African Green Research Articles

Cell attachment and spreading on extracellular matrix-coated beads

Parietal yolk sac cells M1536-B3 grown on cytodex 2 beads deposited an extracellular matrix on the surface of the beads. Cell-free matrix-coated beads were isolated by treatment of the cell monolayer with cytochalasin B (CB) at a concentration of 10 μg/ml of phosphate-buffered saline (PBS). The matrix when analysed by electrophoresis on polyacrylamide gels (PAGE) revealed that the major components were laminin and entactin. The matrix-coated beads were used to study the attachment, spreading, and growth of African Green monkey BSC-40, human mammary MCF-7, mouse fibroblast L929, rat liver clone 9, and rat hepatoma H-4-II-E cells in defined serum-free growth medium. The different cell lines exhibited varying responses to matrix-coated vs uncoated beads with respect to rate of attachment, spreading, and growth. One of the most consistent responses observed was the enhancement of cell spreading on matrix-coated beads. The results suggested that the matrix-coated beads will provide a readily available and valuable tool for studies on cell surface-extracellular matrix interactions and the physiological consequences of those interactions.

Structure of extrachromosomal circular DNAs containing both the Alu family of dispersed repetitive sequences and other regions of chromosomal DNA

Small polydisperse circular DNA (spcDNA) isolated from the BSC-1 line of African Green monkey kidney cells was digested with the restriction endonuclease BamHI and cloned in bacteriophage lambda. The resulting library of 25,000 phage was then screened for the presence of the Alu family of short interspersed nucleotide sequences, and four of the 100 Alu-positive clones were characterized. In summary: (1) all four clones contained regions other than Alu that were homologous to the BSC-1 chromosome. Two contained Alu plus unique chromosomal DNA, one contained Alu plus an uncharacterized repetitive chromosomal DNA, and one contained Alu plus both unique and a specific tandemly repeated chromosomal DNA (α-satellite); (2) all four clones were derived from extrachromosomal circular DNAs and not from the accidental cloning of a very small amount of contaminating chromosomal material assumed to be present in spcDNA preparations; and (3) one clone represented an intact circular DNA with a restriction endonuclease cleavage map that was a circularly permuted version of its chromosomal homologue.

Some extrachromosomal circular DNAs containing the Alu family of dispersed repetitive sequences may be reverse transcripts

A 300 base-pair (bp) size class of small polydisperse circular DNA (spcDNA) isolated from the BSC-1 line of African Green monkey kidney cells was cleaved with the restriction endonuclease Sau3A, and the resulting fragments (100 to 200 bp) were cloned in bacteriophage M13 mp7. The nucleotide sequence of each of 24 clones containing DNA sequences homologous to the Alu family of mobile, dispersed, repetitive elements was then determined. Analysis of these sequences revealed that many, and perhaps all, of the 300 bp Alu-containing spcDNAs had regions in which the 5′ and 3′ ends of the normal Alu element were juxtaposed and covalently joined. Although more than one model can explain the generation of such circular molecules, the most attractive one at this time involves their generation from reverse transcripts of Alu-specific RNAs.

Effects of three methods of restraint an in rhesus and african green monkeys intravenous glucose tolerance testing.

The intravenous administration of 0.75 gm glucose per kg and the measurement of serum glucose pretest and at 10, 20, 30, 60, 90 and 120 minutes constitute a satisfactory protocol for intravenous glucose tolerance testing of Rhesus (Macaca mulatto) and African Green (Cercopithecus aethiops) monkeys. No significant differences were noted between animals restrained with ketamine hydrochloride and those restrained with sodium pentobarbital, but the African Green males and females and the male Rhesus monkeys yielded significantly different results while being manually restrained.

