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Pathologic features of vascular endothelial growth factor-induced retinopathy in the nonhuman primate.

PURPOSE: Vascular endothelial growth factor (VEGF) is a potent ischemia-upregulated angiogenic protein that has been implicated in diabetic retinopathy. Intravitreal VEGF injections have not previously been shown to produce preretinal neovascularization. The purpose of this study was to further characterize the angiopathic changes that occur after intravitreal injections in a nonhuman primate and determine if preretinal neovascularization develops. DESIGN: Experimental animal study. METHODS: Vascular endothelial growth factor 165 was injected into the eyes of normal cynomolgus monkeys at regular intervals. As a control, normal eyes were injected with phosphate buffered saline. Color photography and fluorescein angiography were performed at regular intervals. The retinas were incubated for adenosine diphosphatase (ADPase) activity to visualize retinal vessels. The retinas were flat-embedded and areas of potential preretinal neovascularization were identified en bloc and serially sectioned. RESULTS: Areas of capillary nonperfusion and vessel dilation and tortuousity were seen by angiography. In serial sections, the nonperfused areas were found to be associated with endothelial cell hyperplasia in vessel lumens. Preretinal neovascularization originating only from superficial veins and venules was observed throughout peripheral retina, but was not seen in the posterior pole. Lacunae-like veins were subdivided by the process of intussusception and endothelial cell bridging. Arterioles demonstrated endothelial cell hyperplasia and microaneurysms. CONCLUSION: Intraocular injections of VEGF were sufficient to produce preretinal neovascularization in the nonhuman primate. Most vasculopathic structures were associated with endothelial cell hyperplasia. These results demonstrate that VEGF alone can produce many features of both nonproliferative and proliferative diabetic retinopathy including the previously undescribed development of preretinal neovascularization. This well-characterized VEGF-induced primate model of retinal neovascularization may be useful as a means of testing new treatments for retinal neovascularization.

Animals↗

A nonhuman primate model for the selective elimination of CD8+ lymphocytes using a mouse-human chimeric monoclonal antibody.

Nonhuman primates provide valuable animal models for human diseases. However, studies assessing the role of cell-mediated immune responses have been difficult to perform in nonhuman primates. We have shown that CD8+ lymphocyte-mediated immunity in rhesus monkeys can be selectively eliminated using the mouse-human chimeric anti-CD8 monoclonal antibody cM-T807. In vitro, this antibody completely blocked antigen-specific expansion of cytotoxic T cells and decreased major histocompatibility complex class I-restricted, antigen-specific lysis of target cells but did not mediate complement-dependent cell lysis. In vivo administration of cM-T807 in rhesus monkeys resulted in near total depletion of CD8+ T cells from the blood and lymph nodes for up to 6 weeks. This depletion was not solely complement-dependent and persisted longer in adults than in juveniles. Preservation of B cell and CD4+ T cell function in monkeys depleted of CD8+ lymphocytes was demonstrated by their ability to develop humoral immune responses to the administered chimeric monoclonal antibody. Furthermore, during CD8+ lymphocyte depletion, monkeys developed delayed-type hypersensitivity reactions comprised only of CD4+ T cells but not CD8+ T cells. This CD8+ lymphocyte depletion model should prove useful in defining the role of cell-mediated immune responses in controlling infectious diseases in nonhuman primates.

Animals↗

Modified technique for heterotopic heart transplantation in small primates.

Cardiac transplantation in small primates represents a unique model for investigating both xenograft and allograft rejection. This report describes our technique for heterotopic transplantation of cardiac grafts into the retroperitoneal iliac vessels of newborn baboons and small primates. Small primates tolerate this position better than either cervical or abdominal placement.

Animals↗

Variable foraging demand rearing: sustained elevations in cisternal cerebrospinal fluid corticotropin-releasing factor concentrations in adult primates.

BACKGROUND: The authors previously reported elevated cerebrospinal fluid (CSF) corticotropin-releasing factor (CRF) concentrations in juvenile primates nursed by mothers undergoing experimentally imposed unpredictable foraging conditions in comparison to normally reared controls. The purpose of the present study was to determine if these changes would endure into young adulthood. METHODS: Cisternal CSF samples were obtained from those unpredictably reared young adult primates who had been previously studied as juveniles and age-matched ad libitum normally reared controls. Samples were assayed for CSF CRF. RESULTS: Concentrations of CSF CRF were significantly elevated in the unpredictably reared sample in comparison to the ad libitum-reared control group. A significant positive correlation was noted between juvenile and young adult CSF CRF values within the unpredictably reared cohort. CONCLUSIONS: Disturbances of maternal-infant attachment processes have an enduring impact on primate CRF function into young adulthood. The CRF elevations following unpredictable maternal foraging conditions appear traitlike in nature.

