A Comparison of Endogenous Development of Three Isolates of Cryptosporidium in Suckling Mice1

ABSTRACT. Suckling mice were used as a model host to compare the endogenous development of three different isolates of Cryptosporidium: one from a naturally infected calf, one from an immunocompetent human with a short-term diarrheal illness, and one from a patient with acquired immune deficiency syndrome (AIDS) and persistent, life-threatening, gastrointestinal cryptosporidiosis. After oral inoculation of mice with oocysts, no differences were noted among developmental stages of the three isolates in their sites of infection, times of appearance, and duration, morphology, and fine structure. Sporozoites excysted within the lumen of the duodenum and ileum, penetrated into the microvillous region of villous enterocytes, and developed into type I meronts with six or eight merozoites. Type I merozoites penetrated enterocytes and underwent cyclic development as type I meronts or they became type II meronts with four merozoites. Type II merozoites did not exhibit cyclic development but developed directly into sexual forms. Microgamonts produced £16 small, bullet-shaped microgametes, which were observed attaching to and penetrating macrogametes. Approximately 80% of the oocysts observed in enterocytes had a thick, two-layered wall. After sporulating within the parasitophorous vacuole, these thick-walled oocysts passed through the gut unaltered and were the resistant forms that transmitted the infection to a new host. Approximately 20% of the oocysts in enterocytes consisted of four sporozoites and a residuum surrounded only by a single oocyst membrane that ruptured soon after the parasite was released from the host cell. The presence of thin-walled, autoinfective oocysts and recycling of type I meronts may explain why a small oral inoculum can produce an overwhelming infection in a suitable host and why immune deficient persons can have persistent, life-threatening cryptosporidiosis in the absence of repeated oral exposure to thick-walled oocysts.     

Fine Structure of Human Malaria In Vitro*†

SYNOPSIS. The erythrocytic cycle of the human malaria parasite, Plasmodium falciparum, was examined by electron microscopy. Three strains of parasites maintained in continuous culture in human erythrocytes were compared with in vivo infections in Aotus monkeys. The ultrastructure of P. falciparum is not altered by continuous cultivation in vitro. mitochondria contain DNA-like filaments and some cristae at all stages of the erythrocytic life cycle. The Golgi apparatus is prominent at the schizont stage and may be involved in the formation of rhoptries. In culture, knob-like protrusions first appear on the surface of trophozoite-infected erythrocytes. The time of appearance of knobs on cells in vitro correlates with the life cycle stage of parasites which are sequestered from the peripheral circulation in vivo. Knob material of older parasites coalesces and forms extensions from the erythrocyte surface. Some of this material is sloughed from the host cell surface. The parasitophorous vacuole membrane breaks down in erythrocytes containing mature merozoites both in vitro and in vivo. Merozoite structure is similar to that of P. knowlesi. The immature gametocytes in culture have no knobs.     

PRIMARY PEPTIDE SEQUENCES FROM SQUID MUSCLE AND OPTIC LOBE MYOSIN IIs: A STRATEGY TO IDENTIFY AN ORGANELLE MYOSIN

The squid giant axon provides an excellent model system for the study of actin-based organelle transport likely to be mediated by myosins, but the identification of these motors has proven to be difficult. Here the authors purified and obtained primary peptide sequence of squid muscle myosin as a first step in a strategy designed to identify myosins in the squid nervous system. Limited digestion yielded fourteen peptides derived from the muscle myosin which possess high amino acid sequence identities to myosin II from scallop (60–95%) and chick pectoralis muscle (31–83%). Antibodies generated to this purified muscle myosin were used to isolate a potential myosin from squid optic lobe which yielded 11 peptide fragments. Sequences from six of these fragments identified this protein as a myosin II. The other five sequences matched myosin II (50–60%, identities), and some also matched unconventional myosins (33–50%). A single band that has a molecular weight similar to the myosin purified from optic lobe copurifies with axoplasmic organelles, and, like the optic lobe myosin, this band is also recognized by the antibodies raised against squid muscle myosin II. Hence, this strategy provides an approach to the identification of a myosin associated with motile axoplasmic organelles.     