Influenza virus hemagglutinin expression is polarized in cells infected with recombinant SV40 viruses carrying cloned hemagglutinin DNA

Primary cell cultures of African Green monkey kidney (AGMK) contain polarized epithelial cells in which influenza virus matures predominantly at the apical surfaces above tight junctions. Influenza virus glycoproteins were found to be localized at the same membrane domain from which the virus budded. When polarized primary AGMK cells were infected with recombinant SV40 viruses containing DNA coding for either an influenza virus H1 or H2 subtype hemagglutinin (HA), the HA proteins were preferentially expressed at the apical surface in a manner identical to that observed in influenza virus-infected cells. Thus, cellular mechanisms for sorting membrane glycoproteins recognize some structural feature of the HA glycoprotein itself, and other viral proteins are not necessary for this process.

Subchronic Toxicity of Chlorine Dioxide and Related Compounds in Drinking Water in the Nonhuman Primate

Subchronic toxicities of ClO2, NaClO2, NaClO3 and NH2Cl were studied in the African Green monkeys (Cercopithecus aethiops). The chemicals were administered in drinking water during 30-60 days subchronic rising dose protocols. The only unexpected and significant toxic effect was elicited by ClO2; this chemical inhibited thyroid metabolism in the animals at a dose of ca. 9.0 mg/kg/day. A statistically significant decrease of serum thyroxine occurred after the fourth week of exposure to 100 mg/l.concentration. The extent of thyroid suppression was dose dependent in each individual monkey, and was reversible after cessation of exposure. NaClO2 and NaClO3 failed to elicit similar effects in doses up to ca. 60 mg/kg/day. Also, NaClO4 or NH2Cl did not cause T-4 suppression in doses of 10 mg/kg/day. The selective thyroid effect of ClO2 was unexplained and it appeared to be paradoxical since ClO2 was rapidly reduced by the oral and gastric secretions to nonoxidizing species (presumably Cl-). No evidence of thyroid effects were detected in the serum of human volunteers who ingested approximately 1 mg/l. of ClO2 in drinking water as a result of routine use in the community water treatment process. Sodium chlorite induced dose-dependent oxidative stress on hematopoesis, causing decreased hemoglobin and red cell count and increased methemoglobin content. At the same time, serum transaminase (SGPT) levels showed significant subclinical elevation. The hematologic effects of NaClO2 rebounded during exposure indicating compensatory hemopoietic activity taking effect during oxidative stress. Sodium chlorate and chloramine did not induce detectable hematologic changes in the animals.

Subchronic toxicity of chlorine dioxide and related compounds in drinking water in the nonhuman primate.

Subchronic toxicities of ClO2, NaClO2, NaClO3 and NH2Cl were studied in the African Green monkeys (Cercopithecus aethiops). The chemicals were administered in drinking water during 30-60 days subchronic rising dose protocols. The only unexpected and significant toxic effect was elicited by ClO2; this chemical inhibited thyroid metabolism in the animals at a dose of ca. 9.0 mg/kg/day. A statistically significant decrease of serum thyroxine occurred after the fourth week of exposure to 100 mg/l.concentration. The extent of thyroid suppression was dose dependent in each individual monkey, and was reversible after cessation of exposure. NaClO2 and NaClO3 failed to elicit similar effects in doses up to ca. 60 mg/kg/day. Also, NaClO4 or NH2Cl did not cause T-4 suppression in doses of 10 mg/kg/day. The selective thyroid effect of ClO2 was unexplained and it appeared to be paradoxical since ClO2 was rapidly reduced by the oral and gastric secretions to nonoxidizing species (presumably Cl-). No evidence of thyroid effects were detected in the serum of human volunteers who ingested approximately 1 mg/l. of ClO2 in drinking water as a result of routine use in the community water treatment process. Sodium chlorite induced dose-dependent oxidative stress on hematopoesis, causing decreased hemoglobin and red cell count and increased methemoglobin content. At the same time, serum transaminase (SGPT) levels showed significant subclinical elevation. The hematologic effects of NaClO2 rebounded during exposure indicating compensatory hemopoietic activity taking effect during oxidative stress. Sodium chlorate and chloramine did not induce detectable hematologic changes in the animals.

