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The phylogeny of the hominoid primates, as indicated by DNA-DNA hybridization.

The living hominoid primates are Man, the chimpanzees, the Gorilla, the Orangutan, and the gibbons. The cercopithecoids (Old World monkeys) are the sister group of the hominoids. The composition of the Hominoidea is not in dispute, but a consensus has not yet been reached concerning the phylogenetic branching pattern and the dating of divergence nodes. We have compared the single-copy nuclear DNA sequences of the hominoid genera using DNA-DNA hybridization to produce a complete matrix of delta T50H values. The data show that the branching sequence of the lineages, from oldest to most recent, was: Old World monkeys, gibbons, Orangutan, Gorilla, chimpanzees, and Man. The calibration of the delta T50H scale in absolute time needs further refinement, but the ranges of our estimates of the datings of the divergence nodes are: Cercopithecoidea, 27-33 million years ago (MYA); gibbons, 18-22 MYA; Orangutan, 13-16 MYA; Gorilla, 8-10 MYA; and chimpanzees-Man, 6.3-7.7 MYA.

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Stomach lysozyme gene of the langur monkey: tests for convergence and positive selection.

Genomic blotting and enzymatic amplification show that the genome of the langur monkey (like that of other primates) contains only a single gene for lysozyme c, in contrast to another group of foregut fermenters, the ruminants, which have a multigene family encoding this protein. Therefore, the langur stomach lysozyme gene has probably evolved recently (i.e., within the period of monkey evolution) from a conventional primate lysozyme. The sequences of cDNAs for the stomach lysozyme of langur and the conventional lysozymes of three other Old World monkeys were determined. Identification of the promoter for the stomach gene and comparison to the human gene, which is expressed conventionally in macrophages, show that both lysozyme genes use the same promoter. This suggests that the difference in expression patterns is due to change(s) in enhancer or silencer regulatory elements. With the cDNA sequences the hypothesis that the langur stomach lysozyme has converged in amino acid sequence upon the stomach lysozymes of ruminants is tested. Consistent with the convergence hypothesis, only those sites that specify amino acids in the mature lysozyme are shared uniquely with ruminant lysozyme genes. None of the silent sites at third positions of codons or in noncoding regions support a link between the langur and ruminants. Statistical analysis based on silent sites rules out the possibility of horizontal transfer of a stomach lysozyme gene between the langur and ruminant lineages and supports the close relationship of the langur lysozyme gene to that of other monkeys.

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DNA hybridization evidence of hominoid phylogeny: results from an expanded data set.

The living hominoids are human, the two species of chimpanzees, gorilla, orangutan, and nine species of gibbons. The cercopithecoids (Old World monkeys) are the sister group of the hominoids. A consensus about the phylogeny of the hominoids has been reached for the branching order of the gibbons (earliest) and the orangutan (next earliest), but the branching order among gorilla, chimpanzees, and human remains in contention. In 1984 we presented DNA-DNA hybridization data, based on 183 DNA hybrids, that we interpreted as evidence that the branching order, from oldest to most recent, was gibbons, orangutan, gorilla, chimpanzees, and human. In the present paper we report on an expanded data set totaling 514 DNA hybrids, which supports the branching order given above. The ranges for the datings of divergence nodes are Old World monkeys, 25-34 million years (Myr) ago; gibbons, 16.4-23 Myr ago; orangutan, 12.2-17 Myr ago; gorilla, 7.7-11 Myr ago; chimpanzees-human, 5.5-7.7 Myr ago. The possible effects of differences in age at first breeding are discussed, and some speculations about average genomic rates of evolution are presented.

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Analysis of the organisation and localisation of the FSHD-associated tandem array in primates: implications for the origin and evolution of the 3.3 kb repeat family.

