The chromosomes of Afrotheria and their bearing on mammalian genome evolution.

Afrotheria is the clade of placental mammals that, together with Xenarthra, Euarchontoglires and Laurasiatheria, represents 1 of the 4 main recognized supraordinal eutherian clades. It reunites 6 orders of African origin: Proboscidea, Sirenia, Hyracoidea, Macroscelidea, Afrosoricida and Tubulidentata. The apparently unlikely relationship among such disparate morphological taxa and their possible basal position at the base of the eutherian phylogenetic tree led to a great deal of attention and research on the group. The use of biomolecular data was pivotal in Afrotheria studies, as they were the basis for the recognition of this clade. Although morphological evidence is still scarce, a plethora of molecular data firmly attests to the phylogenetic relationship among these mammals of African origin. Modern cytogenetic techniques also gave a significant contribution to the study of Afrotheria, revealing chromosome signatures for the group as a whole, as well as for some of its internal relationships. The associations of human chromosomes HSA1/19 and 5/21 were found to be chromosome signatures for the group and provided further support for Afrotheria. Additional chromosome synapomorphies were also identified linking elephants and manatees in Tethytheria (the associations HSA2/3, 3/13, 8/22, 18/19 and the lack of HSA4/8) and elephant shrews with the aardvark (HSA2/8, 3/20 and 10/17). Herein, we review the current knowledge on Afrotheria chromosomes and genome evolution. The already available data on the group suggests that further work on this apparently bizarre assemblage of mammals will provide important data to a better understanding on mammalian genome evolution.

Chromosome evolution in Xenarthra: new insights from an ancient group.

The Magnaorder Xenarthra is one of the four main supraordinal eutherian clades, together with Afrotheria, Euarchontoglires and Laurasiatheria. Xenarthra is an eminently Central and South American group of special interest in phylogenetic studies due to its possible position at the base of the eutherian tree. The use of modern cytogenetic techniques in some species of Xenarthra has provided important insights into the karyotypic evolution of mammals. Nevertheless, chromosome analyses in the group are still restricted, with only a few individuals of each species studied and karyotype descriptions mostly without banding patterns. In addition, it is likely that still unknown species exist and that the chromosome variability in the group is underestimated. We present a review of the currently available data on Xenarthra chromosomes and genomes and on the impact that their study has had in the understanding of mammalian genome evolution. It is clear that further cytogenetic analyses in Xenarthra, including banding patterns and molecular approaches, are likely to help in the identification of new species, reveal still undetected chromosome variations, provide information to support conservation strategies planning, and greatly contribute to a better understanding of mammalian genome evolution.

Chromosome painting in Callicebus lugens, the species with the lowest diploid number (2n=16) known in primates.

Cytogenetic studies have shown that New World primates are karyologically diverse and highly derived. The genus Callicebus is the best example of this karyological diversity, with diploid numbers ranging from 2n=50 to 2n=16. We report on Callicebus lugens, which has the lowest diploid number (2n=16) yet found in the primate order and represents a striking example of extreme karyotypic shuffling. To better understand the genomic rearrangements that have resulted in this extremely low diploid number, we mapped chromosome homologies between C. lugens and humans by in situ hybridization. The total number of hybridization signals was 42, excluding the Y chromosome, with a total of 34 syntenic associations not found in humans. This species has one of the most derived karyotypes among the Platyrrhini. Fusion has been the predominant mode of karyological evolution, although fissions and inversions have also transformed the C. lugens karyotype. Remarkably in such a highly rearranged karyotype, the synteny of 11 human chromosomes (4, 5, 9, 12, 13, 14, 17, 18, 20, 21, and X) was maintained intact, even if most of these human-homologous gene clusters were translocated. Other human syntenies, such as homologues to human chromosomes 10 and 16, were highly fragmented. Comparisons of the C. lugens-human homology map with those of other New World primates have not yet helped establish a phylogenic arrangement between congeneric species or link Callicebus with any other genus.

Cloning and mapping of platypus SOX2 and SOX14: insights into SOX group B evolution.

