Concanavalin a capping of rhesus monkey polymorphonuclear leukocytes

The technique of capping, a probe designed to evaluate the fluidity and functional competence of human polymorphonuclear leukocytes (PMNs), has been successfully adapted for rhesus monkey PMNs. The capping characteristics of rhesus monkey PMNs are very similar to those of human PMNs. Utilizing this capping technique as an evaluation tool and with fetal/neonatal rhesus monkey as a functional animal model, the ontogeny of movement and chemotactic characteristics of PMNs can now be studied.     

Laparoscopic studies of ovulation, pregnancy diagnosis, and follicle aspiration in sheep

A technique for rapid and repeated laparoscopic ovarian examination was developed for use in the sheep. A midventral approach was used on ewes under general anesthesia on a 45-degree sloped table. Examination could be done as often as every other day.The technique was used for detection of ovulation in studies of gonadotropin stimulation in anestrous ewes. Ovulation was induced with 5 to 10 mg FSH and 100 to 2000 iu HCG. The use of laparoscopy in pregnancy diagnosis was also investigated. Accurate diagnosis could be made by Day 17 using both uterine and ovarian characteristics. Visualization through the laparoscope was utilized for aspiration of follicular oocytes using a 23 ga, 11 cm needle.     

Effect of tunicamycin on cell fusion induced by Mason-Pfizer monkey virus.

Mason-Pfizer monkey virus, a D-type retrovirus, has been shown to induce multinucleate cell (syncytium) formation or cell fusion in several normal primate cells. A series of experiments has been carried out to examine whether a glycosylated "fusion-inducing" product is responsible for this biological property of Mason-Pfizer monkey virus. Treatment of rhesus monkey fetal lung cells with different concentrations of tunicamycin, a potent inhibitor of glycosylation, during infection with Mason-Pfizer monkey virus had no effect on cell fusion even though up to 5 micrograms of the drug per ml was tested. Furthermore, no significant effect on the extent of syncytium formation in rhesus monkey fetal lung cells was observed when the time of addition or duration of treatment with this inhibitor was varied. Nevertheless, tunicamycin was very effective in blocking glycosylation in rhesus cells since virions produced in the presence of this drug completely lacked gp70 and gp20, the two structural glycoproteins of Mason-Pfizer monkey virus. These non-glycosylated virus particles produced in the presence of tunicamycin were noninfectious as determined by a protein A binding assay and were unable to induce syncytium formation when assayed on rhesus cells. These results indicate that glycosylation of the fusion-inducing product is not required for multinucleate cell formation induced by Mason Pfizer monkey virus.     

Development and application of ovine reproductive technologies: an Indian experience

Sheep play an important role in the Indian economy by providing employment to a large population of marginal and landless farmers. The production from native breeds is relatively low due to their poor reproductive efficiency. Embryo transfer technology can be utilised for faster multiplication of elite animal to increase the genetic gain. A great deal of research is involved to overcome the constraints in the technology, i.e. expensive and complicated nature of the technology and low success rate. In order to avoid surgical involvement, procedures of laparoscope aided embryo collection and transfer have been developed. Although the use of FSH of ovine origin has given more consistent superovulatory response, but it is still too low to get sufficient numbers of progeny from a donor ewe. The progress made in cryopreservation of ram semen has opened the possibility for conservation and utilisation of frozen semen of elite rams in sheep improvement programme. The lambing rate obtained after laparoscope aided intrauterine artificial insemination with frozen semen is encouraging but the impetus is now to develop the non-invasive transcervical insemination technique.     

