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Primate evolution of the alpha-globin gene cluster and its Alu-like repeats.

The arrangement of alpha-globin genes in Old World and New World monkeys and a prosimian, galago, has been determined by restriction mapping. Recombinant DNAs containing galago and Old World monkey alpha-globin genes have been isolated and subjected to a partial sequence determination for comparison to alpha-globin genes in human, chimpanzee and non-primate mammals. The results of this extensive structural analysis are relevant to several topics concerning the evolution of primate alpha-globin genes and Alu family repeats. All orders of higher primates (i.e. Old and New World monkeys, chimpanzee and human) have the same arrangement of alpha-globin genes. In contrast, the arrangement and correction of galago alpha-globin genes differ from those of higher primates, but are similar to those of non-primate mammals. The 5' and 3'-flanking regions of the human alpha 1 gene are orthologous to the corresponding region in galago, identifying the human alpha 2 gene as the more recently duplicated gene. The human psi alpha 1 gene is found to be inactivated after divergence of the human and galago lineages but prior to the divergence of human and monkey. Orthologous Alu family members in human and monkey DNAs indicate that the dispersion of some Alu repeats occurred prior to the divergence of these lineages. However, the Alu-like repeats of prosimian and higher primates result from entirely independent events giving rise to different repeat elements inserted at distinct genomic positions.

Animals↗

Synaptotagmin I and 1B4 are identical: implications for synaptotagmin distribution in the primate brain.

We have determined that the human cDNA sequence of the previously described primate brain mRNA species 1B4 is nearly identical (99.95% similarity) to that of human Synaptotagmin I. The apparent identity of Synaptotagmin I with 1B4, whose distribution in the brain of the monkey Cynomolgous was determined previously by Northern blot and in situ hybridization (ISH) analyses, reveals the Synaptotagmin I is differentially expressed in the primate brain. Primate Synaptotagmin I mRNA is enriched in hindbrain structures relative to forebrain structures by Northern blot analysis. By ISH analysis, primate Synaptotagmin I mRNA is highly expressed in occipital cortex and lateral geniculate (visual system components) and differentially expressed across topographic cortical boundaries between inferior and superior temporal gyrus (a polymodal zone with visual, auditory and somatosensory inputs) and between areas 17 and 18 of the visual cortex (primary and secondary visual areas). Cortical expression is also enriched in layers V and VI, which contain large pyramidal projection neurons. Synaptotagmin I's greater association with large projection neurons and with some components of visual sensory transduction could reflect a requirement of these neural components for greater synaptic activity. Synaptotagmin I expression in the primate brain is also dissimilar to Synaptotagmin I expression in rodents. Thus, variation of Synaptotagmin I expression has occurred during mammalian evolution, perhaps as a consequence of the larger size and neurotransmitter requirements of primate neurons.

Base Sequence↗

Calcium-binding protein distribution in the retina of strepsirhine and haplorhine primates.

Calcium-binding proteins are involved in numerous functional roles in the retina and are widely distributed in almost all retinal neurons. The present study aimed to characterize the distribution of the calcium-binding proteins calbindin, calretinin, parvalbumin and recoverin in relation to retinal cell types in a strepsirhine primate (mouse lemur, Microcebus) in comparison with primate species of the three main haplorhine lineages (marmoset, macaque and human), as well as a rodent (gerbil, Taterillus). The main findings show that whereas the recoverin antibody labels both rod and cone photoreceptors in all species, calbindin consistently labels cones, but not rods, in the haplorhine primates marmoset, macaque and human, but none of the photoreceptors in the mouse lemur. Marmoset and macaque also show a distinct label of cone outer segments with calretinin. Depending on the species, bipolar cells express calbindin and/or recoverin, while amacrine, horizontal and ganglion cells are labeled to varying degrees with calbindin, calretinin and parvalbumin. Haplorhine and strepsirhine primates clearly differ in the expression of calcium-binding protein expression in horizontal cells. In all haplorhine species, horizontal cells are densely labeled with parvalbumin whereas in mouse lemur horizontal cells express calbindin but not parvalbumin. Several characteristics of the calcium-binding immunostaining in the retina of the mouse lemur are similar to those observed in the rodent, and distinguish this species from the diurnal haphorhine primates. These differences may be related to adaptations of retinal structure and function to the nocturnal niche, since nocturnal strepsirhine and haphorhine (Tarsius and Aotus) primates share some features of calcium-binding expression.

