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Genetically modified adenovirus vector containing an RGD peptide in the HI loop of the fiber knob improves gene transfer to nonhuman primate isolated pancreatic islets.

The ability to transfer immunoregulatory, cytoprotective, or antiapoptotic genes into pancreatic islets (PIs) may allow enhanced post-transplantation survival. The available gene transfer vectors differ greatly in their ability to infect and express genes in different cell types. One limitation associated with the use of viral vectors is related to the virus reliance on the presence of its primary binding site. Tropism of the viral vectors can be altered using retargeting strategies. Results on phage biopanning proved that the RGD motif has in vivo targeting capabilities. This motif interacts especially with cellular integrins of the alphavbeta3 and alphavbeta5 types, highly expressed on pancreatic islets. In this report, we have explored the utility of a retargeted adenovirus vector (Ad) containing an RGD motif in the HI loop of the fiber knob in order to improve the infection efficiency to intact isolated nonhuman primate PIs and reduce toxicity after the genetic modification. Nonhuman primate Pis were isolated by a semi-automated technique. Steptozotocin-induced diabetic mice with severe combined immunodeficiency disease (SCID) were used as recipients. A recombinant Ad containing a heterologous RGD peptide and expressing luciferase (AdRGDLuc) or green fluorescent protein (AdRGDGFP) were generated in our laboratory. Similar Ads without the RGD peptide were used as a control (AdLuc and AdGFP). Higher transfection efficiency was demonstrated using AdRGDGFP compared with AdGFP (>80% of the islet cells were infected at 10 particle-forming units (pfu)/cell using AdRGDGFP vs. 7% after infection with AdGFP).More than 90% of the infected cells were insulin-producing cells. Significantly higher transgene expression was demonstrated after infection with AdRGDLuc compared with AdLuc at different titers. Analysis of the glucose-stimulated insulin response demonstrated better performance of PI transfected with AdRGDLuc at low titers (10 pfu/cell in order to achieve > 80% transfection efficiency) compared with AdLuc at high titers. Finally, long-term euglycemia (>250d) was observed in 89% of the animals that received PI infected with AdRGDLuc compared with none of the animals that received PI infected with AdLuc. The present study provides new information about the possibility of tropism modification of Ad vectors to increase the transfection efficiency and transgene expression to isolated PI. Incorporation of the RGD sequence in the HI loop of the fiber knob allows highly efficient transfection efficiency to nonhuman primate insulin-producing cells and adequate long-term function of the p-cell after transplantation.

Adenoviridae↗

Primate cognition and the bared-teeth display: a reevaluation of the concept of formal dominance.

The cognitive implications of the phenomenon of dominance in nonhuman primates and other animals have been poorly addressed. This article shows that the concept of formal dominance currently used in the primatological literature is inherently dependent on the assumption that primates are capable of attribution of knowledge to other individuals. Although this assumption has never been made explicitly, without it the concept of formal dominance is virtually indistinguishable from other, more traditional views of dominance. Recent studies have failed to demonstrate attribution of knowledge in nonhuman primates, thus questioning the validity of formal dominance. The concept of formal dominance was originally proposed to account for a discrepancy between dominance hierarchies based on aggressive behavior and dominance hierarchies based on submissive signals. This article shows that such a discrepancy can be accounted for without invoking complex cognitive processes such as attribution of knowledge.

Agonistic Behavior↗

A meta-analysis of primate hand preferences, particularly for reaching.

P. F. MacNeilage, M. G. Studdert-Kennedy, and B. Lindblom (1987) proposed a progression for handedness in primates that was supposed to account for the evolution of a right bias in human handedness. To test this proposal, the authors performed meta-analyses on 62 studies that provided individual data (representing 31 species: 9 prosimians, 6 New World monkeys, 10 Old World monkeys, 2 lesser apes, and 4 greater apes), of the 118 studies of primate handedness published since 1987. Although evidence of a population-level left-handed bias for prosimians and Old World monkeys supports P. F. MacNeilage et al., the data from apes, New World monkeys, and individual species of prosimians and New World monkeys do not. Something other than primate handedness may have been the evolutionary precursor of the right bias in hand-use distribution among hominids.

Animals↗

A common mammalian plan of accessory optic system organization revealed in all primates.

