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Biomedical subjects

C R Sterling

Publications and source records attributed to C R Sterling.

At least 73 records · Page 4Linked to original sources

Experimental infection of Aotus monkeys with Plasmodium falciparum Light and electron microscopic changes.

Six splenectomized Aotus owl monkeys were infected with P. falciparum. The animals were sacrificed when they became moribund and tissues were faken for light and electron microscopic studies. Electron microscopy revealed marked structural changes in both the liver and the heart. In the liver there was glycogen depletion, lipid droplet infiltration mitochondrial disorganization with reduction in numbers and alterations of the microvilli in the sinusoidal pole and the bile canaliculi. In the heart, there was lipid droplet infiltration and focal disorganization of the myofibrils around the intercalated disc. There was focal mitochondrial disorganization and mitochondrial concentration around the muscle lesion. The ultrastructural changes in liver and heart were similar to those described in animals dying of shock, of at least two hours duration, in different experimental models. Ultrastructural lesions found in other organs were less marked. In the kidneys there was mainly a moderate lipid infiltration of the tubular epithelium. This change is probably the early precursor of that seen at four hours in animals dying in experimental shock. The above changes are discussed in light of the known pathophysiology of both experimental infections in animals and natural human malarial infections.

Animals↗

Erythrocyte membrane alterations induced by Plasmodium simium infection in Saimiri sciureus: relation to Schüffner's dots.

The nature of erythrocyte membrane alterations in Plasmodium simium infections was determined employing light microscopy, carbon replication and transmission electron microscopy. Light microscopy of Giemsa stained preparations shows that infected cells initially acquire a faint stippling (schuffnerization) which becomes pronouced with subsequent parasite development. Enlargement of the host cell usually accompanied stippling. Both phenomena appear to depend on host cell age since infected mature erythrocytes were neither stippled nor enlarged. Carbon replicas show numerous indentations over the outer membrane surface of most infected cells. Their distribution suggests that they account for Schuffner's granules. The surface indentations are manifest as small infundibular which open to the infected cell's surface. Cytoplasmic microvesciles in the infected cell's stroma frequently are observed adjacent or catenated to the surface infundibula. Images suggest their funsion with the surface infundibula thus adding membrane to the cell's surface and accounting for host cell enlargement.

Animals↗

Comparison of the host ranges and antigenicity of Cryptosporidium parvum and Cryptosporidium wrairi from guinea pigs.

Oocysts of a Cryptosporidium isolate from guinea pigs were not infectious for adult mice, but were infectious for two of three newborn calves and for suckling mice. However, oocysts isolated from calves or mice infected with guinea pig Cryptosporidium were not infectious for guinea pigs. Four isolates of C. parvum from calves were incapable of infecting weanling guinea pigs. Microscopic examination of tissue from the colon and cecum of suckling guinea pigs inoculated with C. parvum revealed sparse infection of some pups. These host range studies and previously described differences in 125I-labeled oocyst surface protein profiles between Cryptosporidium sp. from guinea pigs and C. parvum suggest they are distinct species. We propose the name Cryptosporidium wrairi be retained. Studies with monoclonal antibodies indicate that C. wrairi and C. parvum are antigenically related.

Animals↗

Cyclospora: an enigma worth unraveling.

In part, Cyclospora cayetanensis owes its recognition as an emerging pathogen to the increased use of staining methods for detecting enteric parasites such as Cryptosporidium. First reported in patients in New Guinea in 1977 but thought to be a coccidian parasite of the genus Isospora, C. cayetanensis received little attention until it was again described in 1985 in New York and Peru. In the early 1990s, human infection associated with waterborne transmission of C. cayetanensis was suspected; foodborne transmission was likewise suggested in early studies. The parasite was associated with several disease outbreaks in the United States during 1996 and 1997. This article reviews current knowledge about C. cayetanensis (including its association with waterborne and foodborne transmission), unresolved issues, and research needs.

Animals↗

Restriction fragment length polymorphism analysis of Cryptosporidium parvum isolates of bovine and human origin.

Cryptosporidium parvum oocysts isolated from different hosts and geographical areas were compared by restriction endonuclease analysis of repetitive DNA: Iowa (bovine), Florida (bovine), New York (bovine), Peru (human), Brazil (human), and Mexico (human). Southern blot hybridization analysis was performed using the restriction endonuclease enzyme Eco RI and the DNA probe pV47-2. The probe hybridized with 18 bands present in all the isolates. The Brazilian, Mexican, and Peruvian human isolates had an additional common band of 4.3 kbp that was absent in the bovine isolates. Two extra bands of 14 and 12 kbp were present in the Brazilian isolate whereas the Mexican isolate had an extra band of 14 kbp. When the Iowa and Peru C. parvum isolates were passed twice through calves, oocysts recovered from both passages showed identical banding patterns, suggesting that recombination of the repetitive sequences was not altered during sexual reproduction. The DNA digested with other restriction endonucleases were tested confirming differences between isolates. A genomic DNA library is currently being produced to better define isolate variation in C. parvum.

Animals↗

Cryptosporidium infections in inbred strains of mice.

