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Biomedical subjects

C E Graham

Publications and source records attributed to C E Graham.

At least 37 records · Page 2Linked to original sources
PubMed1986

Urogenital challenge of primate species with Mycoplasma genitalium and characteristics of infection induced in chimpanzees.

Eighteen male and eight female primates, representing five subhuman species, were inoculated urogenitally with Mycoplasma genitalium, a microorganism recovered from men with nongonococcal urethritis. Male rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) monkeys apparently were resistant. Female squirrel monkeys (Saimiri sciureus) and female tamarins (Saguinus mystax) exhibited low-level, genital-tract infections. Male chimpanzees (Pan troglodytes) developed an obvious genital-tract infection, with some shedding organisms for 21 weeks. M. genitalium was recovered from the blood of two of the male chimpanzees, usually when large numbers of organisms were in the urethra. Female chimpanzees generally shed organisms for 12-15 weeks. Most chimpanzees colonized with the organism exhibited increased numbers of polymorphonuclear leukocytes in the genital tract and developed a significant antibody response. The results offer substantial evidence for the pathogenicity of M. genitalium for the urogenital tract of higher primates and suggest the microorganism may have a role in human genital-tract infections.

Animals↗
PubMed1984

Comparison of monoclonal antibody-based competitive and indirect enzyme-linked immunosorbent assays for the diagnosis of swine trichinosis.

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of swine trichinosis has been developed using a biotinylated monoclonal antibody and an avidin-enzyme conjugate. The assay is based on competitive binding between swine serum antibodies and a monoclonal antibody specific for an antigenic determinant present on proteins from Trichinella spiralis excretory-secretory products with molecular weights of 45,000, 49,000, and 53,000. The competitive ELISA reliably detected pigs infected experimentally with T. spiralis and eliminated false-positive reactions in pigs infected with other swine nematodes, particularly Trichurus suis. When the competitive ELISA and an indirect ELISA using affinity-isolated antigen were compared using serum from pigs with naturally-acquired infections of T. spiralis, both tests were highly effective in detecting infected animals.

Animals↗
PubMed1984

Identification and quantitation of steroids in sulfate fractions from plasma of pregnant chimpanzee, orangutan, and rhesus monkey.

Steroids in the mono- and disulfate fractions from plasma of pregnant chimpanzees (Pan troglodytes), orangutans (Pongo pygmaeus), and a rhesus monkey (Macaca mulatta) were identified by gas chromatography/mass spectrometry and quantitated by gas-liquid chromatography on open tubular glass capillary columns. Whereas the average total concentrations were 4-5 times lower, 2.3-5.5 mumol X 1(-1) vs. 10.7-19.8 mumol X 1(-1), the pattern of steroid sulfates in the chimpanzees and orangutans were very similar to that previously found in pregnant women. Twenty one steroids were identified. The 3 beta-hydroxy-5-ene steroids were the same as in humans. Saturated pregnane derivatives were predominant and increased with time during pregnancy. Four isomers each of 3-hydroxypregnan-20-one and pregnane-3,20 alpha-diol were found, having 3 alpha, 5 alpha, 3 beta, 5 beta, 3 alpha, 5 beta, and 3 beta, 5 alpha stereochemistry, respectively. The relative proportion of disulfates was slightly lower in the great apes (15-28% of the total steroid sulfates) than in humans (23-33%). The monosulfate of 5 beta-pregnane-3 alpha, 20 alpha-diol constituted 12-14% of the total in chimpanzees and 3-4% in orangutans and humans. The monosulfate of 5 alpha-pregnane-3 beta, 20 alpha-diol constituted 5-7% in chimpanzees and 11-16% in orangutans and humans, whereas the disulfate was relatively less abundant in the great apes, 4-8%, than in humans, 10-18%. Although difficult to quantitate accurately, the chromatograms indicated that the proportion of 3 beta, 5 beta-isomers was higher in great apes than in women. The presence of 5 alpha-pregnane-3 beta, 16 alpha, 20 alpha-triol and 5 alpha-pregnane-3 alpha, 20 alpha, 21-triol indicated that hydroxylations of steroid sulfates in the great apes were similar to those in pregnant women. The steroid sulfate pattern in the rhesus monkey was completely different, 3 beta-hydroxy-5-ene steroids constituting over 95% of the total. Dehydroepiandrosterone sulfate was by far the predominant steroid, followed by the disulfates of 5-androstene-3 beta, 17 beta-diol and 5-pregnene-3 beta, 20 alpha-diol and the monosulfate of 5-androstene-3 beta, 16 alpha, 17 beta-triol. The results are discussed in relation to previous knowledge of progesterone metabolism in different animal species. So far, great apes are the only species showing the same pattern of steroid sulfates in plasma as humans.

Animals↗
PubMed1984

Monoclonal antibody-purified antigen for the immunodiagnosis of trichinosis.