The accessory optic system (AOS), which was described as early as 1870 by Gudden, constitutes a distinct midbrain visual pathway in all classes of vertebrates. In non-primate mammals, retinal fibres of this system project to a set of three nuclei: the dorsal (DTN), the lateral (LTN) and the medial (MTN) terminal nuclei. Whereas all AOS cells respond to the slow motion of large visual stimuli, the neurons are tuned to complementary directions of movement: horizontal temporo-nasal direction for the DTN, vertical up and down for the LTN and vertical down for the MTN. It has thus been suggested that these nuclei establish a system of retinal coordinates for the detection of whole field motion. As the AOS provides direct and indirect pathways to both oculomotor and vestibular structures, each of these nuclei is thought to be an essential link in the co-ordination of eye and head movements in relation to movement within the visual-field. One problem for the generalization of this theory is that the medial terminal nucleus has never been found in primates. In this report we establish both the existence of this nucleus and its afferent input from the retina in all major groups of primates (prosimians, New and Old World monkeys and apes), indicating a common anatomical plan of organization of the AOS in mammals.

Animals↗

Dispersal and the sex ratio at birth in primates.

The females of many species of primates settle for life within the home ranges of their mothers, whereas males disperse as immatures. According to the theory of sex allocation, the costs incurred by mothers through local competition for resources with their philopatric daughters should favour the evolution of male-biased sex ratios at birth. I report here two tests of this hypothesis based on data from 15 genera of primates. First, I show that the intensity of competition for resources within kin groups is strongly and positively correlated with sex ratios at birth. Second, I show that sex ratios at birth are higher in genera with female-biased philopatry than in genera in which philopatry is not female-biased. These analyses suggest that local resource competition among kin powerfully influences the evolution of sex ratios in primates.

Animals↗

First skulls of the early Eocene primate Shoshonius cooperi and the anthropoid-tarsier dichotomy.

The phylogenetic relationships of living tarsiers and extinct omomyid primates are critical for deciphering the origin and relationships of primate higher taxa, particularly anthropoids. Three competing phylogenetic hypotheses are: (1) tarsiers are most closely related to early Cenozoic Omomyidae, particularly genera such as Necrolemur from the late Eocene of Europe; (2) tarsiers share a more recent common ancestry with anthropoids than they do with any known omomyid; (3) tarsiers and/or omomyids are most closely related to strepsirhines. The anatomy of four skulls of the early Eocene omomyid Shoshonius cooperi--the first cranial material recovered for this genus--strongly suggests that Shoshonius shares a more recent common ancestry with Tarsius than do either anthropoids or other Eocene omomyids for which cranial anatomy is known. If the primate suborder Haplorhini (anthropoids, omomyids, tarsiids) is monophyletic, the phylogenetic position of Shoshonius requires that anthropoids and Tarsius diverged by at least the early Eocene, some 15 million years before the first appearance of anthropoids in the fossil record.

Animals↗

Ecological importance of trichromatic vision to primates.

Trichromatic colour vision, characterized by three retinal photopigments tuned to peak wavelengths of approximately 430 nm, approximately 535 nm and approximately 562 nm (refs 1, 2), has evolved convergently in catarrhine primates and one genus of New World monkey, the howlers (genus Alouatta). This uniform capacity to discriminate red-green colours, which is not found in other mammals, has been proposed as advantageous for the long-range detection of either ripe fruits or young leaves (which frequently flush red in the tropics) against a background of mature foliage. Here we show that four trichromatic primate species in Kibale Forest, Uganda, eat leaves that are colour discriminated only by red-greenness, a colour axis correlated with high protein levels and low toughness. Despite their divergent digestive systems, these primates have no significant interspecific differences in leaf colour selection. In contrast, eaten fruits were generally discriminated from mature leaves on both red-green and yellow-blue channels and also by their luminance, with a significant difference between chimpanzees and monkeys in fruit colour choice. Our results implicate leaf consumption, a critical food resource when fruit is scarce, as having unique value in maintaining trichromacy in catarrhines.

Adaptation, Biological↗

A new Late Eocene anthropoid primate from Thailand.

The fossil record of anthropoid primates from the Middle Eocene of South Asia is so far restricted to two genera (Pondaungia cotteri Pilgrim, 1937 and Amphipithecus mogaungensis Colbert, 1937 from the Eocene Pondaung deposits of Burma) whose anthropoid status and phylogenetic position have long been under debate because they represent the oldest highly derived fossil primates of anthropoid grade. Moreover, several new African taxa, some of which are even older, have been recently included in the suborder Anthropoidea, suggesting an African origin for this group. Conversely, new fossil primates recently discovered in China (Eosimias) have been related to the most primitive representatives of Anthropoidea, alternatively suggesting an Asian origin and a probable Asian radiation centre. We report here the discovery of a new anthropoid from the Thai Late Eocene locality of Krabi, which displays several additional anthropoid characters with regard to those of the Eocene Burmese genera. This species, which is about the size of the Fayum Aegyptopithecus, can be related to the Burmese forms, and it further provides strong additional evidence for a southeast Asian evolutionary centre for anthropoids.