Toxic Potential of the Plasticizer Di(2-ethylhexyl) Phthalate in the Context of Its Disposition and Metabolism in Primates and Man

Although human toxicity from exposure to the plasticizer di(2-ethylhexyl) phthalate (DEHP) is unknown, reports of animal toxicity from DEHP have stimulated extensive toxicological studies. In the absence of direct toxicity data, information on the disposition and metabolism of DEHP in primates and man may enhance our assessment of the toxic potential of DEHP in man. Studies of DEHP disposition and metabolism in the African Green monkey and man show that the compound is rapidly and extensively metabolized. It is excreted largely in the urine (greater than 90%) as conjugated (glucuronide) oxidation products of mono(2-ethylhexyl) phthalate; excretion in feces accounts for the other 10% of the administered DEHP. Plasma disappearance of parenterally administered DEHP is equally rapid so that by 24 hr following DEHP administration, plasma DEHP concentrations are virtually undetectable, while greater than 70% of the dose has been excreted in urine and stool. The transience of DEHP in primates and the extent to which it is metabolized and conjugated may play a role in the observed lack of toxicity.

Toxic potential of the plasticizer Di(2-ethylhexyl) phthalate in the context of its disposition and metabolism in primates and man.

Although human toxicity from exposure to the plasticizer di(2-ethylhexyl) phthalate (DEHP) is unknown, reports of animal toxicity from DEHP have stimulated extensive toxicological studies. In the absence of direct toxicity data, information on the disposition and metabolism of DEHP in primates and man may enhance our assessment of the toxic potential of DEHP in man. Studies of DEHP disposition and metabolism in the African Green monkey and man show that the compound is rapidly and extensively metabolized. It is excreted largely in the urine (greater than 90%) as conjugated (glucuronide) oxidation products of mono(2-ethylhexyl) phthalate; excretion in feces accounts for the other 10% of the administered DEHP. Plasma disappearance of parenterally administered DEHP is equally rapid so that by 24 hr following DEHP administration, plasma DEHP concentrations are virtually undetectable, while greater than 70% of the dose has been excreted in urine and stool. The transience of DEHP in primates and the extent to which it is metabolized and conjugated may play a role in the observed lack of toxicity.

An evaluation of dipsogenic stimuli in the African green monkey.

Elevations in the concentration of plasma angiotensin II (AII) and decline in plasma aldosterone (Ald) were noted in African Green monkeys at 48 hr of water deprivation but not subsequent to an equivalent duration of food deprivation, compared with nondeprived levels. In a second experiment, drinking was initiated following treatment with AII, hypertonic saline, and the beta-adrenergic stimulator isoproterenol. Concomitant elevations in plasma AII concentrations were measured following isoproterenol injection, but not after AII or hypertonic saline injection, when compared with isotonic saline treatment. Elevations in plasma Ald levels were noted following AII injection. A third experiment evaluated dipsogenic additivity of stimuli by comparing the volumes of water consumed following isoproterenol or hypertonic saline injection with the intake resulting from combined treatment with isoproterenol and hypertonic saline. Additivity was tested under ad lib conditions and following adaptation to a daily water deprivation regimen. The results of the first two experiments generally agree with predictions based on the respective contributions by intracellular dehydration and extracellular fluid volume depletion, to thirst. However, additivity of thirst stimuli was not demonstrated in the third experiment.

The integrated SV40 genome in permissive transformed monkey cells

The organization of the viral DNA sequences in three clonal lines of SV40-transformed permissive African Green monkey kidney cells was investigated by digesting the cellular DNA with restriction endonucleases and hybridizing the specific DNA fragments with labeled SV40 DNA. The genome of each transformed line contained a single insert of viral sequences located at a position which differed from line to line. No evidence for extrachromosomal SV40 DNA was obtained. Restriction mapping of the inserted SV40 DNA indicated that integration occurred without gross tandem duplication and that an intact early region was retained. Alterations were observed in the late region of the SV40 inserts. Although these alterations were not identical in the three transformed clonal populations, the HaeII restriction endonuclease recognition site at 0.83 SV40 map unit was absent in two of the three lines.