Aged↗

Long-lasting in vitro hematopoiesis derived from primate embryonic stem cells.

OBJECTIVE: Induction of hematopoietic cells from human embryonic stem (ES) cells has been reported recently. However, before cells derived from human ES cells can be used in the clinic, preclinical studies using these cells in experimental primates will be necessary. Therefore, we attempted to establish a method to induce hematopoietic cells robustly and abundantly from primate ES cells. METHODS: A primate ES cell line, CMK-6, derived from the cynomolgus monkey was used in this study. We adapted a method to induce hematopoiesis from CMK-6 cells on feeder cells, and tested the effectiveness of three kinds of feeder cell lines (OP9, C2C12, and C3H10T1/2). In addition, we tested the effect of vascular endothelial growth factor (VEGF) and insulin-like growth factor-II (IGF-II) on hematopoiesis induction from CMK-6 cells. RESULTS: VEGF and IGF-II showed an extremely strong synergistic effect to induce hematopoiesis from CMK-6 cells. C3H10T1/2 cells proved to be very useful for the induction of hematopoiesis from CMK-6 cells, and the production of blood cells on C3H10T1/2 cells has been maintained as long as 5 months. During this long period, ES cell derivatives continuously produced mature blood cells, including terminally differentiated cells. CONCLUSION: We have developed an original method to produce enriched blood cells abundantly from primate ES cells for an extremely long period. This method may represent a good in vitro model for studying primate hematopoiesis and related diseases. Furthermore, our method may be useful for preclinical studies of transfusion therapy using blood cells derived from ES cells in experimental primate systems.

Animals↗

Foraging behaviour of the slender loris (Loris lydekkerianus lydekkerianus): implications for theories of primate origins.

Members of the Order Primates are characterised by a wide overlap of visual fields or optic convergence. It has been proposed that exploitation of either insects or angiosperm products in the terminal branches of trees, and the corresponding complex, three-dimensional environment associated with these foraging strategies, account for visual convergence. Although slender lorises (Loris sp.) are the most visually convergent of all the primates, very little is known about their feeding ecology. This study, carried out over 10 (1/2) months in South India, examines the feeding behaviour of L. lydekkerianus lydekkerianus in relation to hypotheses regarding visual predation of insects. Of 1238 feeding observations, 96% were of animal prey. Lorises showed an equal and overwhelming preference for terminal and middle branch feeding, using the undergrowth and trunk rarely. The type of prey caught on terminal branches (Lepidoptera, Odonata, Homoptera) differed significantly from those caught on middle branches (Hymenoptera, Coleoptera). A two-handed catch accompanied by bipedal postures was used almost exclusively on terminal branches where mobile prey was caught, whereas the more common capture technique of one-handed grab was used more often on sturdy middle branches to obtain slow moving prey. Although prey was detected with senses other than vision, vision was the key sense used upon the final strike. This study strongly supports the notion that hunting for animal prey was a key ecological determinant in selecting for visual convergence early on in primate evolution. The extreme specialisations of slender lorises, however, suggest that early primates were not dedicated faunivores and lend further support to the emerging view that both insects and fruits were probably important components of the diet of basal primates, and that exploitation of fruits may account for other key primate traits.

Animals↗

Stentless bioprosthetic heart valve research: sheep versus primate model.