Animals↗

Enrichment devices for nonhuman primates.

There has been in recent years a substantial increase in the variety of enhancement devices available for nonhuman primates in captivity, and the task of properly outfitting a housing unit can be daunting. Researchers continue to investigate the specific impact and importance of environmental enrichment, but it is generally accepted that increasing the complexity of the environment for the mental and physical stimulation of nonhuman primates is beneficial to their health and contentment, and enrichment is now a standard component of primate husbandry.

Animal Welfare↗

Ancient co-speciation of simian foamy viruses and primates.

Although parasite-host co-speciation is a long-held hypothesis, convincing evidence for long-term co-speciation remains elusive, largely because of small numbers of hosts and parasites studied and uncertainty over rates of evolutionary change. Co-speciation is especially rare in RNA viruses, in which cross-species transfer is the dominant mode of evolution. Simian foamy viruses (SFVs) are ubiquitous, non-pathogenic retroviruses that infect all primates. Here we test the co-speciation hypothesis in SFVs and their primate hosts by comparing the phylogenies of SFV polymerase and mitochondrial cytochrome oxidase subunit II from African and Asian monkeys and apes. The phylogenetic trees were remarkably congruent in both branching order and divergence times, strongly supporting co-speciation. Molecular clock calibrations revealed an extremely low rate of SFV evolution, 1.7 x 10(-8) substitutions per site per year, making it the slowest-evolving RNA virus documented so far. These results indicate that SFVs might have co-speciated with Old World primates for at least 30 million years, making them the oldest known vertebrate RNA viruses.

Animals↗

Evolution of the beta defensin 2 gene in primates.

With the aim of further investigating the molecular evolution of beta defensin genes, after having analysed beta defensin 1 (DEFB1) in humans and several nonhuman primate species, we have studied the evolution of the beta defensin 2 gene (DEFB2), which codifies for a peptide with antimicrobial and chemoattractant activity, in humans and 16 primate species. We have found evidence of positive selection during the evolution of orthologous DEFB2 genes at two points on a phylogenetic tree relating these primates: during the divergence of the platyrrhines from the catarrhines and during the divergence of the Cercopithecidae from the Hylobatidae, Great Apes and humans. Furthermore, amino acid variations in Old World Monkeys seem to centre either on residues that are involved in oligomerisation in the human molecule, or that are conserved (40-80%) in beta-defensins in general. It is thus likely that these variations affect the biological function of the molecules and suggest that their synthesis and functional analysis might reveal interesting new information as to their role in innate immunity.

Amino Acid Sequence↗

Antibody-mediated lung endothelium targeting: in vivo model on primates.

We have recently provided evidence that angiotensin-converting enzyme (ACE) is a rational target and anti-ACE monoclonal antibodies (mAbs) are suitable molecules for directing gene/drug delivery into the pulmonary endothelium of rodents. As a step towards gene therapy clinical trials using this approach, the present study evaluated the potential of anti-ACE mAbs for in vivo lung endothelium targeting in 10 species of primates. Cross-reactivity of 10 distinct mAbs directed to human ACE with ACE from baboon, macaques, cercopithecus and chimpanzee revealed that the highest binding with ACE from baboon and macaques was with mAb i2H5, from chimpanzee - mAb 9B9, and from human - 9B9 and i2H5. Thereafter, in vivo biodistribution of mAbs i2H5 and 9B9 was estimated in Macaca arctoides. MAb i2H5, which binds to macaque ACE with substantially higher affinity than mAb 9B9, also more effectively accumulates in their lungs than mAb 9B9. Immunospecificity of lung accumulation (mAb/control IgG ratio) was 37 for i2H5 and 0.5 for 9B9. Lung selectivity of i2H5 uptake (lung/blood ratio) was around 10. Therefore mAb i2H5 may be useful for in vivo lung targeting in non-human primates, whereas 9B9 may be most useful in primates that are closer to humans (chimpanzee). A combination of these two mAbs may be particularly useful for human clinical trials of gene/drug therapy for lung disorders such as pulmonary hypertension and lung metastases.

Animals↗

Genetic marking as an approach to studying in vivo hematopoiesis: progress in the non-human primate model.