Sequence relationships between single repeat units of highly reiterated African Green monkey DNA

Individual monomer and dimer units of the highly repeated alpha-component DNA of African Green monkeys were isolated and amplified by molecular cloning in pBR322. The purified sequences were characterized by digestion with restriction endonucleases and by primary nucleotide sequence analysis. Comparison of the cloned units with the 172 base pair long sequence representing the most abundant nucleotide at each position in the set of sequences comprising alpha-component allows the following conclusions. The set of sequences comprising alpha-component is made up of a very large number of related but slightly divergent sequences. Two neighboring repeats of the monomer unit are not necessarily more similar to one another than are randomly isolated monomers.

Hemagglutination typing of Escherichia coli: definition of seven hemagglutination types.

A hemagglutination (HA) typing system has been developed for demonstrating and characterizing the mannose-sensitive and mannose-resistant hemagglutinins produced by Escherichia coli isolated from human sources. HA typing is performed by testing CFA agar-grown E. coli cells for HA with human, bovine, adult chicken, African Green monkey, and guinea pig erythrocytes in the presence and absence of mannose. Seven major HA types, designated HA type I through HA type VII, have been defined according to the HA patterns produced by 1,334 test cultures consisting of 33 colonization factor antigen I (CFA/I)-positive enterotoxigenic E. coli (ETEC), 37 CFA/II-positive ETEC, 614 isolates belonging to the classical enteropathogenic E. coli, or EPEC, serogroups, 446 non-ETEC, non-EPEC stool isolates, and 204 bacteremia-associated E. coli. Facultatively enteropathogenic E. coli (FEEC) serogroups, which are the causative agents of extraintestinal infections but also sporadic cases of enteritis, comprised 38% of the stool isolates and 91% of the blood isolates examined. Previous observations concerning the HA patterns of CFA-positive ETEC and the EPEC were confirmed. A significant correlation was found between FEEC serogroups and the production of mannose-resistant HA with human, monkey, and usually chicken erythrocytes (the HA patterns designated HA type VI). A large majority (80.2%) of the FEEC strains belonging to the most frequently isolated serogroups from cases of bacteremia (O1, O2, O4, O6, O7, and O18) produced type VI HA patterns. Stool isolates belonging to these same serogroups were 59.2% positive for HA type VI patterns. In contrast, only 17.4% of the non-FEEC stool isolates and 1.9% of the EPEC isolates belong to HA type VI. Of the blood isolates, the HA type VI phenotype was two times more prevalent among K1-positive E. coli than among K1-negative E. coli, 70.6 versus 31.1%. These results suggest that surface-associated hemagglutinins of E. coli, many of which are known to be fimbriae, should be considered in addition to serotype (O:K:H antigenicity) in the description of isolates.

Experimentally-Induced Symptoms and Host Range of Citrus Likubin (Greening Disease)