BACKGROUND: The mild inflammatory response against stented bioprosthetic heart valves in the sheep model is often opposed by a more distinct response in failing human implants. With the emergence of stentless root prostheses with their significantly larger proportion of tissue interacting with the immune system of the host, a more relevant animal model than the sheep may be needed. METHODS: Valved, porcine aortic roots of 5 cm length were fixed in 0.2% glutaraldehyde and implanted in the upper descending aorta of Merino sheep (n = 5; 43+/-3 kg) and Chacma baboons (n = 5; 17+/-3 kg). After 6 weeks of tissue calcification, pannus outgrowth and inflammation were assessed by atomic absorption spectrophotometry, histologic damage scoring (0 to 3), image analysis, and transmission electron microscopy. RESULTS: The main difference between the two animal models was in aortic wall calcification (64.8+/-39.8 microg/mg in the sheep model versus 4.1+/-5.9 microg/mg in the primate model; p > 0.005). In both models, leaflet calcification was negligible (2.6+/-2.4 microg/mg in the sheep versus 2.5+/-1.9 microg/mg in the primate), and the overall extent of inflammation was comparable (1.2+/-0.8 versus 0.98+/-0.7; p = 0.18 in the sheep and the primate, respectively). Qualitatively, the sheep demonstrated a macrophage-dominated reaction whereas the inflammatory demarcation often resembled a granulocyte-dominated xenograft response in the primate. Pannus outgrowth was comparable in length (8.4+/-2.3 mm versus 9.1+/-4.3 mm proximally and 7.1+/-3.4 mm versus 7.4+/-5.1 mm distally, in the sheep and baboon, respectively; p > 0.05). CONCLUSIONS: Our results confirm the sheep as a significantly stronger calcification model for stentless aortic heart valves than the primate. Remaining antigenicity of porcine tissue as a result of incomplete cross-linking, however, elicits a distinctly stronger xenograft-type reaction in the primate model.

Animals↗

Morphological and electrophysiological properties of dissociated primate retinal cells.

Although isolated retinal cell preparations have been used widely to study retinal function in lower vertebrates, dissociated cells from primate retina have not been developed for routine physiological experiments. In this study, we demonstrated the feasibility of obtaining viable and identifiable dissociated cells from the primate retina. In addition, we characterized voltage-dependent membrane currents in each type of primate retinal cell with the whole-cell patch-clamp technique. Multiple types of ionic conductance with distinctive current profiles were recorded in various types of primate retinal neurons. Photoreceptors exhibited an inward I(H) activated by membrane hyperpolarization and an outward current activated at depolarized potentials. Two types of potassium currents (transient potassium current, I(K(A)), and delayed rectifier potassium current, I(K(V))) were recorded from bipolar cells. I(K(A)) dominated the current response in putative midget bipolar cells, and I(K(V)) was mainly associated with putative rod bipolar cells. L-type calcium currents (I(Ca)) were observed in primate bipolar cells with axon terminals, but not in axotomized bipolar cells. Large voltage-dependent sodium currents (I(Na)) were only recorded from ganglion cells. Muller cells exhibited I(K(V)) and large potassium inward rectifier current (I(K(IR))), and occasionally a small I(Na). Neurons with electrophysiological signatures of amacrine cells and horizontal cells were also studied even though their morphological features were lost during cell dissociation. By using both morphological and physiological criteria outlined in this report, it is possible to use the dissociated retinal cell preparation as an in vitro system for physiological, biochemical and pharmacological studies of the primate visual system.

Amacrine Cells↗

Locomotor behavior and control in human and non-human primates: comparisons with cats and dogs.

For many years the cat has been the accepted mammalian model for investigations on the neural control of locomotion. The results from such studies, and also similar studies on dogs, have been assumed to represent the typical mammalian condition. The primary purpose of this review is to evaluate this assumption relative to human and non-human primates. A second purpose is to acquaint investigators of mammalian locomotor behavior and control with the large amount of data available on this topic for non-human primates. The analysis shows that non-human primates are different from carnivores in footfall patterns, gaits, gait transitions, relative stride length, limb angular excursions, weight support, mechanisms of propulsion, spinal vs. supraspinal control of stepping, and possible EMG patterns. Humans exhibit more similarities with other primates than with cats or dogs, but also appear to be unique in many ways. Thus, it is clear that extrapolations of results based on cat or dog experiments may not be applicable to non-human or human primates. Furthermore, although non-human primates unquestionably make a better experimental model than cats or dogs for understanding human locomotor control mechanisms, exactly how much better remains to be determined.

Animals↗

The contribution of the MPTP-treated primate model to the development of new treatment strategies for Parkinson's disease.