Retroviral insertion site analysis following transplantation of marked hematopoietic stem cells (HSCs) is a powerful method for studying hematopoiesis in vivo. High-level gene transfer efficiency was achieved in murine models in the late 1980s, but early human gene transfer protocols into hematopoietic stem and progenitor cells using the murine methodology showed consistently poor results. The utility of non-human primates as pre-clinical models has since become apparent. Modifications in retroviral transduction conditions have resulted in stable long-term gene transfer efficiency as high as 15-20% to primitive repopulating cells in non-human primate models. This has permitted, for the first time in a large animal model, tracking of individual stem and progenitor cell clones via insertion site analysis, an advantage over competitive transplantation studies, which cannot firmly evaluate the number or life span of individual clones contributing to hematopoiesis. Retroviral tracking studies in mice suggest that stable hematopoiesis may be dominated by a small number of clones, but these studies have been limited by insensitive detection methods, low numbers of transplanted stem cells, and limited life span of immunodeficient mice. Autologous transplantation studies in non-human primates have just begun and have the potential to shed light on controversial issues such as the number of clones contributing to stable hematopoiesis, clonal succession, and lineage commitment, as well as the effect of clinically relevant manipulations such as cytokines, chemotherapy, and radiation on hematopoiesis. These approaches will have significant impact in studying various aspects of stem cell biology including the phenomenon of stem cell plasticity.

Animals↗

Primate encephalization and intelligence.

The amount of brain mass exceeding that related to an animal's total body mass is called encephalization. And quantifying encephalization has been argued to be directly related to an animal's level of intelligence. In this study, the legitimacy of the encephalization hypothesis was tested for encephalization slopes 0.28, 0.67, and 0.75 in Snell's equation of simple allometry by determining the intersexual encephalization similarity for humans and 18 other primate species and by comparing the encephalization quotients for humans and six other primate species against the learning ability evaluations for their intelligence. Results suggest that the most accurate means for quantifying the encephalization of humans and other adult primate species requires the use of Lapicque's universal exponent of 0.28 in Snell's equation of simple allometry. Since Lapicque's slope was derived from various vertebrate groups, this equation may potentially be universally applicable for determining relative adult vertebrate encephalization and intelligence.

Animals↗

Variations in adrenal androgen production among (nonhuman) primates.

The synthesis and secretion of the adrenal androgens dehydroepiandrosterone (DHEA) and its sulfate (DS) is a phenomenon apparently unique to humans and nonhuman primates. It occurs at three life stages: in utero from the fetal zone (FZ) cells of the developing adrenal cortex, during adolescence with the onset of adrenarche and the development of the zona reticularis (ZR), and in ever decreasing amounts from the ZR with aging (adrenal senescence). Insufficient data exist to know if any single nonhuman primate exactly mirrors human adrenal androgen secretion through all three life stages, and detailed morphological, biochemical, and endocrinologic studies are required to do so. Androgen synthesis requires that cells express three key enzymes, 17alpha-hydroxylase/17,20-lyase cytochrome P450 (P450c17), nicotinamide-adenine dinucleotide phosphate (NADPH)-cytochrome P450 oxidoreductase (CPR), and cytochrome b5, and that they do not express 3beta-hydroxysteroid dehydrogenase (3beta-HSD). Cytochrome b5 has emerged as a particularly useful marker of androgen synthetic potential. Although a reliable index of the rate of adrenal androgen secretion, DS concentrations may not accurately reflect total adrenal androgen output because rates and routes of androgen metabolism may vary greatly among species. Based on the very limited available data, the most promising nonhuman primate models are marmosets for the human FZ, chimpanzees for human adrenarche, and macaques and baboons for mature ZR function that declines with senescence.

Adrenal Glands↗

Structural and functional maturation of the developing primate brain.

Descriptive studies have established that the developmental events responsible for the assembly of neural systems and circuitry are conserved across mammalian species. However, primates are unique regarding the time during which histogenesis occurs and the extended postnatal period during which myelination of pathways and circuitry formation occur and are then subsequently modified, particularly in the cerebral cortex. As in lower mammals, the framework for subcortical-cortical connectivity in primates is established before midgestation and already begins to remodel before birth. Association systems, responsible for modulating intracortical circuits that integrate information across functional domains, also form before birth, but their growth and reorganization extend into puberty. There are substantial differences across species in the patterns of development of specific neurochemical systems. The complexity is even greater when considering that the development of any particular cellular component may differ among cortical areas in the same primate species. Developmental and behavioral neurobiologists, psychologists, and pediatricians are challenged with understanding how functional maturation relates to the evolving anatomical organization of the human brain during childhood, and moreover, how genetic and environmental perturbations affect the adaptive changes exhibited by neural circuits in response to developmental disruption.