Identification of postentry restrictions to Mason-Pfizer monkey virus infection in New World monkey cells

TRIM5α has been shown to be a major postentry determinant of the host range for gammaretroviruses and lentiviruses and, more recently, spumaviruses. However, the restrictive potential of TRIM5α against other retroviruses has been largely unexplored. We sought to determine whether or not Mason-Pfizer monkey virus (M-PMV), a prototype betaretrovirus isolated from rhesus macaques, was sensitive to restriction by TRIM5α. Cell lines from both Old World and New World primate species were screened for their susceptibility to infection by vesicular stomatitis virus G protein pseudotyped M-PMV. All of the cell lines tested that were established from Old World primates were found to be susceptible to M-PMV infection. However, fibroblasts established from three New World monkey species specifically resisted infection by this virus. Exogenously expressing TRIM5α from either tamarin or squirrel monkeys in permissive cell lines resulted in a block to M-PMV infection. Restriction in the resistant cell line of spider monkey origin was determined to occur at a postentry stage. However, spider monkey TRIM5α expression in permissive cells failed to restrict M-PMV infection, and interference with endogenous TRIM5α in the spider monkey fibroblasts failed to relieve the block to infectivity. Our results demonstrate that TRIM5α specificity extends to betaretroviruses and suggest that New World monkeys have evolved additional mechanisms to restrict the infection of at least one primate betaretrovirus.     

Routine measurement of urinary total estrogens of the female Japanese monkey as an index for estimating the time of ovulation

The fluorometry of urinary total estrogens ofBrown et al. (1968) has been modified and successfully applied to the routine measurement of urinary estrogens in Japanese monkeys. Serial analysis during the menstrual cycle of the Japanese monkey has shown typical changes in urinary total estrogens with a distinct midcycle peak which preceded the day of ovulation by 0–2 days. The technique is valuable in an accurate estimation of the time of ovulation for successful programmed breeding of the Japanese monkey.     

Rapid oral endotracheal intubation with a fibre-optic scope in rabbits: a simple and reliable technique

The orotracheal intubation of rabbits is complicated by their oropharyngeal anatomy. Numerous techniques to intubate rabbits have been described; however, these methods require specialized devices, tracheostomy, or are performed in a blind fashion. We describe a technique for the intubation of the rabbit under direct visualization with a standard, small-bore, fibre-optic laparoscope, which is both rapid and simple to perform.     

Dynamics of primate bipedal walking as viewed from the force of foot

Bipedal walking of the six species of anthropoid primates including man were examined by means of the force plate technique. Though each species has a particular pattern of bipedal walking, we can classify two types of patterns in these primates as far as the foot force is concerned. The first type includes the man, chimpanzee, and spider monkey and the second type contains the Japanese monkey, hamadryas baboon, and gibbon. It was emphasized that the similarity of man to the chimpanzee and spider monkey in bipedal walking has some evolutionary significance.     

Restricted changes in the adenovirus DNA-binding protein that lead to extended host range or temperature-sensitive phenotypes

Human adenovirus fails to multiply efficiently in monkey cells owing to a block to late viral gene expression. Ad2hr400 through Ad2hr403 are a set of host range (hr) mutants which were selected for their ability to readily grow in these cells at 37 degrees C. The mutations responsible for this extended host range have previously been mapped to the 5' portion of the gene encoding the 72-kilodalton DNA-binding protein (DBP). DNA sequence analyses indicate that all four hr mutants contain the same alteration at coding triplet 130, which changes a histidine codon to a tyrosine codon. These results extend those of Anderson et al. (J. Virol. 48:31-39, 1983), which suggested that only this change in the DBP amino acid sequence can expand adenovirus host range to monkey cells. The hr phenotype does not appear to require phosphorylation of this tyrosine residue, since no phosphotyrosine was detected in DBP isolated from Ad2hr400-infected monkey cells. The hr mutants Ad2hr400 through Ad2hr403, however, are cold sensitive for growth in monkey cells. The mutant Ad2ts400, which was derived from Ad2hr400, represents a second class of hr mutants which can grow efficiently in monkey cells at 32.5 degrees C. The cold-resistant hr mutation of Ad2ts400 has previously been mapped to the 5' region of the DBP gene (map units 63.6 through 66). DNA sequence analysis of this region shows that this mutant contains the original hr alteration at coding triplet 130 as well as a second alteration at coding triplet 148, which changes an alanine codon to a valine codon. We suspect that the alterations at amino acids 130 and 148 change the structure of the amino-terminal domain of the DBP, allowing it to better interact with monkey cell components required for late viral gene expression. Ad2ts400 also contains a temperature-sensitive mutation which has previously been mapped to the 3' portion of the DBP gene (map units 61.3 through 63.6). Sequence analysis of this region indicates that the DBP coding triplet 413 has been altered. This change from a serine codon to a proline codon is the same alteration reported in the previously sequenced DBP mutants Ad5ts125 (W. Kruijer et al., Nucleic Acids Res. 9:4439-4457, 1981) and Ad5ts107 (W. Kruijer et al., Virology 124:425-433, 1983). Thus it appears that only a very limited number of changes in either the 5' or the 3' portion of the DBP gene can give rise to the hr or temperature-sensitive phenotypes, respectively.     