The D4Z4 locus is a polymorphic tandem repeat sequence on human chromosome 4q35. This locus is implicated in the neuromuscular disorder facioscapulohumeral muscular dystrophy (FSHD). The majority of sporadic cases of FSHD are associated with de novo DNA deletions within D4Z4. However, it is still not known how this rearrangement causes FSHD. Although the repeat contains homeobox sequences, despite exhaustive searching, no transcript from this locus has been identified. Therefore, it has been proposed that the deletion may invoke a position effect on a nearby gene. In order to try to understand the role of the D4Z4 repeat in this disease, we decided to investigate its conservation in other species. In this study, the long-range organisation and localisation of loci homologous to D4Z4 were investigated in primates using Southern blot analysis, pulsed field gel electrophoresis and fluorescence in situ hybridisation. In humans, probes to D4Z4 identify, in addition to the 4q35 locus, a closely related tandem repeat at 10qter and many related repeat loci mapping to the acrocentric chromosomes; a similar pattern was seen in all the great apes. In Old World monkeys, however, only one locus was detected in addition to that on the homologue of human chromosome 4, suggesting that the D4Z4 locus may have originated directly from the progenitor locus. The finding that tandem arrays closely related to D4Z4 have been maintained at loci homologous to human chromosome 4q35-qter in apes and Old World monkeys suggests a functionally important role for these sequences.

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Lipoprotein(a): nonhuman primate models.

Lipoprotein(a) [Lp(a)] is a low density lipoprotein which has apo(a) disulfide-linked to apoB100. Apo(a) has recently been shown to have a striking homology with plasminogen, a knowledge that has stimulated a lot of interest in the mechanism of atherogenicity and thrombogenicity of this lipoprotein particle. Several studies have documented the presence of Lp(a) in nonhuman primates with particular reference to the rhesus monkeys and baboons. The Lp(a) of rhesus monkey is structurally very similar to that of humans, except for the absence of kringle V and the amino acid composition of the catalytic region. The Lp(a) of nonhuman primates, like their human counterparts, exhibit a wide range of interindividual plasma levels and also a wide size polymorphism of apo(a). Nonhuman primates appear to represent a good model for the study of the structure and biology of Lp(a).

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Measuring fruit patch size for three sympatric indonesian primate species.

Food availability is one of the basic factors affecting primate density and socioecology, but food availability is difficult to assess. Two different ways to obtain accurate estimates of food availability have been proposed: using phenology data or using the behaviour of animals. Phenology data can be refined by only including trees that are large enough to be used; including (potential) tree species in which by the concerned primate species forage; or including (fruiting) trees of these species that actually produce fruit. Alternatively, the sizes of the actually visited trees (foraging trees) give an estimate of fruit availability. These measures are compared for three sympatric primate species at the Ketambe Research Station, Sumatra, Indonesia: the Thomas langur, the long-tailed macaque and the orangutan. The sizes of fruiting trees and the foraging trees are larger than the potential trees. The sizes of the potential trees and of the fruiting trees are similar for the three primate species. This, however, is not reflected in the use of trees: the langurs forage on average in trees of similar size to those producing fruit, whereas the macaques and orangutans forage in trees larger than those producing fruit. The use of trees does not necessitate a different cut off point of included dbhs for the three compared primate species. The use of trees of different sizes, however, may be regulated by food competition. This indicates that sympatric primates make different foraging decisions and that behavioural measures of food availability will be less reliable.

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The seasonal daily travel in a group of Sichuan snub-nosed monkey (Pygathrix roxellana) in Shennongjia Nature Reserve, China.

The seasonal daily travel of a group of snub-nosed monkey (Pygathrix roxellana) was investigated using the group's straight-line distance method. The group was followed from dawn to dusk for 30 consecutive days during each season to encompass all aspects of daily travel patterns. The results showed that in Summer and Autumn, the mean daily travel distance (m) was significantly longer than in Winter and Spring. There was no significant difference in the mean daily travel distance between Summer and Autumn or between Winter and Autumn. The mean travel distance (m/hr) during daytime was significantly higher in Summer and Autumn than in Spring, and in Autumn than in Winter. The travel of the group in all seasons had similar patterns. Traveling showed morning and afternoon peaks, with a rest period at the noon in a day. Seasonal food availability, length of daytime, and mean travel distance (m/hr) during daytime might have contributed to the different daily travel distances in different seasons.

