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Biomedical subjects

M Aikawa

Publications and source records attributed to M Aikawa.

At least 325 records · Page 18Linked to original sources

Sporozoites of mammalian malaria: attachment to, interiorization and fate within macrophages.

Sporozoites of Plasmodium berghei and Plasmodium knowlesi, incubated in normal serum readily interact with peritoneal macrophages of mice or rhesus monkeys, respectively. Interiorization of the sporozoite requires that both serum and macrophages be obtained from an animal susceptible to infection by the malaria parasite. Serum requirements for sporozoite attachment to the macrophage are less specific. Phagocytosis is not essential for the parasites to become intracellular. Our findings indicate that active penetration of the sporozites into the macrohages does occur. Antibodies present in the serum of sporozoite-immunized mice are important in determining the fate of both the intracellular sporozoites and the macrophages containing the parasite. Sporozoites coated with antibodies degenerate within vacuoles of the macrophages, which have no morphologic alteration. Sporozoites incubated in normal serum do not degenerate within macrophages, but the parasitized macrophages become morphologically altered and are destroyed. Preliminary experiments indicate that sporozoites appear to interact with rat Kupffer cells in the same way as with the peritoneal mouse macrophages. It is postulated that Kupffer cells play a dual role in sporozoite-host cell interaction. In normal animals these cells might serve to localize the sporozoites in the immediate vicinity of the hepatocytes. In the immunized animals, macrophages would remove and destroy the antibody-coated parasites, thus contributing to sporozoite-induced resistance.

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Phagocytosis of Borrelia recurrentis by blood polymorphonuclear leukocytes is enhanced by antibiotic treatment.

The removal of Borrelia spirochetes from the blood in relapsing fever was studied by examining patients' blood phagocytic cells with the Dieterle silver stain. Polymorphonuclear leukocytes ingested Borrelia at increased rates for several hours after antibiotic treatment, during which time the total numbers of circulating plasma spirochetes were decreasing. Incubation of infected blood at 37 degrees C for 2 h resulted in a progressive increase in phagocytosis. Addition of penicillin G and tetracycline to infected blood caused a further enhancement of phagocytosis. Electron microscopy of polymorphonuclear leukocytes revealed spirochetes in phagosomes. These results indicated that blood polymorphonuclear leukocytes have a prominent role in removing Borrelia from the plasma and suggested that antibiotics act by altering the surface of spirochetes to render them more susceptible to phagocytosis.

Adult↗

Interaction between cytochalasin B-treated malarial parasites and erythrocytes. Attachment and junction formation.

We have previously demonstrated that invasion of erythrocytes (RBCs) by malaria merozoites follows a sequence: recognition and attachment in an apical orientation associated with widespread deformation of the RBC, junction formation, movement of the junction around the merozoite that brings the merozoite into the invaginated RBC membrane, and sealing of the membrane. In the present paper, we describe a method for blocking invasion at an early stage in the sequence. Cytochalasin-treated merozoites attach specifically to host RBCs, most frequently by the apical region that contains specialized organelles (rhoptries) associated with invasion. The parasite then forms a junction between the apical region and the RBC. Cytochalasin blocks movement of this junction, a later step in invasion. Cytochalasin-treated (Plasmodium knowlesi) merozoites attach to Duffy-negative human RBCs, although these RBCs are resistant to invasion by the parasite. The attachment with these RBCs, however, differs from susceptible RBCs in that there is no junction formation. Therefore the Duffy associated antigen appears to be involved in junction formation, not initial attachment.

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Freeze-fracture study of malaria sporozoites: antibody-induced changes of the pellicular membrane.

Plasmodium cynomolgi, Plasmodium knowlesi, and Plasmodium berghei sporozoites, before and after incubation with immune serum, were studied after freeze-fracture by electron microscopy. There were evenly distributed numerous intramembranous particles (IMP) on the P face of the outer membrane. The E face of the plasma membrane had fewer IMP than its P face. The E face of the intermediate membrane had few IMP and also linear arrays of slightly raised ridges running the length of the parasite. The P face of the intermediate membrane had many IMP aligned along the long axis of the sporozoite. On the P face of the inner membrane, IMP were arranged in very distinct rows conforming to the long axis of the parasite; the E face of this membrane had a few randomly distributed IMP. A prominent change in the sporozoite incubated in immune serum was the appearance of a layer of aggregated particles around the parasite. The P face of the plasma membrane had several clear areas devoid of IMP and IMP aggregates. No changes were seen in the other fractured faces of the pellicle. These observations suggest that immune serum acts only on the P face of the plasma membrane.