Biochemical purification of monkey and human LDH isoenzyme 5 suitable as immunization antigen

A method consisting in an ion-exchange chromatography on DEAE-Sephadex A50 and an affinity chromatography on Oxamate-hexyl-Sepharose for the purification of monkey and human LDH isoenzyme 5 is presented. Analytical isoelectric focusing reveals minor variants in the monkey LDH isoenzyme 5. This material has been used to produce an antiserum from rabbits which recognizes all the human LDH isoenzymes containing the M subunit, without cross-reacting with LDH1.     

Linkage relationship between the genes for thymidine kinase and galactokinase in different primates.

In this study we investigated the expression of primate galactokinase in somatic cell hybrids between a thymidine kinase-deficient mouse cell line and two different primate cell lines, one of which was derived from African green monkey kidney cells and the other from chimpanzee fibroblasts. All the African green monkey-mouse hybrid clones, selected in HAT medium, expressed monkey galactokinase activity and contained a monkey chromosome similar to a human E-group chromosome. When these clones were backselected in medium containing 5-bromodeoxyuridine, both this chromosome and the monkey galactokinase activity were lost. All the hybrid clones between mouse and chimpanzee cells, which were selected in HAT medium, contained the chimpanzee chromosome 17 and expressed chimpanzee galactokinase activity. These results indicate that the linkage relationship between galactokinase and thymidine kinase has been maintained in 3 divergent primate species--man, chimpanzee, and Old World monkey.     

Patterns and determinants of monkey densities in Peru and Bolivia,with notes on distributions

A comparative study of species assemblages and population densities was conducted on Amazonian monkey communities in 16 areas, ranging from 3°S latitude in northern Peru to 18°S latitude in southern Bolivia. The habitats ranged from several types of tropical rain forest in the more northern latitudes to dry, deciduous forest in the southernmost study area. The monkey populations of three of the study areas have historically received light hunting pressure; the rest have been moderately to heavily hunted. A transect census technique was used to estimate the relative and absolute densities of all diurnal monkey species except Cebuella pygmaea. The number of coexisting monkey species ranged from 4–6 in the southern areas to 12–14 in the northern areas. The reduction in species richness in central and southern areas of Bolivia is probably attributable to several inimical habitat factors. Predation by humans was found to be the single most important factor affecting monkey densities. Monkey densities, and especially biomasses, were much lower in areas not protected from hunting than in protected areas. Hunting did not affect all species equally. Larger-sized species are hunted more and have severely reduced numbers in unprotected areas, whereas the densities of smaller species are not noticeably diminished in unprotected areas. Large, herbivorous monkey species contributed the major proportion of the total monkey biomass in protected areas. The strong influence of hunting has largely obscured the effects of other factors on population densities.     

Comparison of oxime reactivation and aging of nerve agent-inhibited monkey and human acetylcholinesterases