Current research into Parkinson's disease (PD) is directed at developing novel agents and strategies for improved symptomatic management. The aim of this research is to provide effective and maintained symptom control throughout the course of the disease without loss of efficacy and without priming the basal ganglia for the onset of dyskinesia. To achieve these objectives, it is important to have relevant animal models of PD in which new pharmacological agents and treatment strategies can be assessed prior to clinical assessment. At present, the most effective experimental model of PD is the methyl phenyl tetrahydropyridine (MPTP)-treated primate. Primates treated with MPTP develop motor disturbances resembling those seen in idiopathic PD, including bradykinesia, rigidity and postural abnormalities. In addition, MPTP-treated primates are responsive to all commonly used antiparkinsonian agents and display treatment-associated motor complications such as dyskinesia, wearing-off and on-off, which occur during the long-term treatment of the illness. This review examines how studies conducted in MPTP-treated primates have contributed to the development of dopaminergic therapies. There is now accumulating evidence that the pulsatile manner in which short-acting agents stimulate striatal dopamine receptors is a key contributing factor to the priming of the basal ganglia for dyskinesia induction. It has been suggested that providing more continuous stimulation of dopamine receptors will avoid the development of motor complications, particularly dyskinesia. So far, the actions of all commonly used antiparkinsonian drugs assessed in MPTP-treated primates have proved to be highly predictive of drug action in PD. These primate studies have demonstrated that long-acting dopamine agonists and levodopa given in combination with a catechol-O-methyl transferase (COMT) inhibitor (to increase its relatively short half-life), induce significantly less dyskinesia than occurs with standard levodopa therapy.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Successful allotransplantation of cryopreserved tracheal grafts with preservation of the pars membranacea in nonhuman primates.

OBJECTIVE: This study was performed to confirm the feasibility of cryopreserved tracheal allotransplantation in primates, the anatomy and immunology of which are considered to be more closely related to those of humans than those of other animals. METHODS: Cryopreserved tracheal allotransplantations were performed in 3 recipient primates. In the control group fresh tracheal allotransplantations were performed in 2 primates (control A), and a tracheal allotransplantation with a simply frozen tracheal graft was performed in 1 primate (control B). Monthly bronchoscopic examinations, histologic examinations, electron microscopic examinations, and immunohistochemical investigations were performed in each of the primates. RESULTS: In the cryopreserved tracheal allotransplantation group, 3 recipient monkeys were killed on the 35th, 144th, and 387th postoperative days, respectively. All grafts were incorporated by the recipient trachea without stenosis in the cryopreserved group. In the control group 2 recipient monkeys were killed on the 93rd postoperative day (control A), and one was killed on the 84th postoperative day (control B). Severe stenosis was observed after the transplantation in all of the control monkeys. Immunologic reactions appeared to be attenuated by the cryopreservation, whereas T cell-mediated immunologic rejection (control A) and loss of cartilage viability (control B) were considered to be the causes of graft failure in the control group. CONCLUSION: The immunogenicity of the tracheal allografts was reduced by cryopreservation, and cryopreserved tracheal allotransplantation was successful in our primate model. Further investigation of cryopreserved tracheal allotransplantation with regard to proper clinical applications and the limitations of the procedure should be performed.

Animals↗

A nonhuman primate model for preclinical testing of new tuberculosis vaccines.

Nonhuman primates appear to have significant advantages over conventional laboratory animals in terms of modeling pulmonary tuberculosis for purposes of vaccine evaluation. Primates are quite susceptible to infection by the aerosol route, develop a humanlike disease, exhibit antigen-induced T lymphocyte reactivity both in vitro and in vivo, and can be protected quite effectively by bacille Calmette-Guérin vaccination. There are fewer than a dozen published studies of experimental tuberculosis in primates, and all of the available data on the response of primates to vaccination have been generated in rhesus monkeys (Macaca mulatta). There have been no modern immunologic studies of primate tuberculosis. Thus, responses to tuberculosis vaccines in primates are only minimally characterized, and much additional baseline work remains to be done before the responses to new vaccines can be placed in the proper biological context.

Animals↗

The colugo (Cynocephalus variegatus, Dermoptera): the primates' gliding sister?