Monoclonal antibodies generated from spleen cells of Balb/cJ mice infected with Trichinella spiralis reacted with antigenic determinants present in some other common swine parasites, especially Trichuris suis. An antigenic determinant unique to T spiralis was recognized by one immunoglobulin M monoclonal antibody (7C2C5); this determinant was found on larval T spiralis proteins with molecular weights of 53,000, 49,000, and 45,000. These proteins were components of in vitro-derived worm excretory-secretory products. Immunoperoxidase staining techniques and reactions of the monoclonal antibody with living parasites indicated that the source of the antigen was the stichocyte. Two of the proteins (53,000 and 49,000 molecular weight) possessing this determinant were isolated by monoclonal antibody-affinity chromatography, using hybridoma 7C2C5. When used in a serologic enzyme-linked immunosorbent assay, the affinity-isolated antigen was able to completely eliminate false-positive reactions with serum from healthy swine and to detect all pigs infected with T spiralis.

Animals↗
PubMed1983

Strongyloides ratti and Trichinella spiralis: net charge of epicuticle.

The intact epicuticles of Strongyloides ratti stage-3 larvae and Trichinella spiralis stage-1 larvae were found to have a surface net negative charge. Ultrastructural studies on S. ratti using cationized ferritin and ruthenium red showed the negative charge to be dense and uniformly distributed over the epicuticular surface. Staining with acetic acid-ferric oxide hydrosol occurred at pH 1.65 and suggests that amino acid carboxyl groups were not responsible for the negative charge property. Alcian blue staining occurred at pH 0.5 and at a critical electrolyte concentration (CEC) of 0.9 M MgCl2, a property similar to that of highly sulfated mucopolysaccharides such as the proteoglycan keratan sulfate. In contrast, T. spiralis larvae failed to stain with alcian blue below pH 5.0 or at a CEC of 0.1 M, suggesting its negative charge is associated with dissociated amino acid carboxyl groups. Attempts to remove the negative charge-bearing components in the epicuticle of S. ratti by detergents, organic solvents, denaturing agents, proteases, uronidases, neuraminidases, and lipases were unsuccessful. The presence of elastin in the S. ratti larval outer cortical layer was indicated by its vulnerability to elastase and its reaction to aldehyde fuchsin-alcian blue stain. These results show that the epicuticle of S. ratti is not a typical cell membrane, although it appears to have ultrastructural similarities. It is suggested that the association of highly sulfated mucopolysaccharides with the epicuticular surface of free-living nematodes such as S. ratti L3 may reflect a greater need to protect against surface desiccation. It is also postulated that the highly negatively charged surface may have anticomplementary and anticoagulation effects.

Animals↗
PubMed1983

Menstrual cycle patterns of hormones and sexual behavior in gorillas.

Oppositely sexed pairs of gorillas were tested behaviorally during the menstrual cycle to determine the relationship between hormone concentrations of the female and the frequency of sexual activity by the pair. Five females were tested individually during two cycles with each of two males, but serum samples for hormone assay were obtained from each female only during the first cycle of testing. There was no clear relationship between hormones and behavior for the single cycle in which the serum samples were obtained, with the exception that no copulations occurred after the early luteal phase, when progesterone was greater than 5 ng/ml. Normalized behavioral data from all four test cycles for all pairs suggested that female-solicited copulations were restricted primarily to the periovulatory period. Male sexual initiative (by one of the males) accounted for most copulations temporally dissociated from the periovulatory period. Normalized hormone data for all of the females suggested that (1) attractivity was associated with estradiol concentrations during the follicular phase, (2) proceptivity with estradiol and testosterone at midcycle, whereas (3) receptivity was not associated with hormone patterns or cycle phase. The data suggest that hormones are one of several variables that contribute to the regulation of sexual behavior in gorillas.

Animals↗
PubMed1983

Diagnosis of swine trichinosis by enzyme-linked immunosorbent assay (ELISA) using an excretory--secretory antigen.

An excretory--secretory (ES) antigen was used in a serodiagnostic enzyme-linke immunosorbent assay (ELISA) for swine trichinosis. ELISA procedures included a double-antibody test, using either an anti-swine IgG or a protein A enzyme conjugate, and a triple-antibody test using a pig IgG heavy-chain specific second antibody with a conjugated third antibody. The ES antigen was effective in eliminating all false-positive reactivity in sera from farm-raised hogs. The triple-antibody procedure was more sensitive and demonstrated a greater efficiency in detecting positive animals and early seroconversions. Naturally-infected pigs with worm burdens as low as 0.01 larvae per gram (LPG) of diaphragm were seropositive using these procedures. Seroconversion in experimentally-infected animals receiving low doses of muscle larvae (500) occurred considerably later than in animals receiving high doses (10000). This might account for false-negative reactions in naturally-infected animals with very low (less than 0.1 LPG) worm burdens.

Animals↗
PubMed1983

Shedding of antibody complexes by Strongyloides ratti (Nematoda) larvae.

