Prospective study of beverage use and the risk of kidney stones

We have analysed the replication of the heterochromatic megachromosome that was formed de novo by a large-scale amplification process initiated in the centromeric region of mouse chromosome 7. The megachromosome is organized into amplicons approximately 30 Mb in size, and each amplicon consists of two large inverted repeats delimited by a primary replication initiation site. Our results suggest that these segments represent a higher order replication unit (megareplicon) of the centromeric region of mouse chromosomes. Analysis of the replication of the megareplicons indicates that the pericentric heterochromatin and the centromere of mouse chromosomes begin to replicate early, and that their replication continues through approximately three-quarters of the S-phase. We suggest that a replication-directed mechanism may account for the initiation of large-scale amplification in the centromeric regions of mouse chromosomes, and may also explain the formation of new, stable chromosome segments and chromosomes.     

Amplification of telomeric DNA and the extent of karyotypic evolution

The distribution of telomeric DNA in the genomes of the antelope ground squirrel, Ammospermophilus harrisii (family Sciuridae; 2n = 32) and the African black-footed cat, Felis nigripes (family Felidae; 2n = 38) were compared by fluorescence in situ hybridization (FISH) technique. These two mammalian species have the highest and the lowest amount of C-banded regions, respectively. FISH preparations with the human telomeric DNA probe showed that all C-banded segments in the A. harrisii chromosomes, except a few intercalary segments, were hybridizing with this DNA. F. nigripes showed hybridization only on the termini of each chromosome, and the C-banded regions did not hybridize with telomeric DNA on FISH analysis. The C-banded chromosomal arms in another rodent species, Peromyscus eremicus (family Cricetidae; 2n = 48), when hybridized with human telomeric DNA showed signals only in the termini of chromosomes but not in the heterochromatic arms. These observations indicate that not all C-banded regions in rodent species are telomeric DNA. The amplification of telomeric DNA in relation to speciation is discussed.     

The relationship of a specific chromosomal region to the development of the acrosome

In early spermatids of Urodeles the chromosome segments bearing constitutive heterochromatin are localized in one half of the round nucleus; this region becomes the basal part of the long nucleus of the spermatozoon. The euchromatic chromosome segments extend toward the anterior nuclear pole in a bouquet configuration (Macgregor and Walker, 1973). In the course of spermiohistogenesis, one of the heterochromatic regions (the acrosomal chromocenter) migrates from the basal part to the anterior half of the spermatid nucleus. This heterochromatic block is identical with a species-specific, definite C-band in the karyotype. This relationship between the acrosomal chromocenter and a specific chromosomal C-band was established in Triturus cristatus, T. marmoratus, T. alpestric and Cynops pyrrhogaster. In closely related species this particular C-band lies on similar chromosomes. - While the spermatid nucleus still retains its round shape the acrosomal chromocenter despiralizes into a long heterochromatic thread (acrosomal thread). Precisely at the position of this thread the nucleus evaginates and acquires a pear-like shape. During the elongation of the nuclear protrusion the acrosomal thread remains associated with the anterior end. At termination of spermiogenesis it lies closely below the acrosome in the tip of the spermatozoon. Spontaneous aberrations which affect the acrosomal chromocenter or the thread lead to the development of spermatozoa with defective tips. - Several euchromatic segments, interspersed between the heterchromatic segments, can be recognized in the completely despiralized acrosomal thread. Genes responsible for the morphogenetic activities of both, the acrosomal chromocenter and the acrosomal thread, in the development of the spermtip, might be localized in these interspersed euchromatic segments. The existence in higher vertebrates of an acrosomal chromocenter or an equivalent chromosomal region is discussed.     

ZOO-FISH and R-banding reveal extensive conservation of human chromosome regions in euchromatic regions of river buffalo chromosomes

Commercially available human chromosome (HSA) painting probes were hybridized on river buffalo (Bubalus bubalis, 2n = 50) chromosomes by using FISH and R-banding techniques. Clear hybridization FITC-signals revealed extensive conservation of human chromosome regions in this species and demonstrated that human chromosome probes primarily paint euchromatic regions (R-bands). The present results are discussed in the light of previous gene mapping data obtained in river buffalo and ZOO-FISH data in cattle, and in relation to the standard bovine chromosome nomenclatures. In particular, HSA 8, HSA 10, HSA 11, and HSA 16+7 paint, respectively, BBU 1p, BBU 4p, BBU 5p, and BBU 24, which are homoeologous, respectively, to cattle chromosomes 25, 28, 29 and 27. Thus, these river buffalo chromosome arms can serve as markers to resolve discrepancies in the nomenclature of cattle and related species.     