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Distribution, population density, and status of sympatric cercopithecids in the Campo-Ma'an area, Southwestern Cameroon.

A study on species composition, distribution, and population density of cercopithecids in the Campo-Ma'an area, Southwestern Cameroon, was undertaken from December 1997 until August 2000. A total of 665.5 km of line transects was used for the census. Thirteen diurnal primate species including five endangered species (Gorilla g. gorilla, Pan troglodytes, Mandrillus sphinx, Colobus satanas, Cercocebus torquatus) were recorded in the Campo Forest, the greatest part of which is a logging concession. Cercopithecus nictitans (1.43 groups/km2), C. cephus (1.13 groups/km2), C. pogonias (0.81 groups/km2), and C. torquatus (0.51 groups/km2) occurred at medium frequencies compared to figures from other Central African study sites. Mandrill densities estimated (0.27 groups/km2) show that the area is very important for the conservation of this rare species. Guenon densities found inside areas with a high level of human activities did not differ significantly from densities estimated in areas with a lower level of human activities. C. torquatus densities were significantly higher in the areas with a low level of human disturbance and encounter rates with Lophocebus albigena also indicate a preference of less disturbed areas. Mangabeys are thus likely to be adversely affected by human activities in the area. In the Ma'an Forest, which has not been logged yet, ten species were confirmed. The population densities of two guenons (C. nictitans and C. cephus) were estimated to be twice as high in the unlogged forest area as compared to the logged forest of Campo. Other species are rarer in the Ma'an Forest than in the Campo Forest. Although mangabeys are adversely affected by human activities, the results still indicate that selective logging may be compatible with the conservation of cercopithecids, if a reduced damage logging concept and antipoaching measures are implemented. Increased hunting activities following logging operations will definitely have a negative longterm impact on primate populations in the Campo-Ma'an area if no further, more effective conservation measures will regulate wildlife use in future.

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Autoradiographic study of 14C-sulpiride in monkey.

Substituted benzamides have been the object of numerous metabolic studies including many by whole body autoradiography of rats and mice. The present study reports autoradiographic data concerning 14C-labelled Sulpiride in monkey. The Study was limited to the brain in order to elucidate the controversial question as to whether the drug can cross the blood-brain barrier. The results showed that in monkey, as in rat and mouse, there is no localization in the brain as can be clearly seen on the autoradiograms. In view of these results and of the undeniable neuroleptic properties of Sulpiride, an indirect mode of action through the release of endogenous mediators is proposed.

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Molecular timing of primate divergences as estimated by two nonprimate calibration points.

The complete mitochondrial DNA (mtDNA) molecule of the hamadryas baboon, Papio hamadryas, was sequenced and included in a molecular analysis of 24 complete mammalian mtDNAs. The particular aim of the study was to time the divergence between Cercopithecoidea and Hominoidea. That divergence, set at 30 million years before present (MYBP) was a fundamental reference for the original proposal of recent hominoid divergences, according to which the split among gorilla, chimpanzee, and Homo took place 5 MYBP. In the present study the validity of the postulated 30 MYBP dating of the Cercopithecoidea/Hominoidea divergence was examined by applying two independent nonprimate molecular references, the divergence between artiodactyls and cetaceans set at 60 MYBP and that between Equidae and Rhinocerotidae set at 50 MYBP. After calibration for differences in evolutionary rates, application of the two references suggested that the Cercopithecoidea/Hominoidea divergence took place >50 MYBP. Consistent with the marked shift in the dating of the Cercopithecoidea/Hominoidea split, all hominoid divergences receive a much earlier dating. Thus the estimated date of the divergence between Pan (chimpanzee) and Homo is 10-13 MYBP and that between Gorilla and the Pan/Homo linage approximately 17 MYBP. The same datings were obtained in an analysis of clocklike evolving genes. The findings show that recalculation is necessary of all molecular datings based directly or indirectly on a Cercopithecoidea/Hominoidea split 30 MYBP.

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Ancestral population sizes and species divergence times in the primate lineage on the basis of intron and BAC end sequences.