Although a general agreement on the major groups of eutherian orders and their phylogenetic affiliations is emerging, the evolutionary affiliations among the members constituting these groups are still subject to debate. A prominent example is the recently published molecular evidence that challenges the long assumed monophyly of primates, displaying the colugo or flying lemur (Cynocephalus, Dermoptera) as a sister to anthropoid primates (Arnason et al. 2002 ) and positioning them after the prosimian primates (tarsiers and strepsirhines) split off. The phylogenetic analysis of the complete mitochondrial (mt) genome sequence of Cynocephalus variegatus presented in this study first appears to corroborate interpretations of primates as a paraphyletic group. However, more detailed analyses disclosed that mt nucleotide composition and consequently amino acid (AA) composition varied considerably among the species analyzed. This led us to assume that the flying lemur may be incorrectly grouped with anthropoids on the basis of similar mt nucleotide and AA compositions, rather than reflecting the true evolutionary relationship. To reanalyze the flying lemur's evolutionary association with other eutherian orders from a completely different molecular perspective, a molecular cladistic approach was applied. To this end, we determined the presence/absence pattern of transposable elements that provide a nearly homoplasy-free and copious source of molecular evolutionary markers, with well-defined character polarity. We could identify transposable elements, both on a multilocus and single-locus level, being present in all extant primate infraorders but absent in the flying lemur, thus clearly supporting the monophyly of primates by retropositional evidence.

Animals↗

The complete mitochondrial sequence of Tarsius bancanus: evidence for an extensive nucleotide compositional plasticity of primate mitochondrial DNA.

Inconsistencies between phylogenetic interpretations obtained from independent sources of molecular data occasionally hamper the recovery of the true evolutionary history of certain taxa. One prominent example concerns the primate infraordinal relationships. Phylogenetic analyses based on nuclear DNA sequences traditionally represent Tarsius as a sister group to anthropoids. In contrast, mitochondrial DNA (mtDNA) data only marginally support this affiliation or even exclude Tarsius from primates. Two possible scenarios might cause this conflict: a period of adaptive molecular evolution or a shift in the nucleotide composition of higher primate mtDNAs through directional mutation pressure. To test these options, the entire mt genome of Tarsius bancanus was sequenced and compared with mtDNA of representatives of all major primate groups and mammals. Phylogenetic reconstructions at both the amino acid (AA) and DNA level of the protein-coding genes led to faulty tree topologies depending on the algorithms used for reconstruction. We propose that these artifactual affiliations rather reflect the nucleotide compositional similarity than phylogenetic relatedness and favor the directional mutation pressure hypothesis because: (1) the overall nucleotide composition changes dramatically on the lineage leading to higher primates at both silent and nonsilent sites, and (2) a highly significant correlation exists between codon usage and the nucleotide composition at the third, silent codon position. Comparisons of mt genes with mt pseudogenes that presumably transferred to the nucleus before the directional mutation pressure took place indicate that the ancestral DNA composition is retained in the relatively fossilized mtDNA-like sequences, and that the directed acceleration of the substitution rate in higher primates is restricted to mtDNA.

Animals↗

Accelerated maturation of primate testis by xenografting into mice.

Testicular maturation and sperm production throughout the life of the male form the basis of male fertility. It is difficult to elucidate the intricate processes controlling testicular maturation and spermatogenesis in primates in vivo due to the long time span required for sexual maturation and also to the lack of accessible in vitro or in vivo models of primate spermatogenesis. Ectopic xenografting of neonatal testis tissue into mice provides an accessible model to study and manipulate the propagation and differentiation of male germ cells from immature donor animals. However, it was not clear whether this approach would be applicable to slowly maturing primates. Here we report that grafting of testis tissue from immature rhesus monkeys (Macaca mulatta) into host mice resulted in the acceleration of testicular maturation and production of fertilization-competent sperm in testis xenografts. The system reported here provides a powerful, practical approach to study timing and control of testicular maturation and regulation of primate spermatogenesis without the necessity for experimentation in primates. This approach could potentially be applied to produce fertile sperm from sexually immature individuals of rare or valuable primate species or from prepubertal boys undergoing sterilizing therapy for cancer.

Animals↗

Opposing effects of androgen and estrogen on pituitary-adrenal function in nonpregnant primates.

