Bone health and SATB2-associated syndrome.

SATB2-associated syndrome (SAS) is a rare disorder caused by alterations in the special AT-rich sequence-binding protein 2 (SATB2). Skeletal abnormalities such as tibial bowing, osteomalacia, osteopenia or osteoporosis have been reported suggesting a higher frequency of skeletal complications in SAS. The optimal timing, necessity, and methodology for routine assessment of bone health in individuals with SAS, however, remain unclear. We report molecular and phenotypic features of 7 individuals with SAS documented to have low bone mineral density (BMD) ascertained by dual-energy X-ray absorptiometry (DXA), often preceded by tibial bowing. The lowest BMD Z-scores ranged -2.3 to -5.6. In 4 individuals, total alkaline phosphatase levels were elevated (2 with elevated bone fraction) around the time of low BMD documentation. A clinically significant fracture history and a diagnosis of pediatric osteoporosis were present in 4 individuals. Pamidronate treatment in 2 children improved BMD. In conclusion, low BMD, fractures, and tibial bowing are relatively common skeletal complications in individuals with SAS. DXA is a useful tool when evaluating a child with SAS suspected to have low BMD and the results might alter clinical management.

Rapid identification of overlapping YACs in the MEN2 region of human chromosome 10 by hybridization with Alu element-mediated PCR products.

An overlapping set of 21 yeast artificial chromosomes (YACs) spanning the RET proto-oncogene [Takahashi et al., Oncogene 3 (1988) 571-578] and D10S102 markers on human chromosome 10 was isolated in a series of hybridization-based chromosomal walks in a YAC library. Genetic linkage analyses implicate this chromosomal region as the location of the gene (MEN2A) responsible for multiple endocrine neoplasia type 2A. Four YACs carrying a RET sequence-tagged site (STS) and two YACs carrying a D10S102 STS were used to initiate chromosome walks. These were based on hybridization of Alu element-mediated polymerase chain reaction (Alu-PCR) products from YACs to dot blots of Alu-PCR products from complex pools of YAC clones. The hybridization anchor content of YACs identified in the walks was confirmed by probing blots of Alu-PCR products from individual YACs and by comparing Alu-PCR fingerprints of each YAC. Ten hybridization-based Alu-PCR anchors and three STS anchors were ordered within eleven intervals created by the 21 overlapping YACs. The order of anchors requiring the fewest gaps in the YACs is consistent with the walking results and establishes the STS anchor order as D10S102-D10S94-RET. The overlapping set of YACs represents about 1.55 Mb of the human genome according to restriction mapping of four representative YACs in the contig. These results demonstrate the power of Alu-PCR hybridization for chromosomal walking and provide a rich source of overlapping YACs which can be used to identify candidate MEN2A genes.

Infection of Peruvian Aotus nancymai monkeys with different strains of Plasmodium falciparum, P. vivax, and P. malariae.

Aotus nancymai (karyotype I) monkeys from Peru were studied for their susceptibility to infection with Plasmodium falciparum, P. vivax, and P. malariae. Three strains of P. falciparum (Santa Lucia from El Salvador, Indochina I/CDC from Thailand, and Uganda Palo Alto) were inoculated into 38 monkeys. The results indicated that this species of Aotus monkey is highly susceptible to infection. The Uganda Palo Alto and the Santa Lucia strain parasites appear to be the most useful for immunologic and chemotherapeutic studies. Five strains of P. vivax (Chesson, ONG, Vietnam Palo Alto, Salvador I, and Honduran I/CDC) were inoculated into 28 monkeys. The Vietnam Palo Alto strain produced the highest level parasitemias ranging from 23,800 to 157,000/mm3. Mosquito infections were obtained with the ONG, Chesson, and Salvador I strains. Two out of 6 attempts to transmit P. vivax via sporozoite inoculation to splenectomized monkeys were successful with prepatent periods of 39 and 57 days. Five monkeys were infected with the Uganda I/CDC strain of P. malariae. Maximum parasitemias ranged from 10 to 5,390/mm3.