Hypomethylation of human sperm pronuclear chromosomes

Using the methylase SssI enzyme, we have analyzed the degree of in situ methylation of human sperm pronuclear chromosomes obtained by fertilizing hamster oocytes with human sperm. Untreated (control) sperm chromosome complements showed a higher degree of in situ methylation, compared to sperm complements previously treated with 5-azadeoxycytidine or lymphocyte chromosomes. This indicates that human sperm pronuclear chromosomes have a lower degree of genomic methylation compared to that of other somatic cells. The similarity in the degree of in situ methylation of the euchromatic and heterochromatic regions of chromosomes 1, 9, 15, and 16 and the Y chromosome in human sperm does not support the existence of a possible correlation between hypomethylation and heterochromatin decondensation.     

Specific interactions of chromatin with the nuclear envelope: positional determination within the nucleus in Drosophila melanogaster

Specific interactions of chromatin with the nuclear envelope (NE) in early embryos of Drosophila melanogaster have been mapped and analyzed. Using fluorescence in situ hybridization, the three-dimensional positions of 42 DNA probes, primarily to chromosome 2L, have been mapped in nuclei of intact Drosophila embryos, revealing five euchromatic and two heterochromatic regions associated with the NE. These results predict that there are approximately 15 NE contacts per chromosome arm, which delimit large chromatin loops of approximately 1-2 Mb. These NE association sites do not strictly correlate with scaffold-attachment regions, heterochromatin, or binding sites of known chromatin proteins. Pairs of neighboring probes surrounding one NE association site were used to delimit the NE association site more precisely, suggesting that peripheral localization of a large stretch of chromatin is likely to result from NE association at a single discrete site. These NE interactions are not established until after telophase, by which time the nuclear envelope has reassembled around the chromosomes, and they are thus unlikely to be involved in binding of NE vesicles to chromosomes following mitosis. Analysis of positions of these probes also reveals that the interphase nucleus is strongly polarized in a Rabl configuration which, together with specific targeting to the NE or to the nuclear interior, results in each locus occupying a highly determined position within the nucleus.     

Treatment of temporomandibular disorders: arthroscopy, orthodontics, kinesitherapy]

In(1LR)pn2a is a pericentric inversion with a euchromatic breakpoint in the 2E polytene region and a heterochromatic breakpoint in the right arm of the X chromosome. It is associated with position-effect variegation (PEV) of the pn, wapl, Pgd and other vital loci of the 2E region, which are relocated near the bulk of the X heterochromatin. Cytological analysis showed that the rearrangement brings the 1A-2E euchromatic segment directly into contact with a major portion of the h34 block, a heterochromatic region that is positively stained by the N-banding technique and contains the AAGAG satellite sequences. Molecular cloning revealed the presence of a new junction between euchromatin and AAGAG satellite sequences and demonstrated that the euchromatic breakpoint of In(1LR)pn2a lies in the vinculin gene. In the X ray-induced secondary rearrangement In(1LR)r30, consisting of a pericentric inversion superimposed on In(1LR)pn2a, the h34 material remains associated with the 2E region but is separated from the rest of the X heterochromatin. In this case, the pn, wapl and Pgd loci no longer variegate, suggesting that the satellite-containing h34 region is not able per se to induce detectable PEV on the adjacent euchromatic genes.     

Orientation of interphase chromosomes as detected by Giemsa C-bands

The orientation of Giemsa C-bands has been studied in mitotic and interphase cells of Allium cepa. A sativum and of Aloe vera. The C-bands in these three species are located at the telomeres, secondary constriction region of the nucleolar chromosomes and the centromeric regions, respectively. Observations in A. cepa and Aloe indicate clearly that the interphase chromosomes are non-random in their orientation and possibly maintain their telophase configuration through the attachment of telomeres and perhaps of kinetochores with the nuclear membrane. Electron micrographs of onion cells also reveal that certain heterochromatic segments are associated with the nuclear membrane.--The nucleolar interstitial C-bands in A. sativum remain free in the nucleoplasm and may come close to each other due to heterochromatic attraction. Such a heterochromatic attraction is also evident between telomeric regions and between centromeres. However, a two by two attachment could not be noticed. A diagrammatic representation of the orientation of interphase chromosomes has been presented.     

Unusual chromosomal organization of telomeric sequences and expeditious karyotypic differentiation in the recently evolved Mus terricolor complex

The Mus terricolor complex displays a stable homozygous arrangement of autosomal heterochromatin variations in the form of accretion of definitive autosomal short arms among three nonoverlapping populations, in concert with an expeditious evolutionary differentiation into three chromosomal species: M. terricolor I, II, and III. In contrast to the highly conservative M. musculus-like chromosomes in the coexisting sibling species, M. booduga, reshuffling and differentiation of centric heterochromatin has occurred in harmony with a revision of centric configurations, resulting in acrocentric and submetacentric autosomes. The chromosomal distribution of the prevalent vertebrate telomeric sequence (TTAGGG)n was examined by fluorescence in situ hybridization to metaphase cells of M. terricolor I, II, and III. An unusual centric organization of internal telomeric sequences was detected in all the submetacentric and acrocentric autosomes. An auxiliary role of these presumably fragile, recombinogenic telomeric sequences in the evolutionary revision of centric configurations in the terricolor complex is hypothesized.     