Cryptosporidium, a protozoan parasite of man and animals, is an important etiological agent of diarrhea throughout the world, particularly in children and immunocompromised individuals such as AIDS patients. Unfortunately, because of the lack of both in vivo laboratory models and reliable in vitro parasite culture systems, virtually nothing is known about the immunological events occurring during disease. In order to identify reliable animal models for infection, we studied C. parvum infections in 19 different strains of mice representing 12 H-2 haplotypes: A/J, AKR/J, B10.D2/J, B10.M/J, C3H/HeJ, C57BL/65, C57BL/6J-bgJ, CBA/NJ, DBA/1J, DBA/2J, HRS/J, HTG/J, NZB/B1NJ, NZW/J, P/J, RIII/J, SJL/J, SWR/J, and WB/ReJ, and in one gerbil: Meriones unguiculatus. Fecal samples and histological sections of the intestine taken on day 7 post-Cryptosporidium inoculation indicated that only the beige mouse (C57BL/6J-bgJ) harbored significant numbers of parasites compared to the other strains. The numbers of parasites harbored in these NK cell-deficient beige mice were, however, considerably lower than those seen in neonatal mice. Adult inbred mouse strains susceptible to Cryptosporidium infections are discussed.

Animals↗

Susceptibility dynamics in neonatal BALB/c mice infected with Cryptosporidium parvum.

BALB/c Mice were infected as neonates and at different ages to study the susceptibility dynamics in this animal model to Cryptosporidium parvum. When 4-day-old animals were infected with 10(5) C. parvum oocysts, parasites were detected in the terminal ileum when the mice became 14-25 days old (10-21 days post-infection [PI]). The percentage of animals positive for parasites was 100% up to the age of 19 days (15 days PI) but decreased immediately thereafter until no parasites were detected in 26-day-old (22 days PI) or older mice. Parasite load also decreased in these animals from 184.7 parasites per high power field in 14-day-old animals (10 days PI) to 0.22 in 25-day-old (21 days PI) mice. In a second study, some neonatal mice became resistant to C. parvum when infection was attempted at day-10 of age (day-15 of age at sacrifice). The susceptibility to C. parvum decreased until 14 days of age (19 days of age at sacrifice) when mice could no longer be infected. Parasite load also decreased in infected mice from 235.6 parasites per high power field (9 days of age at sacrifice) to 0.25 (18 days of age at sacrifice).

Aging↗

Cryptosporidium merozoite isolation and purification using differential centrifugation techniques.

Simple modifications to a recently published merozoite purification procedure (Bjorneby et al., J. Immunol. 145:298, 1990) increased yields 3- to 5-fold. Calves were infected with 2.5 x 10(8) Cryptosporidium parvum oocysts and sacrificed 65 h post-infection. The ilium and caecum were removed. The tissue was sieved through a large strainer (2 mm2) to produce a homogeneous suspension. Red blood cells were removed by differential centrifugation (600 g); merozoites remained in the supernatant. The merozoites were pelleted (2,100 g) and washed in modified Hank's balanced salt solution deficient in Mg+2 and Ca+2. Percoll purification (density 1.070 g/ml and centrifugation speed of 22,000 g for 30 min) yielded 8 x 10(8) merozoites. Nineteen monoclonal antibodies (MAb) detected by either an enzyme-linked immunosorbent assay or an immunofluorescence assay, have been generated against the merozoite stage. Gels of proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver-stained showed that sporozoites and merozoites have many common lower molecular weight proteins. Western blots of sporozoite and merozoite antigens reacted with anti-sporozoite MAb showed several cross-reacting antigens shared by these life-cycle stages.

Animals↗

The role of breast milk in protecting urban Peruvian children against cryptosporidiosis.

To test the hypothesis that breast milk of nursing mothers may afford children protection against cryptosporidiosis, a prospective cohort study was carried out in the young peoples' community of San Juan de Miraflores near Lima, Peru. Mothers and newborn children were sorted into cohort groups based on the mothers' breast milk antibody response to Cryptosporidium sporozoites using an antibody-capture enzyme-linked immunosorbent assay to detect parasite-specific immunoglobulin A. Children were monitored for Cryptosporidium infection using an indirect immunofluorescence assay. Of 211 mothers enrolled in the study, 39 (18.5%) had high breast milk antibody titers, 107 (50.7%) had medium titers, and 65 (30.8%) had low titers. Sixty-one episodes of Cryptosporidium infection were detected in 50 children of these mothers. Eleven (22%) had mothers in the high antibody titer group, 20 (40%) had mothers in the medium titer group, and 19 (38%) had mothers in the low titer group. The prevalence of infection within children of each group was 0.17, 0.19 and 0.38 respectively. There was no significant difference in the prevalence or duration of infection among children of the different groups. The data does not support the notion that there is protection from Cryptosporidium infection afforded children whose mothers have demonstrable breast milk antibodies against the parasite.

Adult↗

Hyperimmune hens as a novel source of anti-Cryptosporidium antibodies suitable for passive immune transfer.

Leghorn hens were subcutaneously immunized with 25 micrograms of Cryptosporidium parvum oocyst emulsified in Freund's complete adjuvant. A booster dose was injected 5 weeks later. Anti-Cryptosporidium activities of yolks and sera measured by an enzyme-linked immunosorbent assay (ELISA), demonstrated high levels in both sera and egg yolks which persisted for at least 17 wk. Preparations from yolks with high, medium and low anti-Cryptosporidium ELISA activities were used in a neonatal mouse model to assess their biological activities. A significant parasite reduction (P less than or equal to 0.001) was found between the high and all other groups. Hyperimmune eggs could be used as a source for passive immunity in cryptosporidiosis.

Animals↗