Group B SOX genes, the closest relatives to the sex-determining gene SRY, are thought to have evolved from a single ancestral SOX B by a series of duplications and translocations. The two SOX B genes SOX2 and SOX14 co-localize to chromosome 3q in humans. SOX2 and SOX14 homologues were cloned and characterized in the platypus, a monotreme mammal distantly related to man. The two genes were found to co-localize to chromosome 1q in this species. Proximity of the two related genes has therefore been conserved for 170 Myr, since humans and platypus diverged. The sequence similarity and conserved synteny of these group B genes provide clues to their origin. A simple model of SOX group B gene evolution is proposed.

Karyotype relationships between distantly related marsupials from South America and Australia.

Reciprocal chromosome painting and G-banding were used to compare the karyotypes of three Australian marsupials (Sminthopsis crassicaudata, Macropus eugenii, Trichosurus vulpecula) and one South American marsupial (Monodelphis domestica). The results revealed only a limited number of rearrangements between these species and that the four karyotypes can be described as different combinations of fifteen conserved segments. Five chromosomes are totally conserved between M. domestica (pairs 1, 2, 5, 8 and the X) and the presumed 2n = 14 Australian ancestral karyotype, while M. domestica pairs 3 and 6 and 4 and 7 would have been involved in fusion/fission rearrangements. Chromosome comparisons are presented in a chromosome homology map. Although the species studied diverged 70 million years ago, the karyotype of Monodelphis domestica is highly conserved in relation to those of Australian marsupials.

Deletion of the factor IX gene as a result of translocation t(X;1) in a girl affected by haemophilia B.

A balanced de novo translocation t(X;1) is described in a girl with severe haemophilia B. The translocated X was shown cytologically to be preferentially active, and methylation analysis of the DXS255 locus confirmed the skewed X-inactivation with the paternal allele being the active one. Cytogenetic and molecular analysis showed that this chromosomal rearrangement led to the deletion of at least part of the factor IX gene. Therefore, the girl was heterozygous for factor IX deficiency and expression of her clinical phenotype was the result of the inactivation of the normal maternal X chromosome. The localization of one of the X chromosome translocation breakpoints in YAC clone 957F9, that was demonstrated to map distally to the factor IX gene, revealed the complexity of this chromosomal rearrangement.

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.

Comparative genome analysis in American marsupials: chromosome banding and in-situ hybridization.

We performed a comparative analysis of the G- and C-banded karyotypes of seven species of didelphid marsupials, representing the three diploid numbers (2n = 14, 18 and 22) known to occur in this family. In addition to a great similarity among karyotypes with the same diploid numbers, we also identified homeologies for all autosomal arms comprising the three karyotypes. Robertsonian rearrangements, pericentric inversions and heterochromatin variation account for the differences among the karyotypes. Interspecific variation in the size of the sex chromosomes is due to differences in heterochromatic content. In-situ hybridization with total genomic DNA revealed considerable conservation of the euchromatic portions of the three karyotypes and indicated divergence of repetitive DNA sequences in autosomal heterochromatin.

Karyotype evolution of marsupials: from higher to lower diploid numbers.

A basic 2n = 14 ancestral marsupial karyotype giving rise to higher diploid numbers through chromosome fissions has been widely accepted for the last three decades. Our finding of interstitial telomeres in two South American species, one with the 2n = 14 "ancestral karyotype" and the other with 2n = 18, indicates that these complements evolved from a karyotype with a higher diploid number. A new scenario for the karyotype evolution in the group is put forward. In this scenario an ancestral karyotype with at least 22 chromosomes would have originated the basic karyotype with 2n = 14 before the radiation of marsupials.

Clinical healing two to six years after poststreptococcal glomerulonephritis in Trinidad.

To determine the incidence of chronic nephritis after poststreptococcal acute glomerulonephritis in Trinidad, 760 patients (41 adult) were examined two to six years after recovery from the illness, 344 being studied twice (four and six years). Only 1.8 per cent had persistent urine abnormalities on their last follow-up examination, and another 8.0 per cent had abnormalities that were transient or occurred only after the patient had assumed the lordotic position. In 1.4 per cent hypertension was present, whereas only one had azotemia. Both persistent urine abnormalities and hypertension increased in prevalence with age at onset of prior poststreptococcal glomerulonephritis but did not vary between sexes, races or epidemic versus endemic forms. Half the urine abnormalities present four years after recovery were absent two years later. Thus, poststreptococcal acute glomerulonephritis appears to have a low incidence of chronicity in Trinidad, with continuing resolution for more than four years.