The effective sizes of ancestral populations and species divergence times of six primate species (humans, chimpanzees, gorillas, orangutans, and representatives of Old World monkeys and New World monkeys) are estimated by applying the two-species maximum likelihood (ML) method to intron sequences of 20 different loci. Examination of rate heterogeneity of nucleotide substitutions and intragenic recombination identifies five outrageous loci (ODC1, GHR, HBE, INS, and HBG). The estimated ancestral polymorphism ranges from 0.21 to 0.96% at major divergences in primate evolution. One exceptionally low polymorphism occurs when African and Asian apes diverged. However, taking into consideration the possible short generation times in primate ancestors, it is concluded that the ancestral population size in the primate lineage was no smaller than that of extant humans. Furthermore, under the assumption of 6 million years (myr) divergence between humans and chimpanzees, the divergence time of humans from gorillas, orangutans. Old World monkeys, and New World monkeys is estimated as 7.2, 18, 34, and 65 myr ago, respectively, which are generally older than traditional estimates. Beside the intron sequences, three other data sets of orthologous sequences are used between the human and the chimpanzee comparison. The ML application to these data sets including 58,156 random BAC end sequences (BES) shows that the nucleotide substitution rate is as low as 0.6-0.8 x 10(-9) per site per year and the extent of ancestral polymorphism is 0.33-0.51%. With such a low substitution rate and short generation time, the relatively high extent of polymorphism suggests a fairly large effective population size in the ancestral lineage common to humans and chimpanzees.

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MHC diversity in Caucasians, investigated using highly heterogeneous noncoding sequence motifs at the DQB1 locus including a retroviral long terminal repeat element, and its comparison to nonhuman primate homologues.

Long terminal repeats (LTRs) are common retrovirus-related sequences spread throughout the human genome. We previously reported the human-specific integration of one LTR (DQLTR3) located 15 kb upstream of HLA DQB1. To elucidate the contribution of retroviral sequences to the variability and phylogenetic background of HLA DQB1 we investigated another LTR (DQLTR13), located 1.3 kb upstream of HLA DQB1, in German families, great apes, and Old World monkeys. Within German families, DQLTR13 presence was strongly linked to HLA DQB1*0302, *0303, and *0402 haplotypes. All other haplotypes had a low frequency or were devoid of DQLTR13. Phylogenetic analysis of DQLTR13 and adjacent nucleotide sequences in humans and non-human primates revealed a high degree of similarity and recent origin of HLA DQB1*0302, *0303, and *0402. Nevertheless, two lineages leading to DQB1*0301 and *0302 were generated by an ancient split of a DQB1*0301, *0302 progenitor. A third lineage consisting of DQB1*05/*06-related sequences may have evolved from the DQB1*0302 lineage, and a DQB1*0201-related sequence shared common ancestry with DQB1*0301. Among the human haplotypes, HLA DQB1*0201 and *0301 are linked to two different DQA1 alleles. Based on the small genetic distance of DQLTR13 as well as the adjacent sequences on these haplotypes, we suggest that a recent recombination is responsible for these associations. In the analysis of nonhuman primate species, we detected DQLTR13 in two lowland gorillas, dating the integration at at least 8 million years ago. We therefore conclude that noncoding sequences up to 1.3 kb upstream of DQB1 provide novel insight into the generation of MHC gene diversity.

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Morphological peculiarities of the deep infratemporal fossa in advanced age.

