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Effect of immunosuppression on morphological changes in CNS of monkeys infected with different measles virus vaccine strains.

Residual neurovirulence of measles virus vaccine strains "Leningrad-16", "Moscow-5" and of cloned variant of L-16 strain--clone 3, differing in the degree of attenuation, was studied in intracerebrally (i.c.) infected Cercopithecus aethiops monkeys, receiving cortisone and/or antilymphocyte serum (ALS). Whereas the sensitivity for detection of the potential neurovirulence of vaccine strains was not increased by cortisone treatment. ALS had such effect in detection of neurotropic properties of the L-16 strain and its cloned variants.

Animals↗

[Studies on the activity of the aryl hydrocarbon hydroxylase and metabolism of benzo(a)pyrene in cell strains and cell lines used for testing cytotoxic and transforming ability of foreign compounds (author's transl)].

The activity of the AHH and the formation of polar metabolites from BaP was examined in several cell strains and lines, which are used for testing cytotoxic and transforming activity of environmental agents, the highest activity of the AHH and the highest capacity to metabolize BaP was found in hamster kidney cells. Much lower values were measured in kidney cells from Cercopithecus aethiops, BHK21/C13 cells, and SV-40 transformed hamster kidney cells (14-1b). The activity of AHH was inducible by BA in all these cells. However no correlation could be established between the increase in AHH activity and the formation of polar metabolites. In cells derived from a human bronchial carcinoma (E-14), a very low BaP metabolism was detectable which increased 3-fold after exposure (o BA. No BaP metabolism could be detected in HeLa cells.

Animals↗

The use of murine monoclonal antibody B2M-01 for detection and purification of human beta 2-microglobulin.

A murine hybridoma cell line was obtained producing monoclonal antibody specific for human beta 2-microglobulin (beta 2m). The antibody reacts strongly with cell-associted human beta 2m, weakly with the cells of the monkey Cercopithecus aethiops, but not with any other non-primate cells tested. It is of IgG2a isotype, its pI is 6.9-7.1 and dissociation constant of its complex with soluble beta 2m is K = 1.5 X 10(-8) mol/l. The antibody also binds to beta 2m after electrophoretic separation under the reducing and denaturating conditions and blotting onto nitrocellulose. Immobilization of this antibody on Sepharose 4B yields an efficient immunosorbent for purification of beta 2m.

Antibodies, Monoclonal↗

An immunohistochemical study of the middle ear muscles of some carnivores and primates, with special reference to the IIM and slow-tonic fibre types.

The middle ear muscles of several species of carnivores (cat, dog, fox, ferret and stone-marten) and some New World monkeys (Callithrix, Saimiri) and Old World monkeys (Cercopithecus, Macaca) were examined. The fibre type compositions of these muscles were determined by a combination of the standard histochemical myofibrillar ATPase method, and immunohistochemical techniques using myosintype-specific antisera. Immunohistochemically slow-tonic fibres were found in the stapedius muscles of only two carnivores, the ferret and stone-marten. In all the carnivores and the New World monkeys, tensor tympani muscle contained IIM, slow-tonic and slow-twitch fibres, but in the Old World monkeys it resembled stapedius muscle, and contained only Type I (slow-twitch) and IIA fibres. Thus, because all the species examined had IIM fibres in the jaw-closer muscles, this means that the common embryological origin of tensor tympani muscle and the jaw-closers does not necessarily result in tensor tympani muscle containing this fibre type even though IIM fibres occur only in first branchial arch muscles. This fact, together with other species differences in the fibre type composition of these muscles, shows that there is no typical composition of middle ear muscles in general, and suggests that the differences are related to very different functional requirements.

Animals↗

Quantitative studies of the evolution of the thalamus in primates.