Non-human primates are valuable animal models that are used for the evaluation of nerve agent toxicity as well as antidotes and results from animal experiments are extrapolated to humans. It has been demonstrated that the efficacy of an oxime primarily depends on its ability to reactivate nerve agent-inhibited acetylcholinesterase (AChE). If the in vitro oxime reactivation of nerve agent-inhibited animal AChE is similar to that of human AChE, it is likely that the results of an in vivo animal study will reliably extrapolate to humans. Therefore, the goal of this study was to compare the aging and reactivation of human and different monkey (Rhesus, Cynomolgus, and African Green) AChEs inhibited by GF, GD, and VR. The oximes examined include the traditional oxime 2-PAM, two H-oximes HI-6 and HLo-7, and the new candidate oxime MMB4. Results indicate that oxime reactivation of all three monkey AChEs was very similar to human AChE. The maximum difference in the second-order reactivation rate constant between human and three monkey AChEs or between AChEs from different monkey species was 5-fold. Aging rate constants of GF-, GD-, and VR-inhibited monkey AChEs were very similar to human AChE except for GF-inhibited monkey AChEs, which aged 2-3 times faster than the human enzyme. The results of this study suggest that all three monkey species are suitable animal models for nerve agent antidote evaluation since monkey AChEs possess similar biochemical/pharmacological properties to human AChE.     

Identification of New World monkey MHC-DRB alleles using PCR,DGGE and direct sequencing

Identification of New World monkey MHC-DRB alleles has previously relied upon labor-intensive cloning and sequencing techniques. Here we describe a rapid and unambiguous way to distinguish DRB alleles in New World monkeys using the polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), and direct sequencing. The highly variable second exon of New World monkey DRB alleles was amplified using generic DRB primers and alleles were separated by DGGE. DNA was then reamplified from plugs removed from the gel and alleles were determined using fluorescent-based sequencing. The validity of this typing procedure was confirmed by the identification of all DRB alleles previously characterized by cloning and sequencing techniques from an individual cotton-top tamarin. Importantly, our analysis revealed DRB alleles not previously identified in this reference animal. Following validation of our technique, the protocol was employed for the characterization of MHC-DRB alleles in four other species of New World monkey: the pygmy marmoset, white-faced saki monkey, long-haired spider monkey and owl monkey. Using this technique, we identified five alleles from the cotton-top tamarin, five alleles from the owl monkey, three alleles from the long-haired spider monkey, three alleles from the white-faced saki monkey and two alleles from the pygmy marmoset. On the basis of phylogenetic tree analyses, 13 new DRB alleles were assigned to eight different MHC-DRB lineages. Whereas traditional DRB typing via cloning and sequencing provides limited information, our new technique provides a simple and relatively rapid way of identifying New World monkey MHC-DRB alleles.Nucleotide sequence data reported are available in the GenBank/EMBL/DDBJ databases under the accession numbers AJ544165–AJ544177     

Late DNA replication in rhesus monkey (Macaca mulatta) chromosomes

A study of the pattern of late DNA replication in rhesus monkey chromosomes showed evident similarities with man. This must be a consequence of the evolutionary conservation of replication patterns in primate chromosomes, as it has been demonstrated in the great apes, in Cebus, and man. However, the pattern of late replication of the allocyclic X chromosome in lymphocytes of female rhesus monkey was identical with the fibroblast pattern in man, and with the pattern found in only 5 to 20% of human lymphocytes.     

Mutations that allow human Ad2 and Ad5 to express late genes in monkey cells map in the viral gene encoding the 72K DNA binding protein.

Five host-range mutants (Ad2hr400–hr403, Ad5hr404) of human adenovirus serotype 2 and 5 (Ad2 and Ad5) which overcome the block to growth of wild-type adenovirus in monkey cells have been isolated. They form plaques and multiply efficiently in both monkey and human cells. The alteration in each of these mutants allows the full expression of all viral late genes, in marked contrast to the depressed synthesis of many late proteins in monkey cells infected with the parental Ad2 or Ad5. The altered gene encodes a diffusible product, since the mutation acts in trans to enhance the synthesis of wild-type Ad3 late proteins during co-infections of monkey cells with Ad2hr400 and Ad3. Restriction enzyme analysis of the genomes of all the host-range mutants show that none of them contain major alterations. In addition, an earlier report (Klessig and Hassell, 1978) indicated that Ad2hr400 does not contain SV40 sequences, which in some adenovirus-SV40 hybrid viruses allows efficient multiplication in monkey cells. The mutation responsible for the extended host range has been physically mapped by marker rescue experiments using isolated restriction enzyme fragments of the mutants to transfer the new phenotype to wild-type adenovirus. The alteration in each of the five mutants is located in a region (coordinates 62–70.7; coordinates 62–68 for Ad5hr404) which encodes predominantly the 72K DNA binding protein. More detailed mapping using Ad2hr400 fragments places the mutation (coordinates 62.9–65.6) entirely within the 72K gene. The multifunctional nature of the 72K protein and some of its similarities to SV40 T antigen are discussed.     