Tropical acute rheumatic fever and associated streptococcal infections compared with concurrent acute glomerulonephritis.

Ninety-three patients with acute rheumatic fever and 195 patients with acute glomerulonephritis were observed in Trinidad during an outbreak of scabies with a high incidence of secondary streptococcal infections. Clinical and laboratory manifestations of ARF were the same as those seen in temperate zones, except that antistreptolysin O titers were less markedly increased. The patients with ARF were similar to those with AGN in respect to sex, race, location of residence, and living conditions, but were older and had markedly fewer skin infections. Currently prevalent nephritogenic streptococcal strains never were isolated from patients with ARF even when M55 streptococci appeared and led to an epidemic of AGN.

The families of patients with acute rheumatic fever or glomerulonephritis in Trinidad.

The families of 21 patients with acute rheumatic fever (ARF) and 44 patients with acute glomerulonephritis (AGN) in Trinidad were examined in their homes. The ARF and AGN families were equally large and crowded and they lived in the same largely rural areas. However, only 22% of the ARF family members had skin infections in contrast to 61% of the AGN family members. Sixty-eight per cent of skin infections in ARF families and 69% of skin infections in Agn families yielded group A streptococci. Throat cultures were positive in 19% of ARF family members and 25% of AGN family members. Thirty-two per cent of 51 group A strains isolated from ARF family members (29 from throat, 22 from skin) were M11 or "M41" strains which were associated with ARF during the study, while only 8% were M1, T4 (MNT or 60) or M55 strains which were associated with AGN. In contrast, 49% of 171 group A strains isolated from Agn family members (64 from throat, 107 from skin) were M1, T4 (MNT or 60) or M55 while only 10% were M11 or "M41." Serum antibody titers were similar in both groups: antistreptolysin-0 titers were not markedly increased in either while anti-hyaluronidase and/or antideoxyribonuclease-B titers were increased in both. Evidence of subclinical AGN was found equally often in both groups: 6% of each had abnormal urine and 4% of each had decreased serum complement while 2% of the ARF and 3% of the AGN family members had both abnormal urine and decreased serum complement.

Immunoglobulins and complement components in synovial fluid of patients with acute rheumatic fever.

Three components of complement and six other serum proteins were assayed in synovial fluid and serum samples from 25 patients with acute rheumatic fever in Trinidad. The resulting data indicate a relative decrease in both early and late components of complement within the synovial fluids which suggests local activation by immune complexes. Such activation of complement within the joint spaces may play a primary role in development of the inflammatory arthritis of acute rheumatic fever.

Salivary immunoglobulins and streptococcal antibodies in patients with acute rheumatic fever.

Evidence of a local immunologic response to recent pharyngeal infection with group A streptococci was sought in 23 patients with acute rheumatic fever. Salivas from these patients were examined and compared with salivas from control subjects. As assayed, both IgG and IgA values ranged between 8 and 170 mg. per 100 ml. in 10-fold concentrated samples of saliva. When these values were corrected for the concentration of the samples and for the use of serum IgA standards, the following mean values were obtained: for patients with acute rheumatic fever, IgG was 4.5 mg. per 100 ml. and IgA was 8.7 mg. per 100 ml.; for control subjects, IgG was 5.0 mg per 100 ml. and IgA was 9.9 mg. per 100 ml. IgM was present in amounts of 1 to 2 mg. per 100 ml. in 2 patients and 7 control subjects. Antistreptolysin O titers correlated with amounts of IgG in the saliva samples while antihyaluronidase, antigroup A carbohydrate, and antiteichoic acid titers did not. None of the antibody titers correlated with the IgA content of the saliva samples. Furthermore, saliva antibody titers did not differ between patients with acute rheumatic fever and control subjects.