Infection of Aotus vociferans (karyotype V) monkeys with different strains of Plasmodium vivax.

Twenty splenectomized Aotus vociferans (karyotype V) monkeys were infected with strains of Plasmodium vivax from New Guinea, North Korea, Indonesia, El Salvador, and Honduras. Peak parasite densities ranged from 4,840 to 75,500 per mm3. Gametocytes infective to different species of mosquitoes were produced with all strains of P. vivax studied. Two transmissions of the Chesson strain of P. vivax were made by the intravenous inoculation of dissected sporozoites from An. dirus mosquitoes. Prepatent periods were 16 days.

Infection of Aotus vociferans monkeys with different strains of Plasmodium falciparum.

Twenty-one splenectomized Aotus vociferans monkeys were infected with the different strains/clones of Plasmodium falciparum. Maximum parasitemia ranged from 1,302 to 1,460,000 parasites per mm3. Only the Santa Lucia strain was shown to produce gametocytes for extended periods. Gametocytes produced during the primary episode of parasitemia were highly infective to Anopheles freeborni mosquitoes. Gametocytes produced during recrudescence were not infective to mosquitoes feeding directly on the animals. This lack of mosquito infection during recrudescence periods suggests the presence of transmission-blocking immunity, which may be important in understanding the control of malaria through immunologic initiatives.

Studies on the North Korean strain of Plasmodium vivax in Aotus monkeys and different anophelines.

Twenty-two Aotus monkeys of different karyotypes were infected with the North Korean strain of Plasmodium vivax. Aotus lemurinus griseimembra animals from Colombia produced higher maximum parasitemias and more readily infected mosquitoes than did Aotus monkeys from Bolivia (K-VI) or Peru (K-V and K-X). Comparative feedings indicated that the most susceptible mosquito species was Anopheles stephensi, followed by An. gambiae, An. dirus, An. freeborni, An. quadrimaculatus, An. culicifacies, and An. maculatus.

Confirmed assignments of 15 structural gene loci to chromosomes of four owl monkey karyotypes.

Fifteen gene loci for constitutive enzymes previously localized to specific owl monkey chromosomes of karyotypes III, V, and VI are confirmed by their assignments to homologous chromosomes of owl monkey karyotypes I, II, IV, and VII. The syntenic mapping of LDHA and GPI on a large metacentric, II-2, and the separate assignment of these two loci to two acrocentrics, I-9 and I-15, provide genetic evidence supporting the proposed fusion-fission event that characterized the karyotypic difference between owl monkeys inhabiting Colombia and the Panama Canal Zone. Moreover, the proposed hypothesis on chromosome polymorphism among the Colombian owl monkeys with karyotypes II, III, and IV, resulting from a fusion-fission event involving one metacentric and two subtelocentric pairs, is supported by the assignment of LDHB and MDH1 to the large metacentric I-2 and the separate localization of these two gene loci to II-13 and II-14, respectively.

[Changes in malondialdehyde contents in serum and tissues after ischemia and reperfusion of the bowel in dogs].

The portal circulation was reduced to 50% for one hour by partially occluding the superior mesenteric artery, and the relationship between the changes in malondialdehyde (MDA) and multiple organ damage was studied. The results showed that there was a marked change in hepatic enzymology, with MDA values significantly elevated in serum and lung and liver tissues after the reperfusion when compared with controls. These results suggested that lipid peroxidation initiated by free oxygen radicals played an important role in the multiple organ damages, and the extent of organ damage was closely correlated with the contents of MDA in tissues.

Regional localization of the owl monkey MYB and OMLA gene loci.

Regional mapping of the MYB and OMLA gene loci to their respective sites on owl monkey chromosome 9q of karyotype VI support the proposed homology of this primate chromosome with the human chromosome 6 and the premise that structural rearrangement on this putative homolog occurred during evolutionary divergence of the higher primates.