In an attempt to find suitable indicator plants for citrus likubin (greening disease occurring in Taiwan) a series of experimental tissue-inoculation tests was made on plants of various citrus species and hybrids and other rutaceous plants grown and maintained in the greenhouse. Typical leaf symptoms in citrus were various chlorotic patterns including some resembling zinc deficiency, also blotchy mottle, yellowing, shortening of internodes and stunting (Fig. 1, A-D). In the inoculated seedlings and budlings irregular chlorotic mottle along the veins was produced in 2 or 3 months at 28-32 C, the most favorable temperature conditions for symptom development. At the earlier stage of infection the symptoms tended to appear irregularly in parts of the inoculated plants, then gradually progressed throughout plants, forming shortened internodes, small leaves and sometimes vein corking (Fig. 1, E, F and B). In this series of inoculation tests, however, consistently high rates of infection were obtained by inoculation with 2 or 3 inoculum buds in each seedling or budling (Table 1). Among the citrus varieties tested, sweet orange (Citrus sinensis), mandarins (C. reticulata) and Orlando tangelo (C. paradisi×C. reticulata) were found to be highly responsive to the likubin agent and likubin symptoms were clearly defined despite the presence of seedling yellows tristeza virus which is widespread in mandarin citrus trees in Asian countries. In contrast, Eureka lemon (C. limon), sour orange (C. aurantium), grapefruit (C. paradisi), and Sexton tangelo seedlings were much more sensitive to seedling yellows tristeza virus than to the likubin pathogen. The blotchy mottle which developed on leaves of inoculated rough lemon (C. jambhiri) was very similar to that of South African greening disease. Ponkan (C. reticulata) and Orlando tangelo seedlings showed especially conspicuous leaf symptoms in these tests (Fig. 1, A and Fig. 3). Therefore, it is suggested that these varieties be recommended as indicator plants for likubin, even if the inoculum carries seedling yellows tristeza virus. Kumquat varieties (Fortunella spp.) also developed chlorotic and yellowing leaf symptoms typical of likubin. No distinct symptom appeared on trifoliate orange (Poncirus trifoliata) seedlings; however, sub-inoculated seedlings were found to be infected by topgrafting them with susceptible citrus varieties. Murraya (Murraya paniculata), a favorite host plant for Diaphorina citri, the psyllid vector of greening disease, did not show any visible sign of infection following tissue-inoculation.

The results of geographic isolation on the teeth and skull of the Green monkey (Cercopithecus aethiops sabaeus) in St. Kitts‐a multivariate retrospect

A population of the West African Green monkey (Cercopithecus aethiops sabaeus) has been established for some 300 years (100 generations) on the West Indian island of St Kitts. Earlier studies carried out between 20 and 30 years ago showed this island population to have greater dental and cranial dimensions, to be less variable in these dimensions, to be more variable in certain meristic dental characters (e.g. supernumerary teeth) and to be less symmetrical in dental and cranial features than the contemporary mainland descendants of the parent West African stock. The extent of divergence in dental and cranial dimensional characters between the West Indian and West African Green monkeys appeared to be on the same general scale as that obtaining between the West African Green monkey and certain other groups of African cercopitheques commonly accorded distinct subspecific or specific status.These earlier analyses were unavoidably based upon univariate statistical techniques or upon simple derivatives therefrom. Hence, it was not then possible to evaluate correlations between dimensions so that detailed results may conceivably have been influenced by this and by other inescapable factors, e.g. lack of mensural data occasioned by sample limitation and the imperfection of particular specimens. It is also possible, if unlikely, that these earlier results might have been influenced by a systematic computational error, resulting in the variance of bilateral dimensions being increased by a factor of approximately two.The original data (slightly augmented by newly available material) have been reanalysed by univariate and multivariate techniques designed to overcome former deficiencies, the latter now being deployable because of the availability of computers of adequate power.Such new analyses have fully substantiated the earlier findings and have given precision to the comparison of the extent of divergence in a total of 73 dental and cranial dimensions between the St. Kitts Green monkey, the African Green monkey and other species and subspecies of Cercopithecus.The overall divergence in the sum total of dental and cranial dimensions as measured by the generalized distance (a multivariate quantity compounding both size and shape) between three subspecies (sabaeus, pygerythrus and aethiops) of Cercopithecus aethiops has proved to be of the same general scale as that between males and females in Cercopithecus and to be smaller than that between three full species (aethiops, nictitans and cephus) of this genus.Divergence between the St. Kitts and African Green monkeys in the combination of features has emerged as being generally similar to that between species of the genus Cercopithecus, and bigger than that between subspecies.Analysis of cranial dimensions alone has given a similar pattern of separation. In contrast, a multivariate compound of dental dimensions has revealed the extent of divergence between subspecies, between sexes as well as between species as being similar, the divergence between the St. Kitts and African population being also of the same general order.The pattern of dental and cranial divergence between the St. Kitts Green monkey, the African Green monkey and other cercopitheques is similar if factors relating to size are eliminated from the analysis and attention confined to shape only, irrespective of whether or not such components attributable to proportional growth changes (size‐dependent shape), or to those which are apparently independent of overall size, are included in the analyses.Such differences could well betoken a measure of genetic contrast between the present‐day St. Kitts and West African Green monkey but the new information elicited in the present study, even when combined with the results of recent studies of the behaviour and sociology of the St. Kitts Green monkey, and of certain aspects of its reproductive biology, does not necessarily form a basis for according the island monkey a taxonomic status distinct from that of the present‐day African descendants of its parent stock: Cercopithecus aethiops sabaeus.