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Mesangial glomerulonephropathy with deposition of IgG, IgM, and C3 induced by mercuric chloride: a new model.

A mesangial glomerulonephropathy, characterized by the deposition of rat IgG, IgM, and C3 in the glomerular mesangium, was produced in Wistar rats by a prolonged administration of mercuric chloride (HgCl2). The HgCl2 was dissolved in sterile distilled water (0.2 mg. per ml.), and a group of 15 male Wistar rats was given injections subcutaneously three times a week on alternate days at a dosage of 0.15 mg. per 100 gm. of body weight for 27 weeks. A control group of nine rats was given injections of distilled water only. Mesangial glomerulonephropathy developed in 12 of 15 rats injected with HgCl2 and was characterized by the following: (1) coarse granular and nodular deposition of rat IgG, IgM, and C3 in the mesangium of all glomeruli, (2) absence of staining for rat albumin, IgA, and fibrin, (3) presence of electron-dense deposits in the mesangium, (4) focal and segmental proliferation of the mesangial matrix, (5) interstitial inflammation, (6) tubular atrophy, and (7) deposition of periodic acid-Schiff-positive material in the medulla adjacent to the thin limbs of the loops of Henle. Glycosuria and a slight increase in proteinuria were observed transiently in some rats. The blood urea nitrogen levels were normal in all rats. Eluates from the kidneys with heavy mesangial deposits contained rat IgG. However, the eluted antibody failed to react with normal rat kidney tissue components. None of the above findings were present in the control rats. The study provides a model of a mesangial nephropathy that seems to be immunologically induced; however, the mechanism for the formation and deposition of the immune deposits containing rat IgG, IgM, and C3, and the nature of the antigen(s) have not been elucidated.

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Electron-dense subepithelial glomerular deposits in Henoch-Schönlein purpura syndrome.

A case of Henoch-Schönlein purpura (HSP) with renal lesions is presented. The clinical, laboratory, and morphologic findings revealed features commonly described in HSP. A renal biopsy revealed diffuse, generalized, proliferative glomerulonephritis. The unusual findings in this case were numerous electron-dense subepithelial deposits ("humps") and a few small intramembranous deposits in the glomerular capillaries without deposits in the mesangium, mimicking those of acute poststreptococcal glomerulonephritis. The electron-dense deposits were well correlated with deposits of IgG, IgA, C3, C4, C1q, and fibrin-fibrinogen demonstrated by immunofluorescent microscopy.

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Mouse antibody-dependent eosinophil and macrophage adherence and damage to schistosomula of Schistosoma mansoni.

The effects of sera from S. mansoni-infected mice together with eosinophil- and macrophage-rich peritoneal exudate cells (PEC) on the schistosomula were examined in vitro. Adherence to and killing of the parasite by PEC is mediated through a complement-independent opsonizing IgG. Destruction of schistosomula was assessed morphologically and by evaluating their infectivity to normal mice. By using ferritin-labeled anti-mouse immunoglobulin, the antibody was localized on the surface of the schistosomula and at the cell-organism areas of contact. The serum antibody activity appears with the onset of egg production, does not correlate well with the duration of infection, and is relatively specific among schistosome species. Eosinophils and macrophages were demonstrated by electron microscopy to adhere firmly to the surface of the organisms; the latter cells were shown invading the schistosomula. Furthermore, depletion of eosinophils by prior incubation of PEC with monospecific anti-eosinophil serum blocked the early adherence phase, but a substantial part of PEC ability to kill the schistosomula was retained.

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Erythrocyte entry by malarial parasites. A moving junction between erythrocyte and parasite.

Invasion of erythrocytes by merozoites of the monkey malaria, Plasmodium knowlesi, was investigated by electron microscopy. The apical end of the merozoite makes initial contact with the erythrocyte, creating a small depression in the erythrocyte membrane. The area of the erythrocyte membrane to which the merozoite is attached becomes thickened and forms a junction with the plasma membrane of the merozoite. As the merozoite enters the invagination in the erythrocyte surface, the junction, which is in the form of a circumferential zone of attachment between the erythrocyte and merozoite, moves along the confronted membranes to maintain its position at the orifice of the invagination. When entry is completed, the orifice closes behind the parasite in the fashion of an iris diaphragm, and the junction becomes a part of the parasitophorous vacuole. The movement of the junction during invasion is an important component of the mechanism by which the merozoite enters the erythrocyte. The extracellular merozoite is covered with a prominent surface coat. During invasion, this coat appears to be absent from the portion of the merozoite within the erythrocyte invagination, but the density of the surface coat outside the invagination (beyond the junction) is unaltered.