Age Factors↗

Pharmacological characterization of the vesamicol analogue (+)-[(125)I]MIBT in primate brain.

The vesamicol analogue, meta-[(125)I]iodobenzyltrozamicol [(+)-[(125)I]MIBT] was evaluated as a probe for the in vitro labeling of the vesicular acetylcholine transporter in primate brain. In the striatum, (+)-[(125)I]MIBT bound a single high-affinity site with a Kd value of 4.4 +/- 0.7 nM. Competition for (+)-[(125)I]MIBT binding to the striatum by a group of vesamicol analogues displayed a pharmacological profile similar to the rank order of potency previously observed for the vesicular acetylcholine transporter on Torpedo synaptic vesicles. High-affinity binding of (+)-[(125)I]MIBT in the occipital cortex was characterized by a Kd value of 4.6 +/- 1.1 nM. However, the rank order of potency for inhibition of (+)-[(125)I]MIBT binding to the occipital cortex by the same test compounds differed from that observed in the striatum. The results suggest that (+)-[(125)I]MIBT is a reliable probe of the vesicular acetylcholine transporter in primate striatum, but its binding in primate occipital cortex is more complex.

Acetylcholine↗

Baseline characteristics of the transient pattern electroretinogram in non-human primates: inter-ocular and inter-session variability.

This study assessed the inter-ocular and inter-session variability of the transient pattern electroretinogram (PERG) in a group of non-human primates. The transient PERG was measured both eyes of 29 non-human primates, and again after three months in 23 eyes of 23 of these animals. Signals were elicited using a contrast (90%, 75 cdm(-2)) reversing (5 reversals sec(-1)) checkerboard pattern (0.56 cpd). PERGs were also measured for stimuli of varied spatial frequency (n=8, 0.07-2.22 cpd), contrast (n=4, 20-100%), mean luminance (n=4, 4.7-75 cdm(-2)) and defocus (n=5, +1, +2, +3 diopters). The inter-eye and inter-session limits-of-agreement (LOA; 95%) were determined for each PERG parameter. Variability was also compared with previous studies using the coefficient-of-variability (COV). Pharmacological blockade of the inner retinal contributions to the PERG measured under these conditions was conducted in one animal using intravitreal injection of tetrodotoxin (approximately 6 microM) and N-methyl-D-aspartic acid (approximately 6 microM). The N95 component of the primate transient PERG showed spatial tuning, with a peak between 0.14 and 0.28cpd. This spatial tuning was not as apparent for the P50 component. A linear relationship between P50 and N95 amplitude was found with contrast and mean luminance. Both components were attenuated with the introduction of +2 diopters or more of defocus. The inter-session COV for the P50 and N95 components were 23.8 and 19.2%, respectively, while the LOA were 58 and 46%, respectively. The N95:P50 ratio had smaller inter-session variability, was robust to changes in contrast, mean luminance and defocus, and was effective for characterization of inner-retinal dysfunction after pharmacologic block.

Animals↗

Use of the progestin challenge test in nonhuman primates (Macaca fascicularis).

OBJECTIVE: To determine whether the progestin challenge test (PCT) would provide a reliable, noninvasive indicator of endometrial stimulation in nonhuman primates. DESIGN; Randomized, 2x2, crossover study. SETTING; Nonhuman primates (Macaca fascicularis) in an academic research environment. PATIENT(S): Adult, surgically postmenopausal, female cynomolgous macaques (n = 27) were studied. INTERVENTION(S): Females were randomly assigned to receive estradiol (n = 14; 0.028 mg/kg body weight) or vehicle (n = 13) daily. All animals were administered two PCTs in a crossover study design using two doses (0.28 mg/kg or 0.56 mg/kg body weight) of medroxyprogesterone acetate (MPA). MAIN OUTCOME MEASURE(S): Incidence and severity of withdrawal bleeding and serum estradiol (E(2)) and progesterone (P(4)) levels were evaluated. RESULT(S): Estradiol treatment resulted in endometrial "withdrawal" bleeding in all but one instance. Females receiving daily doses of E(2) exhibited a significantly greater (P<.01) incidence, severity, and duration of withdrawal bleeding compared to control animals. Of the five positive responses observed in the control females, four occurred when the higher dose of MPA was administered. CONCLUSION(S): These results indicate that the PCT is a useful, noninvasive method for determining the presence of endometrial stimulation in nonhuman primates.