A quantitative study of the thalamus and metathalamus has been carried out on brains of six different primate species. The allometric formula devised by Stephan, Bauchot and Andy (1970) has been slightly modified as two steps have been utilized in order to attain the required magnification of the final photomicrograph (a positive print, as opposed to Stephan et al's negative print). The thalamic and metathalamic structures are divided on the basis of their developmental stages into two groups--paleothalamus and neothalamus, so that their proportions to each other in primate phylogeny can be estimated. The results show that the neothalamus extends the paleothalamus by two-thirds (2/3rds) in Tupaia and by more than three-quarters (3/4ths) in Cercopithecus. Of all the thalamic groups, the dorsolateral thalamic nuclear group is most dominant in all primate species, except Tupaia, where the ventrolateral thalamic nuclei are larger than all other nuclei. The pulvinar is the most dominant nucleus in all species except Tupaia. These quantitative results conform, in most places, with the observations of other researchers, while they do not agree much with the qualitative observations of the thalamus in the same species (Simmons 1974). How this study has a bearing on the phylogenetic positions of the Tupaioidea and the Tarsioidea in the Primate Order, is briefly discussed.

Animals↗

[Possible heterogeneity of cytoplasmic estrogen receptors].

In order to elucidate whether specific estrogen-binding sites were heterogeneous, these sites were studied in uterine cytosols of guinea pig and green monkey Cercopithecus griseus. Concentrations of the estrogen-binding sites for labelled estrogens (estrone, estradiol, estriol and diethylstilbestrol) were estimated in the cytosols by means of the exchange method and Scatchard plot analysis. High linear correlation was shown between the Scatchard plots and the curves of estrogen-binding site complexes dissociation plotted in semilogarithmic coordinates (r-0.98 and 0.99, respectively; p less than 0.05). The linearity of the curves was confirmed by means of F-criterion. The data obtained suggest that only one type of specific estrogen-binding sites occurs in the cytosols at 30 degrees and at concentration of estrogens which did not exceed 50 nM. These estrogen-binding sites exhibited the same limited binding capacity for all the estrogens studied.

Animals↗

[Local dermatitis followed by pigmentation in green vervet monkeys].

A group of 154 green vervets (Cercopithecus aethiops) was studied from a clinical and pathological point of view. In 27 male and two female animals a skin lesion was noticed on genitals. Physiological blue colour of these parts due to melanin cells in deep corium turned red at first; skin was smooth without itching and acute or subacute dermatitis was present in histology. Later smooth skin got dark or blackened because of epidermal melanin pigmentation though melanocytes are lacking in normal epidermis of these vervet parts. Etiology of skin lesions was not solved but pathological changes in melanocyte localisation were stimulating for the theory of melanocyte origin.

Animals↗

A surgical procedure and tethering system for chronic blood sampling, infusion, and temperature monitoring in caged nonhuman primates.

A jacket and tethering system was used to maintain chronic catheters in monkeys, which provided catheter access and manipulability without further restraint. Surgical placement of catheters and a temperature probe allowed for a common cutaneous exit and interface with the jacket and tether. Monkeys were fitted in a sterile leather or denim jacket which was attached to a sterilized flexible stainless steel cable. Through this conduit, an indwelling temperature probe, as well as catheters from the internal jugular and femoral veins, were attached to a swivel unit located on the upper portion of the cage. The internal jugular catheter was used for the continuous infusion of support solution. The catheter from the femoral vein was maintained with a heparin lock and used for serial blood sampling. Using this system, it was possible to obtain frequent blood samples and body temperature readings, and to administer a continuous intravenous infusion without chemical or excessive physical restraint. To date, 367 monkeys, 322 cynomolgus (Macaca fasicularis), 16 rhesus (Macaca mulatta), and 21 African green (Cercopithecus aethiops) have been studied using this procedure.

Animals↗

[Characteristics of ECHO-19 virus strain Rad/40 causing an experimental lesion of the vascular coat of the eye (uveitis) in cynomolgus monkeys].

