probes on pulsed field gradient electrophoretical patterns and on genomic digests from the rodent malarias Plasmodium yoelii, Plasmodium chabaudi and from the human malaria Plasmodium falciparum. Results indicate that the subtelomeric fragment, abundantly represented in two chromosomes of P. berghei, is not present in the other DNA tested, while the telomeric fragment is present in every chromosome-sized molecule in all the species tested. The telomeric location in the other genomes of the sequences homologous to the P. berghei telomeric probe is confirmed by experiments with Bal 31 exonuclease. In all cases, the TaqI site appears to delimit the common telomeric portion.

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Identification of a telomeric DNA sequence in Plasmodium berghei.

A fragment of Plasmodium berghei DNA was cloned using a technique designed to select for telomeric sequences. The cloned fragment recognizes Bal31-sensitive bands in P. berghei genomic digests. It contains at its distal end at least 70 tandem repeats of the heptanucleotide sequence CCCTGAAA. The presence of natural single strand discontinuities in the telomeric regions of P. berghei DNA is demonstrated by the selective incorporation of deoxyribonucleoside triphosphates in the absence of DNase. The number of copies of the cloned sequence present in each genome agrees with an estimate of 6-12 chromosomes per nucleus.

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Neurobehavioural effects of repeated occupational exposure to toluene and paint solvents.

In studies of two workforces 44 men exposed to paint solvents and 52 men working with toluene were compared with age matched comparison groups of non-exposed workers. Scores on a series of performance tests were examined, the solvent exposed in each study functioning less well than their comparison group. The apparent deficit among the paint exposed workers was substantial, but this was reduced after allowance for estimated intellectual capacity. No deficit was apparent on rematching with a second comparison group of more appropriate measured capacity. No evidence was found of impaired nerve conduction in the ulnar or median nerves, and few clinical signs of neurological damage were apparent. Nevertheless, in an associated inquiry of neuropsychological symptoms among 236 painters and 128 non-exposed workers a highly significant excess of symptoms was found among those exposed to paint solvents.

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Different chromosome banding procedures to distinguish between mouse and rabbit chromosomes in mouse-rabbit somatic cell hybrids.

The discrimination of the parental chromosomes in interspecific somatic cell hybrids, is a powerful tool in gene mapping studies. We report the methods used to distinguish between murine and rabbit chromosomes in mouse-rabbit somatic cell hybrids. By means of G-banding patterns it is possible to determine the murine or rabbit origin of the chromosomes, as well as the single chromosome identification in the same hybrid metaphase. To rapidly establish whether the chromosome are of murine or rabbit origin we use C-banding and Hoechst staining procedures, which staining or elongating respectively and preferentially the centromeric area of the mouse chromosomes allowed indubitable species assignment.

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