Animals↗

Development of a potentially reversible vas deferens occlusion device and evaluation in primates.

A vas deferens occlusion device has been developed and tested in primates. The device consists of two silicone plugs held together by a nylon suture. Implantation of each plug is readily accomplished by making two small puncture holes in the vas deferens and inserting one plug into the lumen toward the epididymis and the other plug toward the seminal vesicles. Implantation is aided by a metal stylus inserted in each plug to give the plug rigidity. The stylus is removed after insertion of the plug into the vas deferens. Two separate primate experiments showed that the device completely prevents sperm transport over a 7-month period. Upon device removal, all of the primates ejaculated spermatozoa again at normal concentrations and motility. These results indicate the potential contraceptive use of the device and encourage its validation in men.

Humans↗

Nonhuman primate models of atherosclerosis: potential for the study of diabetes mellitus and hyperinsulinemia.

Nonhuman primates have been used for many years to investigate the pathogenesis and progression of atherosclerosis. The use of these animal models has resulted in a better understanding of the risk factors associated with atherosclerosis. Nonhuman primates that have consumed an atherogenic diet for several years develop lesions that are comparable to those found in human beings. Diabetes, both spontaneous and chemically induced, has been described in a number of nonhuman primate species. These diabetic models may be used to understand the accelerated progression and vascular complications of atherosclerosis in diabetic human beings.

Animals↗

Atherosclerosis and insulin in primates with diabetes mellitus.

The most commonly available primate models of diabetes mellitus are of the insulin-dependent type and are attained through beta cell ablation techniques. Noninsulin-dependent primate models are less common since the animals must have a genetic predisposition to diabetes. Few studies have been conducted on lipid or vascular abnormalities associated with diabetes in primates. Diabetes develops spontaneously in Macaca nigra as the result of a lesion in the islets of Langerhans. As secretory cells are gradually lost, mild to moderate hyperglycemia, impaired glucose clearance, acute insulin release, hyperglucagonemia, and chronic hypoinsulinemia develop. Overtly diabetic monkeys require insulin therapy and thus alternate between hypoinsulinemia and hyperinsulinemia. The development of aortic atherosclerosis correlates positively with the severity of metabolic impairment. Lipid deposition is primarily extracellular and there is a paucity of foam cells. The very low density and intermediate-density lipoprotein fractions increase significantly, the low-density lipoprotein fraction increases slightly, and the high-density lipoprotein fractions remain essentially unchanged. Because these monkeys are maintained on a nonatherogenic chow ration, the effects of diabetes, per se, on vascular sclerosis can be evaluated.

Animals↗

Formation and persistence of DNA adducts of 2-amino-3-methylimidazo[4,5-f]quinoline in the rat and nonhuman primates.

The formation and persistence of the two principal DNA adducts of the food derived carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) have been investigated in rats and nonhuman primates. DNA adduct formation in the liver of male Fischer-344 rats occurred in a dose-dependent manner (0.01-20 mg/kg) where N-(deoxyguanosin-8-yl)-2-amino-3-methylimidazo[4,5-f]quinoline (dG-C8-IQ) and 5-(deoxyguanosin-N2-yl)-2-amino-3-methylimidazo[4,5-f] quinoline (dG-N2-IQ) accounted for approximately 60-80% and 20-40%, respectively, of the total adducts observed by 32P postlabeling. Similar DNA adduct profiles were observed in kidney and colorectal tissue of rats given a single oral dose of IQ (20 mg/kg) which, when given chronically to this species, results in tumorigenesis in liver and colorectum, but not in kidney. dG-C8-IQ was removed more rapidly than dG-N2-IQ from liver and kidney, but removal of both adducts from the colorectum closely followed cell replication. Similar DNA adduct profiles were observed in liver and extrahepatic tissues of nonhuman primates following a single dose of IQ (20 mg/kg). In chronically treated monkeys undergoing carcinogen bioassay, there was a sharp increase in the contribution of dG-N2-IQ to total DNA adducts in all slowly dividing tissues. There was no preferential accumulation of dG-N2-IQ in the colon, a tissue with a high rate of cell division, and dG-C8-IQ remained the predominant lesion. These findings point to a preferential removal of the dG-C8-IQ adduct by enzyme repair system(s) in slowly dividing tissues in both rats and nonhuman primates.