Four strains of ECHO-19 virus were isolated from a child with respiratory disease complicated with uveitis. A freshly isolated Rad/40 strain of ECHO-19 virus was found to produce a severe disease of the vascular coat of the eye (uveitis) in Javan Cercopithecus monkeys inoculated directly into the anterior chamber of the eye. Experimental uveitis is characterized by marked changes in the shape of the pupil, the lack of the pupil reaction to light, development of synechias, changes in the color of the iris. No uveitis developed after inoculation of the monkeys with the prototype Burke strain of ECHO-19 virus of culture fluid from uninoculated RD cell cultures (control). It is suggested that the freshly isolated strains of ECHO-19 virus (Rad/40 strain) could cause uveitis in the child under observation.

Animals↗

Cholesterol absorption and transport in thoracic duct lymph lipoproteins of nonhuman primates. Effect of dietary cholesterol level.

The effect of dietary cholesterol level on cholesterol absorption and its subsequent transport in thoracic duct lymph lipoproteins was studied in two species of nonhuman primates, namely the African green monkey (Cercopithecus aethiops) and the cynomolgus macaque (Macaca fascicularis). Each animal served as its own control because each received sequential, intraduodenal infusions of two fat-rich liquid diets that differed only in the amounts of cholesterol. The percentage of dietary cholesterol absorbed was calculated by dividing the rate of appearance of exogenous cholesterol radioactivity in lymph by the rate of infusion of dietary cholesterol radioactivity at a time when lymph cholesterol specific activity was constant relative to that of diet. The percentage of dietary cholesterol absorbed was similar during both diet infusions in African green monkeys but was significantly decreased during the high cholesterol diet infusion in cynomolgus macaques. Rates of appearance of lymph total cholesterol mass were significantly increased during infusion of high cholesterol diets due to a statistically significant increase in cholesteryl ester transport rates. This increase was due in large part to the preferential esterification of exogenous cholesterol that was incorporated into lymph chylomicrons and VLDL. The rate of appearance in lymph of exogenous cholesterol significantly increased during the high cholesterol diet infusion while that of endogenous cholesterol decreased. This decrease or compensation in endogenous cholesterol transport occurred during absorption of increased levels of dietary cholesterol and apparently was due to an approximate 50% decrease in the absorption of lumenal cholesterol. Our data demonstrated that there was not a simple one-to-one relationship between the amount of isotopic dietary cholesterol absorbed from the intestinal lumen and the mass of cholesterol moved into lymph via the intestine. Rather, the amount of sterol transported into the body depends on the degree of the decrease in endogenous cholesterol transport and probably on the efficiency of cholesterol esterification during absorption.

Animals↗

Analysis of pathogenesis and topography of replacement root resorption (ankylosis) after replantation of mature permanent incisors in monkeys.

The pathogenesis and topography of replacement root resorption (ankylosis) after replantation of incisors was examined in green Vervet monkeys (Cercopithecus aethiops). Incisors were replanted after 0,18 or 120 min. storage and examined histometrically for replacement resorption after 2, 4 and 8 weeks. There was a significant preference for certain locations upon the root surface. Thus, ankylosis was found significantly more often on the rounded labial and lingual surfaces than on the flat or concave proximal surfaces, and on the apical portion of the root in contrast to the coronal portion. At two-weeks' observation periods, two types of initial ankylosis were seen after 120 min. dry storage. In the cervical areas and on the labial and lingual surfaces, the most common finding was a complete mineralization of the entire periodontal ligament, while on the remaining parts of the root surface, the initial ankylosis area consisted of a layer of bone deposited upon the root surface and socket wall with an intervening soft connective tissue zone. These two zones were in some areas united by bony bridges formed along the Sharpey's fiber bundles. It is suggested that the first type of initial ankylosis represents areas where both the cemental and alveolar parts of the periodontal ligament are avital; whereas the second type represents areas where only the cemental part of the periodontal ligament is avital (i.e., from dry storage). It is concluded that ankylosis after replantation is presumably a response in the periodontium to areas of the periodontal ligament and/or the root surface damaged by the extraction procedure or storage conditions prior to replantation.