Recapitulation of signals regulating embryonic bone formation during postnatal growth and in fracture repair

In this paper, we describe the results of a qualitative and quantitative study of chromosomal reorganizations observed in X-irradiated (1Gy and 2Gy) and cultured lymphocytes from Cebus apella. A total of 646 breakpoints have been detected, identified and localized in the ideogram of the species. The breakpoint distribution along chromosomes, p and q arms, and bands is not random. Chromosomes #11, #12 and chromosome arms 1p, 12p, 13p, 15p, 11q, and 12q are significantly more affected than expected, while chromosome #19 and chromosome arm 19q are less affected. Terminal regions of chromosome arms accumulate a higher number of breakpoints than the rest of the chromosome (37.82%). A high percentage (93.66%) of breakpoints is found in G negative bands.     

Chromosomes of Damaliscus (Artiodactyla, Bovidae): simple and complex centric fusion rearrangements

G- and C-banded karyotypes of Damaliscus hunteri, D. lunatus and D. pygargus were compared using the standard karyotype of Bos taurus. Chromosomal complements were 2n = 36 in D. lunatus jimela, 2n = 38 in D. pygargus phillipsi and D. p. pygargus, and 2n = 44 in D. hunteri. The fundamental number in all karyotypes was 60. Among the three species of Damaliscus, seven autosomal pairs and the X chromosomes were conserved. Y-chromosome differences were attributed to heterochromatic additions or deletions. Banded karyotypes of the two subspecies of D. pygargus exhibited complete homology. Chromosomal complements of D. pygargus and D. lunatus differed by a simple centric fusion. However, karyotypes of D. pygargus and D. lunatus differed from D. hunteri by numerous centric fusions, several of which were related by monobrachial chain complexes. Between the karyotypes of D. hunteri and D. pygargus or D. lunatus, there were two chain complexes, one involving five chromosomes (chain V) and the other involving 12 in pygargus (chain XII) or 13 in lunatus (chain XIII). There were also two simple centric fusions between D. hunteri and D. lunatus/D. pygargus; acrocentric chromosomes 13, 15, 20 and 22 in D hunteri were fused as 13;15 and 20;22 in D. lunatus and D. pygargus.     

Effects of cystamine on the state of peripheral blood neutrophilic granulocytes in vivo in rats

Chromosomal structural rearrangement in Paeonia brownii and P. californica (2n = 10) was studied by in situ hybridization using 18S rDNA probes. Six major rDNA sites were detected in mitotic cells of P. californica; six major and two minor rDNA sites were found in P. brownii. Two cytotypes (A and B), with different chromosomal morphology and (or) rDNA locations, were observed in the population of P. californica. Cytotype A, with rDNA sites only on the short arms of chromosomes, was considered to be the normal cytotype. Both translocation and pericentric inversion may have occurred to give rise to cytotype B, in which one homolog of chromosome 4 has rDNA sites on both arms while its homolog has no rDNA sites: one homolog of chromosome 3 has a rDNA site on the long arm. Two rearranged cytotypes, C and D, were observed in the population of P. brownii. Given that the normal cytotype of P. brownii is most likely to have six major rDNA sites on the short arms of chromosomes 3, 4, and 5, and two minor sites on the short arms of chromosome 2, cytotype C may have resulted from a translocation between the short arm of one homolog of chromosome 2 and the long arm of one homolog of chromosome 4, and cytotype D may have resulted from a translocation between the short arm of one homolog of chromosome 3 and the long arm of one homolog of chromosome 4. These results supported previous observations, based on meiotic configurations, that chromosomal structural rearrangement occurred frequently in P. brownii and P. californica.     