Evidence for the existence of a single enzyme catalyzing the phosphorylation of choline and ethanolamine in primate lung

Choline kinase (ATP: choline phosphotransferase, EC 2.7.1.32) has been isolated and purified 1000-fold from adult African Green monkey lung with a yield of 10%. The purified enzyme also phosphorylated ethanolamine (ratio of ethanolamine kinase to choline kinase = 0.30). This ratio remained constant throughout the purification procedure. The K m for choline (3.0 · 10 −5 M) was lower than that of ethanolamine (1.2 · 10 −3 M). Choline was also found to inhibit ethanolamine kinase activity by 50% at a concentration of 0.005 mM, while ethanolamine inhibited choline only at very high concentrations (100–150 mM). When the enzyme was subjected to inactivation by heat, hemicholinium-3, trypsin digestion, and p-hydroxymercuribenzoate, both ethanolamine kinase and choline kinase activities were destroyed at the same rate. Freezing and thawing in the absence of glycerol also destroyed both activities at the same rate. Based on these findings, we conclude that in adult African Green monkey lung tissue, there is only one enzyme for the phosphorylation of ethanolamine and choline, and that choline phosphorylation predominates.

Further studies on the comparative karyology of the African Green and Rhesus monkeys

Further studies on the comparative karyology of the African Green and Rhesus monkeys

Cultivation of dengue virus type 2 in candidate substrates for vaccine production

Dengue virus type 2 from infected human serum has been adapted to DBS-FRhL-2 and WI-38 cells which are candidate substrates for virus vaccine production. Initially, the infected DBS-FRhL-2 monolayers were refed with maintenance medium and were reharvested after prolonged incubation. With WI-38 cells, virus could be recovered only if previously passaged in DBS-FRhL-2 or primary African Green monkey kidney cells. Peak titers of 10 6 per 0·2 ml were obtained with DBS-FRhL-2 cells whereas maximum titers of 10 5 per 0·2 ml were obtained with WI-38 cells. After six passages in the two cell strains there appeared to be no attenuation of the virus in terms of mouse neurovirulence.

Isolation of Foamy Virus from Rhesus, African Green and Cynomolgus Monkey Leukocytes

Foamy virus (FV) was recovered regularly from the leukocyte of rhesus and cynomolgus monkeys and somewhat less often from African green monkey leukocytes. Virus was found in virtually all organs of experimentally infected rhesus monkeys. No illness or pathologic abnormalities were noted in these animals or in any of the naturally infected animals in spite of the prolonged period of viral persistence in various organs and tissues.

Susceptibility and immune response to experimental plague in two species of langurs and in African green (grivet) monkeys.

The susceptibility of langurs (Presbytis entellus and Presbytis cristata) and grivets (Cercopithecus aethiops) to plague infection was investigated. Langurs were more susceptible than grivets and were all uniformly infected. Sixteen of 18 P. entellus succumbed in three to 10 days to a fatal infective dose of 810-810,000 viable plague cells. All 10 P. cristata, infected sc with 81-810,000 viable cells, died within three to eight days. Eight of these 10 animals remained afebrile during the evolution of the disease; the factor(s) responsible, however, have not been explored. Eleven of 14 grivets succumbed to infective doses of 209-2,090,000 living cells of the pathogen within three to 29 days. One of four failed to become infected by 209 organisms, judging from serologic observations. Two animals from each of two groups infected with 20,900 and 209,000 organisms, respectively, survived, apparently because of individual inherent genetic resistance. Autopsy findings all resembled those of human plague. Antibody responses were demonstrable in the animals surviving infection.