Animals↗

Morphological evidence that hypothalamic substance P-containing afferents are capable of filtering the signal flow in the monkey hippocampal formation.

This study in the African green monkey (Cercopithecus aethiops) was designed to characterize the neurochemical features of hippocampal nonpyramidal neurons that are specific synaptic targets of substance P-containing projective neurons located in the supramammillary nucleus. Our previous studies provided evidence for an excitatory nature to this hypothalamo-hippocampal pathway and described the mode of termination of these afferents on hippocampal principal neurons. The present correlated light and electron microscopic immunocytochemical analysis, using the nickel-diaminobenzidine/diaminobenzidine double-labeling technique, revealed that this hippocampal afferent system establishes multiple, exclusively asymmetric synapses with three specific subpopulations of nonpyramidal cells: (1) a small portion of parvalbumin-containing basket cells located periodically in or adjacent to the granule cell layer of the dentate gyrus, which therefore inhibit only a subpopulation of granule cells; (2) some of the calbindin-immunoreactive local circuit neurons located in the hilar area; and (3) calbindin-positive cells occurring exclusively in the stratum molecular of the middle portion of the CA3 subfield. Postembedding studies revealed that the aforementioned calbindin-containing cells are GABAergic inhibitory neurons. Our studies indicate that hypothalamic afferents can effectively filter the information flow at different levels of the excitatory signal loop in the monkey hippocampal formation. Dentate granule cells, which are only stimulated by hypothalamic afferents, will transfer excitatory signals differently than those that are controlled by a feedforward inhibitory mechanism initiated by these fibers. In the CA3 subfield, the signal flow can again be depressed by those pyramidal neurons that are inhibited by calbindin-containing cells receiving an excitatory hypothalamic input.

Animals↗

[Percutaneous absorption of phenanthrene: an in vitro study of the monkey skin].

It is well known that polycyclic aromatic hydrocarbons (PAHs) are absorbed through the skin of experimental animals and humans. However dermal uptake of PAHs is difficult to assess due to the lack of specific studies. The aim of this study was to obtain in vitro percutaneous absorption data for phenanthrene. In vitro penetration was measured with excised monkey (Cercopithecus Aethiops) skin in Franz diffusion cells, using a saline solution with 4% bovine serum albumin and gentamicin sulfate as receptor fluid. In two different groups of 7 and 2 cells, 38.1 and 95.3 nmol/cm2 respectively of phenanthrene were applied in an acetone vehicle. The absorption rate in the first group of 7 cells was 0.025 nmol/cm2/h (S.D. = 0.012), the lag time 11.7 (S.D. = 7.0) h and the breakthrough time 70 m (S.D. = 55 m). In the second group of 2 cells, the average absorption rate was 0.066 nmol/cm2/h and the average lag time 11.8 h. The percutaneous absorption at 24, 48 and 72 hours in the 7 cell group was 0.72% (S.D. = 0.52), 2.51% (S.D. = 1.76) and 3.9% (S.D. = 2.0) of the applied dose respectively. In the 2-cell group 1.1%, 2.78% and 4.5% of the applied dose was absorbed at 24, 48 and 72 hours. The results of the present study confirm the extent of percutaneous absorption of phenanthrene. The data can be compared with dermal absorption values of other PAHs obtained under the same experimental conditions.

Animals↗

Distribution of calbindin D28k immunoreactive cells and fibers in the monkey hippocampus, subicular complex and entorhinal cortex. A light and electron microscopic study.