Interspecies conservation of structure of interphotoreceptor retinoid-binding protein. Similarities and differences as adjudged by peptide mapping and N-terminal sequencing.

Structural properties of the retinal extracellular-matrix glycolipoprotein interphotoreceptor retinoid-binding protein (IRBP) from human, monkey and bovine retinas have been compared. SDS/polyacrylamide-gel-electrophoretic analysis of limited tryptic and Staphylococcus aureus-V8-proteinase digests show virtually identical patterns for the monkey and human proteins, whereas both sets differ considerably from the bovine protein pattern. Time-course digestion shows monkey IRBP to be more readily cleaved than bovine IRBP and also cleaved to smaller fragments. Also, reversed-phase h.p.l.c. of complete tryptic digests of the IRBPs indicate that, although they have in common a similar preponderance of hydrophobic peptides, all three proteins differ extensively in their fine structure. The N-terminal sequences of monkey and bovine IRBPs have been extended beyond those presented in our previous report [Redmond, Wiggert, Robey, Nguyen, Lewis, Lee & Chader (1985) Biochemistry 24, 787-793] to over 30 residues each. The sequences yet show extensive homology, differing at only two positions, although the major monkey sequence has an additional five amino acid residues at its N-terminus ('n + 5' sequence) not observed with bovine IRBP ('n' sequence). The newly determined N-terminal sequence of human IRBP demonstrates the presence of equal amounts of the 'n' and 'n+5' sequences that are qualitatively identical with those of the monkey. The presence of the five-amino-acid-residue extension in primate, but not bovine, IRBP may indicate variation in post-translational processing.     

Circulating blood volume of the Japanese monkey (Macaca fuscata) by means of the dye T-1824

The circulating blood volume has been measured in 65 healthy Japanese monkeys (Macaca fuscata) with the dye T-1824 (Evans blue). The values obtained were 83 ± 13 ml/kg in females, and 85 ± 12 ml/kg in females, almost the same as the figures reported in man.Sex difference of blood volume has been reported to be present in man, but from the data obtained here no significant difference was found in the Japanese monkey.In young monkeys aged from 1 to 5.5 years, the blood volume was 77 ± 5 ml/kg in males, and 78 ± 4 ml/kg in females. The blood volume of the monkey was found to increase with age; a significant difference was demonstrated between adult males and young males.     

Retrovirus restriction by TRIM5alpha variants from Old World and New World primates

The TRIM5alpha proteins of humans and some Old World monkeys have been shown to block infection of particular retroviruses following virus entry into the host cell. Infection of most New World monkey cells by the simian immunodeficiency virus of macaques (SIVmac) is restricted at a similar point. Here we examine the antiretroviral activity of TRIM5alpha orthologs from humans, apes, Old World monkeys, and New World monkeys. Chimpanzee and orangutan TRIM5alpha proteins functionally resembled human TRIM5alpha, potently restricting infection by N-tropic murine leukemia virus (N-MLV) and moderately restricting human immunodeficiency virus type 1 (HIV-1) infection. Notably, TRIM5alpha proteins from several New World monkey species restricted infection by SIVmac and the SIV of African green monkeys, SIVagm. Spider monkey TRIM5alpha, which has an expanded B30.2 domain v3 region due to a tandem triplication, potently blocked infection by a range of retroviruses, including SIVmac, SIVagm, HIV-1, and N-MLV. Tandem duplications in the TRIM5alpha B30.2 domain v1 region of African green monkeys are also associated with broader antiretroviral activity. Thus, variation in TRIM5alpha proteins among primate species accounts for the observed patterns of postentry restrictions in cells from these animals. The TRIM5alpha proteins of some monkey species exhibit dramatic lengthening of particular B30.2 variable regions and an expanded range of susceptible retroviruses.