Maternal administration of androstenedione produces a sustained fall in maternal plasma adrenocorticotropic hormone (ACTH) concentrations in the pregnant nonhuman primate. We hypothesize a negative feedback influence on the maternal hypothalamo-pituitary-adrenal (HPA) axis by androgens in primates. This may reflect an important maternal adaptation during pregnancy in primates preventing premature induction of labor by maternal stress. However, androstenedione is precursor for placental estradiol-17beta synthesis, and infusion of androstenedione into pregnant primates elevates maternal plasma estradiol-17beta to term concentrations. Thus, it could be argued that 1) the effects attributed to androstenedione on the maternal HPA axis are mediated by estrogen rather than by androgen and 2) the negative influence of androgens may be on placental ACTH rather than, or in addition to, pituitary ACTH. To discriminate between androgenic and estrogenic effects of androstenedione on pituitary and/or placental ACTH function in primates we measured plasma ACTH, cortisol, and dehydroepiandrosterone sulfate (DHEAS) concentrations in nonpregnant baboons after treatment with either androstenedione or estradiol-17beta. Nine female baboons were studied between 14 and 22 days postpartum prior to estrous cycling. After 2 days of baseline, a continuous i.v. infusion of androstenedione (1.5 mg/kg per h in 10% intralipid, IL) was started at 0900 h and maintained for 9 days in 3 baboons. A similar protocol was carried out in another 3 baboons that received a continuous i.v. infusion of estradiol-17beta (10 microg/kg per h in 10% IL) instead of androstenedione. Three additional baboons received continuous i.v. IL vehicle alone and served as controls. Arterial blood samples (0.5 ml) for measurement of plasma hormones were taken during baseline and after 1, 3, 5, 7, and 9 days of infusion. Baseline plasma ACTH, DHEAS, and cortisol concentrations were similar among all groups. Plasma ACTH did not change during IL, increased following estradiol-17beta, and fell during androstenedione treatment. Accordingly, plasma cortisol and DHEAS concentrations were also unaltered by IL, and both steroids increased during estradiol-17beta treatment. In contrast, plasma cortisol and DHEAS remained unaltered from baseline during androstenedione treatment, despite the fall in plasma ACTH measured at this time. These data in the nonpregnant baboon 1) are consistent with negative feedback on pituitary ACTH by androgens and 2) demonstrate a positive influence on pituitary-adrenal function by estrogen in primates.

Adrenal Glands↗

The use of recombinant human bone morphogenetic protein 2 (rhBMP-2) to promote spinal fusion in a nonhuman primate anterior interbody fusion model.

STUDY DESIGN: A study on the efficacy of recombinant human bone morphogenetic protein 2 (rhBMP-2) in a nonhuman primate anterior interbody fusion model. OBJECTIVES: To investigate the efficacy of rhBMP-2 with an absorbable collagen sponge carrier to promote spinal fusion in a nonhuman primate anterior interbody fusion model. SUMMARY OF BACKGROUND DATA: RhBMP-2 is an osteoinductive growth factor capable of inducing new bone formation in vivo. Although dosage studies using rhBMP-2 have been performed on species of lower phylogenetic level, they cannot be extrapolated to the primate. Dosage studies on nonhuman primates are essential before proceeding with human primate application. METHODS: Six female adult Macaca mulatta (rhesus macaque) monkeys underwent an anterior L7-S1 interbody lumbar fusion. All six sites were assigned randomly to one of two fusion methods: 1) autogenous bone graft within a single freeze-dried smooth cortical dowel allograft cylinder (control) or 2) rhBMP-2-soaked absorbable collagen sponges within a single freeze-dried smooth cortical dowel allograft cylinder also soaked in rhBMP-2. The animals underwent a baseline computed tomography scan followed by 3- and 6-month postoperation scans. Anteroposterior and lateral radiographs of the lumbosacral spine were performed monthly. After the monkeys were killed, the lumbar spine fusion sites were evaluated. Histologic evaluation of all fusion sites was performed. RESULTS: The three monkeys receiving rhBMP-2-soaked collagen sponges with a freeze-dried allograft demonstrated radiographic signs of fusion as early as 8 weeks. The control animals were slower to reveal new bone formation. The computed tomography scans revealed extensive fusion of the L7-S1 lumbar vertebrae in the group with rhBMP-2. A pseudarthrosis was present in two of the control animals. CONCLUSIONS: This study was able to document the efficacy of rhBMP-2 with an absorbable collagen sponge carrier and a cortical dowel allograft to promote anterior interbody fusion in a nonhuman primate model at a dose of 0.4 mg per implant site (1.5 mg/mL concentration). The late of new bone formation and fusion with the use of rhBMP-2 and cortical dowel allograft appears to be far superior to that of autogenous cancellous iliac crest graft with cortical dowel allograft.

Animals↗

Porcine hematopoiesis on primate stroma in long-term cultures: enhanced growth with neutralizing tumor necrosis factor-alpha and tumor growth factor-beta antibodies.