Changes in the Architecture of Cancellous Bone in the Femora of Developing Mice as a Result of Short Duration Hypergravity

SYNOPSIS. Mice were used as a model system to demonstrate theeffects of increased loading on the trabecular bone in the developingproximal femur. A two-dimensional free body diagram revealedthat because the femur is normally held in a horizontal positionduring support, functional loading resulted in a net posteriorbending moment and a muscle generated axial compressive loadthat was 7.7 times the normal support weight for that limb.Experimental treatment consisted of four times normal gravityfor 10 min, six times per day for 30 days in addition to unrestrainednormal exercise. Unrestrained normal exercise served as thecontrol. An animal whose foot was amputated by his mother shortlyafter birth was used as a less than normal load example forcomparison with the hypergravity and control groups. All groupswere treated for 30 days. Two areas of trabecular bone wereexamined: a proximal central webbing area that may be underpredominantly tensile loading much of the time, and a more distalarea near the third trochanter that presumably experiences bendingstresses when the animal is exposed to normal activity or hypergravity.Results indicate that: 1) The central webbing area appears tohave a similar mass of bone regardless of loading conditions;however, with increased loading the trabeculae become orientedin directions that approximate directions of presumed principaltensile stresses. 2) There is no genetically predetermined amountof bony tissue in the third trochanter area; when the limb isrelatively unloaded no trabeculae are present, as the loadingincreases trabecular bone is formed. In the hypergravity group,trabeculae in the area of the third trochanter increased inlength and complexity and became more centrally positioned inthe lumen of the femur, a position that presumably would allowthe femur to better resist bending moments. These results demonstratethat short duration, high intensity functional loading can stimulategrowth of trabecular bone, and further, specific types of functionalloading can be correlated with specific architectural changes.     

Behavioral Adaptations of Intertidal Hermit Grabs

SYNOPSIS. AS an ecotone, the littoral environment is often complexand is generally rich in numbers and species of organisms. Thedisadvantages of the biotope in terms of exposure to physicalfactors of both the marine and terrestrial environments arepatent, but the advantages are not so evident. The continualreplenishment of food brought from the sea, particularly fordetritus-feeding animals such as hermit crabs, coupled withthe possibility and ability to establish microhabitatswith microclimaticconditions may constitute the principal advantage. Escape fromspecialized predators may also be important. Hermit crabs havesuccessfully exploited most intertidal environments. As membersof the "benthic detritus-feeding guild" food is abundant, andby utilizing their shells in conjunction with movements withinthe littoral zone they have met successfully most of the rigorsof the environment. The shell also provides some protectionfrom predation, particularly from non-specialized predators.Indeed,the behavioral patterns associated with living in shells whichpermit the shell to serve as a microhabitat constitute the majoradaptation enabling the hermit crabs to exploit the intertidalenvironment so successfully.     

Enzymatic hydrolysis of soluble cellulose derivatives as measured by changes in viscosity

Observation of changes in fluidity is presented as a method for following the enzymatic hydrolysis of soluble cellulose derivatives. The activity of different cell-free enzyme preparations may be compared by this method, providing certain precautions are observed. In general, results obtained by use of the fluidity method are similar to those obtained using the reducing sugar technique, indicating that the same enzyme system is measured by the two methods. Changes in the DP of the substratum may be followed within certain limits of molecular size. Results indicate that a random splitting of CMC occurs during enzymatic hydrolysis, with a concomitant decrease in intrinsic viscosity and an increase in reducing sugars. Certain inadequacies of the cellulose-cellobiose-glucose theory, together with more recent findings, have led to the postulation of an alternate explanation of the mechanism of cellulose hydrolysis.