Owl monkey gene loci for c-fes and albumin are syntenic.

Segregation analysis of rodent-owl monkey somatic cell hybrids probed with a human c-fes DNA sequence disclosed the owl monkey c-fes locus to be syntenic with the albumin loci on chromosome 1 of karyotype VI, chromosome 12 of karyotype V, and chromosome 9 of karyotype IV. This finding substantiates the proposed homology of these chromosomes and demonstrates the complexity of rearrangement of individual genes during chromosomal phylogeny which is not detectable by morphological comparison alone.

Synteny of SIS and IGLC in owl monkeys. Evidence for karyotype evolution.

Two cloned human DNA sequences specific for the SIS and IGLC genes were hybridized with DNAs from panels of rodent-owl monkey somatic cell hybrids. Independent segregation analysis of the related owl monkey DNA sequences for these two genes demonstrated that both the SIS locus and the IGLC locus are localized on owl monkey chromosome 3 of karyotype VI, chromosome 3 of karyotype V, and chromosome 6 of karyotype IV. These syntenic assignments provide genetic evidence for the homology of, at least, a portion of these chromosomes from three owl monkey karyotypes. The findings also help to redetermine the possible chromosome rearrangement that characterized the chromosome complement of owl monkey from Panama and northwestern Colombia with karyotypes II, III, IV, VIII, and IX from other owl monkey populations with karyotypes I, V, VI, VII, X, and XI.

Comparative gene mapping of the owl monkey, Aotus, with karyotypes V (2n = 46) and VI (2n = 49, 50).

Biochemical analysis of cytogenetically defined rodent--owl monkey hybrid cell lines segregating owl monkey chromosomes allowed the assignment of 14 structural gene markers to seven owl monkey karyotype V (K-V, 2n = 46) chromosomes. Comparative study of assigned gene loci to chromosomes of owl monkey karyotype V (K-V, 2n = 46) and karyotype VI (K-VI, 2n = 49/50) provides the first evidence supporting some of the previously proposed mechanics of chromosomal rearrangements responsible for the great karyotypic diversity in owl monkeys. In addition, chromosome homologies deduced from gene loci of Aotus and other primates are compared here to determine the conservation or modification of chromosome morphology in the course of primate evolution. The data demonstrate the usefulness of gene mapping as an alternative to banding patterns for obtaining evidence on chromosome evolution.

Gene map of the new world Bolivian owl monkey, Aotus.

Somatic cell hybrids between rodent cells and owl monkey cells preferentially segregating primate chromosomes were selected for mapping owl monkey genes. Concordant segregation of phenotype and chromosome in 35 independent hybrid clones and 105 subclones with different reduced owl monkey complements provided evidence for the assignment of 17 gene loci to 9 chromosomes of the Bolivian owl monkey with a karyotype VI (2n = 49 male, 50 female). The assignments include: MDH1 to chromosome 2, MDH2 to chromosome 4, FH to chromosome 6, LDHB to chromosome 10, NP-MPI-PKM2-SORD to chromosome 11, PGM1-PGD-AK2-ENO1 to chromosome 12, LDHA to chromosome 19, GPI to chromosome 25 and PGK-G6PD-GLA to the X chromosome.

A putative homoeolog of human chromosome 12 in the owl monkey.

Analyses of Southern blots of rodent x owl monkey somatic cell hybrids permitted syntenic assignment of gene loci coding for triosephosphate isomerase (TPI), antigen CD4(T4), Kirsten rat sarcoma 2(KRAS2) virus, insulin-like growth factor 1 (IGF1), and alpha 2-macroglobulin (A2M) to chromosome 10 of owl monkey karyotype VI(2n = 49, 50). In addition, regional in situ localization of the T4 and KRAS2 loci on the proximal region of the long arm of this acrocentric chromosome and on the corresponding homologous region on the long arm of metacentric chromosome 1 of karyotype IV (2n = 52) substantiated our hypothesis that a single fusion or fission event is responsible for the polymorphism in chromosome number characteristic of owl monkeys from at least three allopatric populations. The study supports a putative homoeology between owl monkey chromosome 10 (K-VI) and human chromosome 12. The morphological differences between these two primate chromosomes indicate evolutionary rearrangements involving at least one pericentric inversion.