Calbindin D28k (CB)-containing neurons and axon terminals in the hippocampus, subicular complex and entorhinal cortex of the African green monkey (Cercopithecus aethiops) were studied by light and electron microscopic immunocytochemistry. CB was present in granule cells of the dentate gyrus, pyramidal neurons of hippocampal fields CA1 and CA2, and in pyramidal neurons of the prosubiculum and entorhinal cortex. In contrast, pyramidal neurons in the CA3, subiculum and presubiculum were not labeled. A subpopulation of non-principal neurons (non-granule and non-pyramidal cells) was also stained for CB. These cells were rare in the hippocampus and subiculum, but were more frequently observed in the presubiculum, parasubiculum, and in the entorhinal cortex. In the electron microscope, these non-principal cells displayed fine-structural characteristics of GABAergig neurons. Strongly stained CB-immunoreactive bundles of myelinated axons were found in the molecular layer of the subiculum and in various layers of the presubiculum. The CB-positive, unmyelinated axons of the granule cells, the mossy fibers, gave rise to distinct fiber bundles. Mossy fiber terminals formed asymmetric synapses on large spines in the hilus and CA3. In addition to the giant mossy fiber boutons, there were large CB-positive terminals that formed asymmetric synapses with dentritic spines throughout the hippocampal formation. These boutons also formed axo-dendritic synapses in the entorhinal cortex. Axon terminals that formed symmetric synapses and might, thus, be derived from non-principal neurons, were rarely found in the hippocampus and subicular complex. They were more frequent in the parasubiculum and entorhinal cortex. These CB-positive terminals were small, heavily immunostained, and formed symmetric axo-dendritic synapses. Our results demonstrate a great diversity of CB-containing neurons, axons, and terminals in the monkey hippocampal formation. In general, regions that received a dense innervation of CB-positive terminals displayed pyramidal neurons that all lacked this calcium-binding protein. Further studies are required to understand the functional significance of these findings.

Animals↗

A randomized, placebo-controlled, blind anti-AIDS clinical trial: safety and immunogenicity of a specific anti-IFN alpha immunization.

HIV-induced cytokine dysregulation, including overproduction of the antiproliferative and cytolytic IFN alpha cytokine, represents a major component of the immune disorders characterizing AIDS. To block the overproduction of IFN alpha we designed an AIDS vaccine combination which included both an anti-HIV and/or an anti-IFN alpha immunization. The safety and immunogenicity of this multicomponent vaccine were tested in mice, Cercopithecus, two HIV noninfected individuals, and six HIV-1 seropositive immunocompromised patients enrolled in a 1-year open clinical trial. We now report the result of a 9-month short-term randomized, blind, placebo-controlled clinical trial (Phase I/II) performed in HIV-1 patients (22 individuals) to confirm safety/tolerance of the anti-IFN alpha vaccine and its immunogenicity and to evaluate whether the complex vaccine initially used could be simplified by removal of HIV component(s). Three groups of patients received inactivated IFN alpha (i-IFN alpha) associated with the immunomodulator P40 with HIV-1 antigens (groups B and C) or without (group A), and one group (D) was placebo. The clinical follow-up documented among those receiving i-IFN-alpha showed that none developed AIDS and/or required antiretroviral chemotherapy. Viral load did not increase and CD4 cell count as well as cell-mediated immunity (CMI) stabilized or even significantly increased in group A. Immunogenicity of the preparations was determined by a positive delayed-type hypersensitivity (DTH) reaction to i-IFN alpha and the presence of serum antibodies to i-IFN alpha and to HIV-1 peptides, occurring only in treated patients. As previously planned, based on these safety data, the trial has been extended for an additional year and all patients were switched to protocol A (i-IFN alpha+P40). This second period of the trial, now open and ongoing, should allow us to evaluate further the innocuity of the i-IFN alpha preparation and whether anti-IFN alpha vaccine could provide a long-lasting CD4 cell count as well as CMI stabilization.

AIDS Vaccines↗

Tunica vaginalis sperm reservoir in a monkey model of vas deferens obstruction.