Animals↗

Embryo implantation and embryonic stem cell development in primates.

The endocrine dialogue that results in implantation and the successful establishment of pregnancy in primates relies on embryonic secretion of chorionic gonadotrophin (CG). This hormone is a signal of embryo viability and capacity to support the corpus luteum. The expression of CG is apparently restricted to primates. Active or passive immunization of marmoset monkeys against the beta subunit of CG prevented implantation and early pregnancy, without disrupting the ovarian cycle. Studies of individual embryos cultured in vitro showed that CG is secreted at low levels by the blastocyst from before attachment, with secretion increasing exponentially after attachment. Gonadotrophin releasing hormone (GnRH) was also secreted, from mid-blastocyst stages, before the detection of CG. The secretion of GnRH by the embryo continued through the attachment and outgrowth stages of embryonic differentiation in vitro. The hypothetical role of GnRH in regulating CG release during implantation was tested in recently completed experiments. Individual embryos cultured with GnRH, or with agonist or antagonist to GnRH, showed significant variations in their secretion of CG and in their survival in culture, suggesting a causal relationship between these hormones. Embryos cultured with natural GnRH showed enhanced growth and development. Embryonic stem cells, from the inner cell mass of marmoset and rhesus monkeys, were the first primate embryonic stem cells to be isolated and characterized, enabling the subsequent isolation of human embryonic stem cells.

Animals↗

Flow cytometric sorting of frozen-thawed spermatozoa in sheep and non-human primates.

Research was conducted in sheep to determine an effective preparation method for high-purity sorting of frozen-thawed spermatozoa. The efficacy of sorting frozen-thawed spermatozoa was then investigated in several non-human primate species. An aliquot of each ejaculate (three rams, three ejaculates per ram) was processed as a fresh control (FRESH). Frozen spermatozoa were thawed and prepared for sorting by no further processing (FT-NEAT), washing (FT-WASH) or gradient centrifugation (FT-GRADIENT) and evaluated for motility at 1 h post-staining and motility and acrosomal status at 0 and 4 h post-sorting. Samples were analysed using a high-speed cell sorter. High levels of purity for X- and Y-enriched samples were achieved for all treatments (85-92%). The percentage of motile spermatozoa before sorting was lower (P < 0.05) for frozen-thawed samples (FT-NEAT: 32.7 +/- 2.5%; FT-WASH: 32.2 +/- 3.3%; FT-GRADIENT: 73.9 +/- 3.7%) compared with FRESH (83.3 +/- 1.2%). Post-sorting, the percentage of motile spermatozoa before and after incubation for FT-NEAT (60.0 +/- 5.1% and 27.2 +/- 6.1% for 0 and 4 h, respectively) was lower than that for FRESH (87.8 +/- 0.9% and 83.3 +/- 1.2% for 0 and 4 h, respectively; P < 0.05), FT-WASH (80.0 +/- 2.4% and 71.7 +/- 3.6% for 0 and 4 h, respectively; P < 0.05) and FT-GRADIENT (84.4 +/- 1.3% and 77.2 +/- 1.7% for 0 and 4 h, respectively; P < 0.05). Vanguard sperm migration distance through artificial cervical mucus was lower (P < 0.05) for FT-NEAT (17.7 +/- 1.7 mm) compared with FT-WASH (29.1 +/- 3.8 mm) and FT-GRADIENT (28.4 +/- 2.0 mm) and similar (P < 0.05) to FRESH (23.7 +/- 1.8 mm). Sample preparation using a modified wash method enabled high-purity sorting (range 86-97% purity) of frozen-thawed epididymal spermatozoa in the baboon (Papio hamadryas), common marmoset (Callithrix jacchus) and common chimpanzee (Pan troglodytes). For all non-human primate species, sorted spermatozoa were progressively motile (marmoset: 20.5 +/- 5.5%; baboon: 37.5 +/- 2.5%; chimpanzee: 73.0 +/- 2.0%), acrosome intact (marmoset: 68.5 +/- 7.5%; baboon: 89.5 +/- 1.5%; chimpanzee: 84.0 +/- 1.0%) and able to penetrate an artificial cervical mucus. In summary, high-purity sorting of frozen-thawed ram and non-human primate spermatozoa with recovery of progressively motile, acrosome-intact spermatozoa was possible after processing to remove cryodiluent.

Animals↗