Animals↗

Laterality of tail resting posture in three species of New World primates.

A variety of mammalian species including prosimian and simian primates wrap their tails around their bodies as a means of thermoregulation and for reasons of comfort during resting or sleep. Adopting such a resting posture requires an animal to move its tail either to the right or to the left of the midline of its body, and thus to perform a lateralized behavior. The purpose of this study was to assess the occurrence of lateral biases in tail resting posture in three species of New World primates. Twenty squirrel monkeys, spider monkeys, and howler monkeys, respectively, were observed and data on tail resting posture were collected and analyzed. The results demonstrate (1) that individual squirrel monkeys and spider monkeys exhibit highly significant lateral biases in tail resting posture; (2) a lack of a lateral bias at the group level; (3) that howler monkeys fail to show side preferences in tail wrapping; (4) a lack of sex differences in this behavior in all three species; and (5) a lack of significant correlations between preferred side of tail resting posture and preferred side of hand use in simple reaching tasks which had been assessed with a subset of animals in previous studies. Thus, the present study provides evidence for a behavioral asymmetry which is well-known to occur in rats but has not been described so far in nonhuman primates, and which might offer an additional approach to the investigation of the mechanisms underlying functional cerebral asymmetries.

Alouatta↗

Melatonin promotes sleep in three species of diurnal nonhuman primates.

Nocturnal melatonin secretion is concurrent with consolidated sleep episodes in diurnal mammals and physiological melatonin levels can promote sleep onset in humans and in pigtail macaques. In order to further investigate the effects of melatonin treatment on sleep parameters in diurnal nonhuman primates, three macaque species have been studied: Macaca nemestrina, Macaca fascicularis, and Macaca mulatta. Sleep was assessed using continuous actigraphic recording of motor activity in animals maintained under 12:12-h light/dark cycle. Oral doses of melatonin (5-320 microg/kg) were administered 2 h before lights-off time, with 5- and 10-microg/kg doses resulting in physiological circulating melatonin levels (31-95 pg/ml). The effects of melatonin administration were similar in three species studied and included significantly earlier sleep onset time and longer sleep period time, with no difference in time of awakening, following administration of both physiological (5-10 microg/kg) and pharmacological (20-320 microg/kg) doses. While low melatonin doses (5-20 microg/kg) did not significantly affect nighttime sleep efficiency, higher pharmacological doses reduced sleep efficiency and increased sleep fragmentation at night, and reduced spontaneous daytime locomotor activity. Daily administration of a 5-microg/kg dose for 4 weeks or gradually escalating melatonin doses (5-320 microg/kg over a 3-week period) did not result in the development of tolerance or sensitization to the effect of melatonin on sleep initiation or sleep period. These data affirm that sleep-promoting effects of melatonin observed in humans are also typical for diurnal primates. They also suggest that physiological and pharmacological melatonin levels might produce different effects on sleep efficiency and that nonhuman primates can serve as adequate animal model for studying the mechanisms of melatonin's action on sleep and performance.

Animals↗

Assessing olfactory performance in an Old World primate, Macaca nemestrina.

The present study demonstrates that an operant conditioning paradigm, originally designed for assessing olfactory performance in a small New World primate, the squirrel monkey, can successfully be adapted for use with a large Old World primate, the pigtail macaque. Using a task designed to simulate olfactory-guided foraging behavior, based on multiple discrimination of simultaneously presented odor stimuli, we could show that Macaca nemestrina is able to learn to discriminate between objects on the basis of odor cues. Moreover, they could readily transfer to new S+ and S- stimuli and could remember the significance of previously learned odor stimuli even after a 3-week break. Furthermore, we could show that this method is suitable for obtaining reliable measures of olfactory sensitivity. The few modifications of the original method employed here did not affect essential features such as the mode of stimulus presentation (odorized paper strips attached to manipulation objects) and the choice criterion (opening or rejecting the odorized manipulation objects), thus for the first time enabling valid interspecific comparisons of olfactory capabilities between a catarrhine and a platyrrhine primate species. Our results indicate that M. nemestrina and Saimiri sciureus are similar with regard to several measures of olfactory performance, such as speed of initial task acquisition and ability to master transfer tasks as well as their sensitivity to a food-related odorant.