Chromosome assignment of the owl monkey MOS and MYC gene loci.

Southern blot hybridizations of rodent x owl monkey hybrid DNAs with human cDNA probes allowed the mapping of the MOS and MYC gene loci to owl monkey chromosome 16 of karyotype VI (2n = 49 male/50 female) and to the homologous chromosome 15 of karyotype V (2n = 46). Synteny of MOS and MYC gene loci in both man and owl monkey suggests this chromosome segment's conservation in primates, contrasting with its disruption in the mouse.

Comparative chromosomal mapping of the owl monkey serum albumin gene.

We have mapped the albumin locus (ALB) in the owl monkey, Aotus trivirgatus, using a cloned human albumin gene probe, pcHSA 33-1. Rodent-owl monkey somatic cell hybrids were used to map the owl monkey albumin locus in three subgroups of Aotus, karyotypes II, V, and VI. Segregation analysis of the molecular hybridization pattern of pcHSA 33-1 in the somatic cell hybrids indicated that the albumin locus maps to chromosome 9 of owl monkey karyotype II, chromosome 12 of karyotype V, and chromosome 1 of karyotype VI. This assignment provides evidence for the homology of these three chromosomes and supports the hypothesis of Ma on the formation of chromosome 1 in karyotype VI.

Chromosome mapping of the owl monkey CSF1R and IL5 genes.

We mapped the owl monkey colony-stimulating factor 1 receptor (CSF1R) locus to the proximal region of chromosome 3q of karyotype VI(K-VI) and karyotype V(K-V) and the interleukin 5 (IL5) locus to the mid-region of chromosome 3q(K-VI) and 19q(K-IV) using a combination of Southern hybridization of somatic cells and in situ chromosomal hybridization methodologies. The findings support the proposed evolution of owl monkey chromosome 3(K-VI) from a fusion of two smaller structures, the homologs of chromosomes 6 and 19 (K-IV). The data also indicate genomic conservation of the HSA 5q23-q35 segment in the higher primates.

Mapping of five human chromosome 19 DNA markers to owl monkey chromosomes.

Hybridization of DNA from three panels of karyotypically distinct owl monkey x rodent somatic cell hybrids with human DNA probes resulted in the syntenic assignments of INSR-LDLR-TGFB1-APOE-D19S8 to owl monkey chromosome 25 of karyotype VI (2n = 49/50), INSR-LDLR-TGFB1-D19S8 to chromosome 2 of karyotype II (2n = 54), and INSR-APOE to chromosome 2 of karyotype V (2n = 46). The APOE and D19S8 loci are on adjacent regions proximal to the centromere of chromosomes 25q (K-VI) and 2p (K-II), as determined by in situ chromosomal hybridization analysis. These findings support our previous proposals on (1) the homology of these chromosomes of three owl monkey karyotypes, (2) the evolutionary derivation of chromosome 2 of karyotypes II and V as the result of two separate centric fusion events, and (3) the likelihood that owl monkey chromosome 25 (K-VI) (and its homologs) is a conserved genetic homoeolog of human chromosome 19.

Added heterochromatin segments in chromosomes of squirrel monkeys (Saimiri sciureus).

Variant structural homozygosity, observed previously only in plants and insects, is reported herein for the first time in mammalian chromosomes. The addition of a heterochromatic chromosome segment to either one or both homologues is described in autosomes B5 and B12 of the squirrel monkey, Saimiri sciureus.