Antigens on the epicuticular surface of Strongyloides ratti infective third-stage larvae (L3) could be demonstrated by an indirect fluorescent antibody technique under certain conditions. Infective L3 shed anti-antibody complexes at room temperature, but not at 4 C or in the presence of sodium azide or colchicine. Shedding of antibody did not appear to involve epicuticular antigens, and only occurred when anti-rat IgG was complexed to rat anti-larval antibody. However, parasitic L3 removed from rats did not exhibit this shedding reaction, suggesting that an important developmental change in cuticle physiology occurs during the transition from a free-living existence to a parasitic mode. The ability to shed foreign objects from the epicuticle of free-living infective L3 may be a defensive or protective response to soil microorganisms.

Animals↗
PubMed1982

Evaluation of three immunodiagnostic procedures for porcine strongyloidiasis.

An indirect fluorescent antibody test (IFAT), an indirect radioimmune antibody test (IRAT), and an enzyme-linked immunosorbent antibody assay (ELISA) were compared for their efficacies in detecting antibody to Strongyloides ransomi in pigs. The greatest sensitivity was exhibited by the ELISA, using a soluble, whole larval extract. The 2 indirect antibody assays, IFAT and IRAT, using intact 3rd-stage larvae (L3) were comparable in sensitivities. The use of live L3 in the IFAT and IRAT presented technical problems, because the larvae shed antibody complexes at ambient room temperature. This could be inhibited by the addition of 20 mM of sodium azide to all working solutions or by doing the tests at 4 C. Fixation of the L3 by either formalin or heat also eliminated the shedding problem. The specificities of the ELISA and IFAT were supported by the negative results obtained with serum from pigs infected with Ascaris suum, Stephanurus dentatus, and Trichinella spiralis.

Animals↗
PubMed1981

Chimpanzee reproductive senescence: a possible model for evolution of the menopause.

Reproductive senescence was studied in 2 female Pan troglodytes and one Pan paniscus over 40 yr old. Menstrual cycle data for these animals, in the last 3 yr, when compared to that of the same animals in previous years and to records obtained between 1967 and 1980 on 51 Pan troglodytes between 18-39 yr old, demonstrated increased length of menstrual cycles, as shown by decreases in frequency of menses. Oestrone glucuronide and pregnanediol glucuronide were measured in 24-h urine samples by radioimmunoassay. The pattern of excretion differed only slightly from that of younger animals. One Pan troglodytes had reduced oestrogen levels, but normal gonadotropin levels. The single ovulatory peak of LH normally seen in younger animals was replaced by multiple peaks in the Pan troglodytes. Menopause was observed in the Pan paniscus as documented by cessation of menstrual cyclicity, elevation of gonadotropin levels, and exaggerated response to injection of 100 microgram of gonadotropin-releasing hormone. The FSH: LH ratio was reversed in this animal. Histology of the ovaries of the Pan paniscus revealed a paucity of primary and developing follicles, and an increase in fibrous tissue. The Pan troglodytes appear to be peri-menopausal. This data suggests that the chimpanzee may serve as a model for certain phases of reproductive senescence in the human.

Aging↗
PubMed1981

The intermetatarsal nerve: a microscopic evaluation.

Microscopic evaluation of the interdigital neuroma is difficult because the appearance of the normal nerve has not been described precisely. In an effort to characterize the normal interdigital nerve, 104 nerves were removed at autopsy from the intermetatarsal spaces of 14 women who were more than 48 years old. A positive relationship between perineurial width and internal diameter of the nerve fascicle in web spaces 2, 3, and 4 was defined. In addition, the number and the diameter of blood vessels had a positive correlation in all web spaces. These measurements should be of help in the objective differentiation of the common interdigital neuroma from the normal interdigital nerve.

Aged↗
PubMed1979

Reproductive function in aged female chimpanzees.

Reproductive function was evaluated in ten female chimpanzees (Pan troglodytes) aged 35-48 years. Forty-eight years is the longevity record for the chimpanzee. Data on cycle frequency and duration was available for seven animals. Most were cycling regularly until death, and all had experienced at least one menstrual cycle within one year of death. After exclusion of periods when the animals were pregnant or in postpartum amenorrhea, the mean cycle frequency (+/- standard error)/year was 9.54 +/- 0.20 in seven animals aged 15-25 (432 cycles analyzed) compared to 8.6 +/- 0.76 in the same animals at age 35 + years (405 cycles analyzed); this effect approached significance (p = 0.072, Mann-Whitney U-test). Cycle length of 16 cycles in each of seven animals aged 15-25 was 32.23 +/- 0.38 days. The same animals when aged over 35 had mean cycle lengths of 35.59 +/- 0.73. This difference was not significant, although cycle lengths clearly increased with age in some individual animals. In five aged animals for which mating data was available, appropriate exposure to a male occurred in 52 cycles, but only two pregnancies occurred; one pregnancy resulted in a live birth at age 38, the other in a stillbirth at age 40. This conception rate was 3.85% compared with 20% in the same animals aged 15-25. These data suggest greatly reduced fertility after age 35, although menstrual cycle frequency remained high. The persistence of menstrual cyclicity until death, which occurred due to natural causes at latest in the fifth decade, is in striking contrast to the human female in which menopause occurs in the fifth decade and death is often postponed for several more decades.

Aging↗