Animals↗

Analysis of nonhuman primate peripheral blood mononuclear cells for susceptibility to HIV-1 infection and HIV coreceptor expression.

HIV-1 infection of nonhuman primates does not lead to the acquired immunodeficiency syndrome seen in humans. The basis for this lack of disease progression in these animals is still unknown. In this study, primary nonhuman primate peripheral blood mononuclear cells (PBMC) were tested for their susceptibility to in vitro infection by several different primary HIV-1 isolates representing distinct subtypes or clades. None of the five HIV-1 subtypes tested were able to readily establish an infection in chimpanzee or baboon PBMC, as determined by p24 antigen capture assays. To address the mechanism of in vitro resistance to HIV-1 infection, PBMC were analyzed for HIV coreceptor mRNA expression and cell surface expression. Flow cytometry analysis of the nonhuman primate PBMC demonstrated that they do express CD4, CCR3, CCR5, and CXCR4 on their cell surface. Therefore, the level of restriction in the virus replication cycle does not appear to lie at the point of entry in these cells.

Animals↗

Direct comparison of visual cortex activation in human and non-human primates using functional magnetic resonance imaging.

We report a technique for functional magnetic resonance imaging (fMRI) in an awake, co-operative, rhesus macaque (Macaca mulatta) in a conventional 1.5T clinical MR scanner, thus accomplishing the first direct comparison of activation in visual cortex between humans and non-human primates with fMRI. Activation was seen in multiple areas of striate and extra-striate visual cortex and in areas for motion, object and face recognition in the monkey and in homologous visual areas in a human volunteer. This article describes T1, T2 and T2* values for macaque cortex, suitable MR imaging sequences, a training schedule, stimulus delivery apparatus and restraining hardware for monkey fMRI using a conventional 19 cm knee coil. Much of our understanding of the functional organization of the primate brain comes from physiological studies in monkeys. Direct comparison between species using fMRI such as those described here will help us to relate the wealth of existing knowledge on the functional organization of the non-human primate brain to human fMRI.

Adolescent↗

Development of a competitive ELISA for detection of primates infected with monkey B virus (Herpesvirus simiae).

Two competitive ELISAs (C-ELISAs) are described that allow detection of antibodies against monkey B virus (BV, Cercopithecine herpesvirus 1). The assays utilize monoclonal antibodies (MABs) directed against the BV glycoprotein B (gB). Two of these MABs specifically recognize BV gB while a third MAB also reacts with the gB homologues of other primate alpha-herpesviruses (herpes simplexvirus-1, HSV-1: HSV-2; simian agent-8, SA8; and Herpesvirus papio-2, HVP2). A C-ELISA using the single cross-reactive MAB 3E8 allowed detection of host antibodies against HSV-1, HSV-2, SA8, HVP2 or BV, thus proving to be a sensitive assay for the detection of infection by any of these primate alpha-herpesviruses. The C-ELISA using BV-specific MABs was less sensitive but did allow some discrimination between infection by BV versus other alpha-herpesviruses. It was also shown that a C-ELISA using HVP2 as antigen and the cross-reactive MAB 3E8 was as sensitive for detection of BV antibody in macaque sera as an assay employing BV antigen. This test format allows detection of BV-infected primates without the biohazards associated with preparation and use of BV antigen.

Animals↗

The baboon as a non-human primate model of human schistosome infection.

Over the past three decades, intensive studies of murine schistosomiasis have provided important clues to the understanding of the human disease, but growing evidence suggests that these results derived from highly inbred strains of mice might not have direct applicability to the human infection. Recent data based on the baboon indicate that infection in this non-human primate might mirror the human situation. In this review, Mramba Nyindo and Idle Farah demonstrate that baboons provide an excellent non-human primate model that produces pathology and disease closely resembling that observed in humans, and address how studies in baboons can provide insights into mechanisms regulating schistosomiasis mansoni pathology and immunity. They also address, in a general way, issues related to the use of non-human primates in biomedical research.

Animal Welfare↗