BACKGROUND: Donor hematopoiesis is at a competitive disadvantage when bone marrow transplantation is across species barriers. This could present major limitations to xenogeneic stem cell transplantation as an approach to tolerance induction. An in vitro model of xenogeneic engraftment was established to identify inhibitors of porcine hematopoiesis in a primate environment. METHODS: Porcine bone marrow cells (BMC), in the presence or absence of primate CD34+ positive cells, were cultured for 4-6 weeks on primate stroma with porcine cytokines. Cellularity and growth of colony-forming cells were indicators of hematopoietic growth. Effects of soluble factors were determined by using Transwell inserts to separate porcine cells from stroma. Neutralizing antibodies for human transforming growth factor-beta (TGF-beta) and tumor necrosis factor-alpha (TNF-alpha) were added to cultures. RESULTS: Porcine hematopoiesis can be maintained in long-term cultures on primate stroma with pig cytokines. Adding BMC to the stroma below Transwell-containing porcine cells dramatically inhibited porcine hematopoiesis, showing an inhibitory role for soluble factors. Neutralizing antibodies against TNF-alpha or TGF-beta caused a modest enhancement of porcine hematopoiesis; however, the combination of both led to a substantial increase. Inhibitory effects of these cytokines were confirmed by adding TNF-alpha and TGF-beta to porcine cultures. CONCLUSIONS: Porcine cells may be more sensitive to inhibitory effects of TNF-alpha and TGF-beta than primate cells and are at a disadvantage when in a primate environment. Potential implications of this observation are discussed in the context of establishing specific immune tolerance via mixed chimerism to facilitate xenotransplantation.

Animals↗

Visual field representation in striate and prestriate cortices of a prosimian primate (Galago garnetti).

Microelectrode mapping techniques were used to study the visuotopic organization of the first and second visual areas (V1 and V2, respectively) in anesthetized Galago garnetti, alorisiform prosimian primate. 1) V1 occupies approximately 200 mm2 of cortex, and is pear shaped, rather than elliptical as in simian primates. Neurons in V1 form a continuous (1st-order) representation of the visual field, with the vertical meridian forming most of its perimeter. The representation of the horizontal meridian divides V1 into nearly equal sectors representing the upper quadrant ventrally, and the lower quadrant dorsally. 2) The emphasis on representation of central vision is less marked in Galago than in simian primates, both diurnal and nocturnal. The decay of cortical magnification factor with increasing eccentricity is almost exactly counterbalanced by an increase in average receptive field size, such that a point anywhere in the visual field is represented by a compartment of similar diameter in V1. 3) Although most of the cortex surrounding V1 corresponds to V2, one-quarter of the perimeter of V1 is formed by agranular cortex within the rostral calcarine sulcus, including area prostriata. Although under our recording conditions virtually every recording site in V2 yielded visually responsive cells, only a minority of those in area prostriata revealed such responses. 4) V2 forms a cortical belt of variable width, being narrowest (approximately 1 mm) in the representation of the area centralis and widest (2.5-3 mm) in the representation of the midperiphery (>20 degrees eccentricity) of the visual field. V2 forms a second-order representation of the visual field, with the area centralis being represented laterally and the visual field periphery medially, near the calcarine sulcus. Unlike in simians, the line of field discontinuity in Galago V2 does not exactly coincide with the horizontal meridian: a portion of the lower quadrant immediately adjacent to the horizontal meridian is represented at the rostral border of ventral V2, instead of in dorsal V2. Despite the absence of cytochrome oxidase stripes, the visual field map in Galago V2 resembles the ones described in simians in that the magnification factor is anisotropic. 5) Receptive field progressions in cortex rostral to dorsal V2 suggest the presence of a homologue of the dorsomedial area, including representations of both quadrants of the visual field. These results indicate that many aspects of organization of V1 and V2 in simian primates are shared with lorisiform prosimians, and are therefore likely to have been present in the last common ancestor of living primates. However, some aspects of organization of the caudal visual areas in Galago are intermediate between nonprimates and simian primates, reflecting either an intermediate stage of differentiation or adaptations to a nocturnal niche. These include the shape and the small size of V1 and V2, the modest degree of emphasis on central visual field representation, and the relatively large area prostriata.

Animals↗