Repeat microscopic epididymal sperm aspiration (MESA) is used to treat congenital absence of the vas deferens (CAVD). Use of alloplastic implants as reservoirs results in poor sperm motility and pregnancy rates. An autoplastic tunica vaginalis reservoir for epididymal sperm has been used in four patients, with one resultant pregnancy. We studied this technique in a monkey model. Eight reservoirs were created in four monkeys (Cercopithecus aethiops). The vas deferens was ligated close to the epididymis. A transverse incision was made in the caput epididymis, and sperm were aspirated for analysis. Cut margins of the epididymal duct were sutured open to the visceral layer of the tunica vaginalis. The defect in the parietal layer of the tunica vaginalis was repaired, and sterile Tyrode's solution was instilled in the potential space between the parietal and visceral layers. Initial epididymal sperm obtained at the creation of the sperm reservoir demonstrated adequate mean motility (60.5%, grade 1.5), morphology (82.8% normal), and viability by Eosin-Y exclusion (69.9%). At 4 weeks, no reservoirs were palpable. Percutaneous aspiration (25-gauge angiocatheter) yielded no fluid. Surgical exposure of the reservoirs demonstrated significant adhesions and scar formation between the parietal and visceral layers of the tunica vaginalis, obliterating the potential space. No sperm were detected in irrigant fluids at this time. In conclusion, adhesion and scar formation may prevent use of the tunica vaginalis reservoir as an alternative to repeat MESA in treatment of CAVD.

Animals↗

[The isolation and characteristics of the green monkey lentivirus].

We characterized the simian immunodeficiency virus isolated from Cercopithecus aethiops (subspecies C. a. pygerythrus) originating from Kenya. SIV was isolated and continuously produced with the MOLT4 clone 8 cell line and was designated as SIV-SU1. SIV-SU1 isolate replicated with high efficiency in MOLT4 clone 8, MT-2 with moderate efficiency in CEM x 174 and with poor efficiency in HUT-78, U937, C8166. The infection of MT-2, C8166 and HUT-78 resulted in extensive cell killing. Western blotting of purified preparations of SIV-SU1 revealed viral proteins of 130, 68, 55, 41, 24, 17 kDa. Cross-reactivity of SIV-SU1 proteins with HIV-1, HIV-2, SIVmac, SIVsm, SIVmnd was studied by radioimmunoprecipitation assay. The most extensive cross-reactivity was observed with SIVmac. Total cellular DNA from chronically infected cells was hybridized to SIVagm266 DNA probes. Detection of cross-hybridizing DNA sequences required very low stringency, and the restriction endonuclease fragmentation pattern of SIV-SU1 differed from other SIVs.

Animals↗

[Experimental models of hepatitis A in macaques using viral strains isolated from man and monkeys].

Experimental hepatitis A (HA) models were obtained in macaca monkeys (15 M. fascicularis and 4 M. mulatta) by means of the strains of hepatitis A virus (HAV) isolated from the feces of a patient (HAV-H) and of spontaneously infected M. Mulatta (HAV-MM) and green monkeys Cercopithecus aethiops (HAV-CA). Irrespective of the strains used all seronegative macaca monkeys developed HA after intravenous-oral inoculation with the following patterns: elevation of the serum alanine aminotransferase level, HAV shedding in feces, seroconversion with the appearance of anti-HAV IgM and morphological changes in the liver characteristic of acute hepatitis. HAV in fecal samples and elevation of alanine aminotransferase were periodically detected. Periods of their discovery varied from 5-22 to 15-47 days and those of morphological changes in the liver from 9-24 to 40-83 days. The results of the experiments show that experimental HA models in Macaca monkeys are no less adequate than the previous ones developed in chimpanzees (Pan troglodytes), marmosets (Saguinus mystax) and owl monkeys (Aotus trivirgatus), but they are more readily available. Both strain HAV-H and strains from monkeys can be used for HA modelling. The models are expected to be used for studying yet unsolved problems of pathogenesis and immunogenesis, as well as for testing vaccines and antiviral drugs.

Animals↗