Activation PET scanning in pretreatment evaluation of patients with cerebral tumours or vascular lesions in or close to the sensorimotor cortex

Calmodulin is a ubiquitous transducer of calcium signals in eukaryotes. In diploid plant species, several isoforms of calmodulin have been described. Here, we report on the isolation and characterization of calmodulin cDNAs corresponding to 10 genes from hexaploid (bread) wheat (Triticum aestivum). These genes encode three distinct calmodulin isoforms; one isoform is novel in that it lacks a conserved calcium binding site. Based on their nucleotide sequences, the 10 cDNAs were classified into four subfamilies. Using subfamily-specific DNA probes, calmodulin genes were identified and the chromosomal location of each subfamily was determined by Southern analysis of selected aneuploid lines. The data suggest that hexaploid wheat possesses at least 13 calmodulin-related genes. Subfamilies 1 and 2 were both localized to the short arms of homoeologous-group 3 chromosomes; subfamily 2 is located on all three homoeologous short arms (3AS, 3BS and 3DS), whereas subfamily 1 is located only on 3AS and 3BS but not on 3DS. Further analysis revealed that Aegilops tauschii, the presumed diploid donor of the D-genome of hexaploid wheat, lacks a subfamily-1 calmodulin gene homologue, whereas diploid species related to the progenitors of the A and B genomes do contain such genes. Subfamily 3 was localized to the short arm of homoeologous chromosomes 2A, 2B and 2D, and subfamily 4 was mapped to the proximal regions of 4AS, 4BL and 4DL. These findings suggest that the calmodulin genes within each subfamily in hexaploid wheat represent homoeoallelic loci. Furthermore, they also suggest that calmodulin genes diversified into subfamilies before speciation of Triticum and Aegilops diploid species.     

Su(UR)ES: a gene suppressing DNA underreplication in intercalary and pericentric heterochromatin of Drosophila melanogaster polytene chromosomes

A genetic locus suppressing DNA underreplication in intercalary heterochromatin (IH) and pericentric heterochromatin (PH) of the polytene chromosomes of Drosophila melanogaster salivary glands, has been described. Found in the In(1)scV2 strain, the mutation, designated as Su(UR)ES, was located on chromosome 3L at position 34. 8 and cytologically mapped to region 68A3-B4. A cytological phenotype was observed in the salivary gland chromosomes of larvae homozygous and hemizygous for Su(UR)ES: (i) in the IH regions, that normally are incompletely polytenized and so they often break to form "weak points," underreplication is suppressed, breaks and ectopic contacts disappear; (ii) the degree of polytenization in PH grows higher. That is why the regions in chromosome arm basements, normally beta-heterochromatic, acquire a distinct banding pattern, i. e., become euchromatic by morphological criteria; (iii) an additional bulk of polytenized material arises between the arms of chromosome 3 to form a fragment with a typical banding pattern. Chromosome 2 PH reveals additional alpha-heterochromatin. Su(UR)ES does not affect the viability, fertility, or morphological characters of the imago, and has semidominant expression in the heterozygote and distinct maternal effect. The results obtained provide evidence that the processes leading to DNA underreplication in IH and PH are affected by the same genetic mechanism.     

A molecular cytogenetic analysis of X chromosome repatterning in the Bovidae: transpositions, inversions, and phylogenetic inference

Chromosomal homologies among the X chromosomes of species representative of eight bovid subfamilies and most of the recognized tribes were established using a combination of FISH and conventional G- and C-banding. Our analyses allowed for the delimitation of three X chromosome types represented, respectively, by cattle (Bovinae, tribe Bovini), the tragelaphines (Bovinae, tribe Tragelaphini), and a large assemblage comprising all the remaining subfamilies and their tribes (the Cephalophinae, Hippotraginae, Alcelaphinae, Antilopinae, Aepycerotinae, Peleinae, and Caprinae). The use of the bacterial artificial chromosome probe BAC 101 (which maps to Xp12 in cattle) and an Xp painting probe comprising sequences specific for the short arm of cattle Xp (Xp24-->p12) allowed us to orient this region, which has moved as a conserved euchromatic block during the evolution of the bovid X chromosome. We show that the differences between the three chromosomal types are attributable to a transposition, two inversions, and heterochromatic additions/deletions. A paucity of comparative mapping data precludes the assignment of the sequences contained in cattle Xp to either the presumed conserved (XCR) or the recently added (XAR) region of the eutherian X chromosome, and the reasons for the retention of these sequences as an evolutionarily conserved unit in the intrachromosomal restructuring of the bovid X across lineages remain enigmatic.     

The entire compound autosomes of Drosophila melanogaster

Three new unusual compound chromosomes have been synthesized in Drosophila melanogaster. They consist of two homologous autosomes joined together in the new order: right arm, left arm, centromere, left arm, right arm, for each of the two major autosomes, and one in which chromosomes 2 and 3 have been combined in the order: right arm of 2, left arm of 2, centromere, left arm of 3, right arm of 3. The attachments of the autosomal arms were accomplished by obtaining chromosome breaks at or very close to the ends of the left arms of the autosomes such that no essential chromosome material has been removed; the compounds derived from them are therefore referred to as entire compounds. These large chromosomes are recovered in progeny with frequencies lower than expectation partly because of zygote mortality associated with these chromosomes, and partly because of a failure of spermiogenesis.