The main osseous landmarks of the border area between the infratemporal fossa and the para- and retro pharyngeal space are the sphenoidal spine and the pterygoid and styloid processes, and the styloid vagina. These landmarks, as well as some vascular anomalies, were studied in order to illustrate the variable anatomy, which is encountered in the surgical lateral transzygomatic infratemporal fossa approach. Hundred well-preserved human skull bases were examined. The deep infratemporal fossa was dissected into 54 halves of fixed cadaveric head and neck specimens of both sexes. Dry skull specimens of New and Old World monkeys, skulls of rodents, herbivora and carnivora, and computed tomograms of the head of Macaca fuscata Japonica, were also studied. In 91 of the human skull bases, the sphenoidal spine was prominent and well developed. In three skulls, the spine was absent. In four specimens, however, the spine was not sphenoidal but a part of the temporal bone, occurring in the form of a process, which emanated from the styloid vagina. In two further cases, there was unilaterally a duplicated spine; the anterior part represented a regular sphenoidal spine, while the posterior part constituted a part of the vagina of the styloid process. A complete osseous bar, arch or lamina-connecting the posterior border of the lateral lamina of the pterygoid process and the sphenoidal spine-existed in six of the human dry skulls. In ten of the human skulls examined, the breadth of the lateral lamina of the pterygoid process was greater than 10 mm; thus, the so-called pterygospinous (ps) and the pterygostyloid gates-of significance where surgical approaches are concerned-were less than 10 mm in width. Fibrous or muscular connections were also found in some cadaveric specimens between the posterior border of the lateral lamina of the pterygoid process and the sphenoidal spine: a ps ligament existed in 11 cases (20.4%) and a ps muscle in 5 cases (9.2 %), in 3 of which it inserted into both the medial wall capsule and the articular disc of the temporo-mandibular joint. Among the cadaveric specimens exhibiting ps structures was one in which an osseous ps bar occurred together with a ps muscle; in two cases a strong ps ligament was observed together with a ps muscle. The distribution pattern of the mandibular nerve was affected by the positioning of the ps bar, ligament and muscle when the latter were present. The existence of a wide ps bar was noted in all the skulls of herbivora, rodentia, carnivora, and Old World monkeys that were examined, but never in those of the New World monkeys; it is likely that, in the human, this ps bar represents a phylogenetic remnant. In the human dry skull specimens and cadaveric material, the ps ligament was found to be a reinforcement of the interpterygoid fascia, and the ps muscle to be a third head of the lateral pterygoid muscle. In two cases, the internal carotid artery exhibited a significant elongation and space-consuming tortuosity (so-called coiling behavior) in the depth of the infratemporal fossa.

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Evolution of chromosome Y in primates.

We have investigated, by fluorescence in situ hybridization (FISH), the cytogenetic evolution of the Y chromosome in primates using 17 yeast artificial chromosomes, representative of the Y-specific euchromatic region of the human chromosome Y. The FISH experiments were performed on great apes (Homo sapiens, Pan troglodytes, Gorilla gorilla and Pongo pygmaeus pygmaeus), and on two Old World monkeys species as an outgroup (Cercopitecidae Macaca fascicularis and Papio anubis). The results showed that this peculiar chromosome has undergone rapid and unconstrained evolution both in sequence content and organization.

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Entamoeba histolytica infections in captive primates.

A group based survey on the presence of Entamoeba histolytica and Entamoeba dispar using real-time PCR among 20 species of captive non-human primates was performed after diagnosis of E. histolytica dysentery in a spider monkey ( Ateles belzebuth hybridus). E. histolytica DNA was detected in three species of New World primates and in three species of Old World primates. In five of six E. histolytica isolates, it was possible to amplify the SREHP gene. They all revealed the same pattern after AluI digestion, indicating a common source of infection. E. dispar DNA was detected in two species of New World monkeys and three species of Old World monkeys. The results demonstrate that E. histolytica is capable of causing symptomatic and non-symptomatic infections in Old World and New World non-human primates. To our knowledge, this is the first report of E. histolytica sensu stricto in non-human primates after the redescription separating it from E. dispar in 1993.

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Morphological differences in Blastocystis cysts-an indication of different species?

Cyst forms of Blastocystis that show disparate morphology in relation to the previously described cysts were detected in faecal material from animal hosts. Transmission electron microscopy was performed without attempts to isolate or concentrate Blastocystis from the faecal material. Large, multinucleate cyst forms were found in faecal material from Macaca monkeys. These cyst forms measured up to approximately 15 microm in diameter and were often larger than vacuolar forms present in the same samples. Four or more nuclei were frequently seen in the cysts. Multiple individual cysts enclosed by a single fibrillar layer were found in faecal material from domestic chickens. Each individual cyst within the multiple cyst form measured approximately 3-4 microm in diameter and appeared to be uninucleate.

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