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Uterine leiomyoblastoma: ultrastructural and cytological studies.

A case of uterine leiomyoblastoma is presented with ultrastructural and cytological findings. As in gastric leiomyoblastoma, fine filaments of dimensions consistent with myofilaments and marginal dense bodies are present within the tumor cells, providing evidence for the smooth muscle origin of this neoplasm. Numerous cytoplasmic vacuoles are seen in cellular samples obtained by directly scraping the lesion corresponding to the vacuoles present in glutaraledehyde-fixed material examined by electron microscopy. They do not appear to be artifacts of poor fixation and may be related to the nature of the tumor. Cytologically the cells do not resemble epithelial cells of either squamous or glandular origin and are more consistent with cells of mesenchymal origin.

Adult↗

Purification and properties of alpha-mannosidase from bakers' yeast.

The yeast alpha-mannosidase [EC 3.2.1.24] was purified 1160-fold from the crude extract of the autolysate. The purified preparation was practically free from alpha-glucosidase, beta-glucosidase, alpha-galactosidase, beta-galactosidase, beta-mannosidase, and beta-N-acetylhexosaminidase activities. After the separation of yeast mannan during the purification procedures the enzyme became unstable but could be stored at 5 degrees C for three weeks with 50% loss of activity. The purified enzyme hydrolyzed both aryl and alkyl mannosides, but hydrolysis of yeast mannan proceeded slowly. Yeast mannan and Zn2+ increased the enzyme catalyzed hydrolysis of p-nitrophenyl mannoside, whereas NaN3, monoiodoacetate and methyl alpha-D-mannoside acted as inhibitors. The molecular weight was estimated to be 450,000 by gel filtration.

Glycoside Hydrolases↗

Further evidence for the parasitic origin of the surface coat on Malaria merozoites.

It has been observed that a surface coat, 20 nm in thickness, extends from the plasma membrane of extracellular merozoites and that this coat contains strain-specific antigens. In the present study we determined the possible effect of protein in the culture medium on this structure. The surface coat on merozoites in culture medium devoid of protein was indistinguishable by transmission electron microscopy from the coat in a protein-containing medium. This supports the hypothesis that the surface coat originates from the parasite rather than from adsorbed serum protein.

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Variations in structure and function during the life cycle of malarial parasites.

The fine structure of malarial parasites is reviewed and the function of the intracellular organelles is discussed. When the erythrocytic, exoerythrocytic, and mosquito stages of plasmodia are compared, substantial differences are seen. The major differences involve the amount of surface coat of the motile forms, the structure and function of the mitochondria, and the ingestion and digestion of nutrients. Significant structural differences are also observed between comparable stages of mammalian and avian parasites. These differences indicate that malarial parasites adapt themselves to the different environments in which the parasite resides.When host cell changes induced by malarial parasite infection are reviewed, alterations characteristic of the infecting plasmodia are observed in erythrocytes. Erythrocyte changes include caveola-vesicle complexes, excrescences, and clefts. The caveola-vesicle complexes possess malarial antigens and exhibit pinocytotic activities. The excrescences form focal junctions with adjacent cells and may be responsible for infected erythrocyte sequestration in organs. The significance of these host cell changes specific to certain species of malarial parasite is still unknown.

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Ultrastructure and viability of cryopreserved Plasmodium falciparum.

Cryopreserved chimpanzee erythrocytes infected with Plasmodium falciparum were examined by electron microscopy directly and after a period of culture. Light microscopic observations on the viability of cryopreserved parasites in culture were also made. Parasitaemia data from a chimpanzee infected with parasites cryostored for more than 2(1/2) years are presented. Varying degrees of damage were apparent as the parasites were examined in vitro, for viability, and by electron microscopy. Schizonts appear not to survive the process, whereas the ability of "ring" trophozoite forms to develop into large trophozoites in vitro varies from 20% to 100% in different preparations. Cryopreserved material examined by electron microscope showed degenerative changes in most of the parasites, but after 72 h of cultivation most of the parasites appeared normal. These findings suggest that some organisms may sustain reversible damage during cryostorage.

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Transmission and scanning electron microscopy of host cell entry by Toxoplasma gondii.

Entry by tachyzoites of Toxoplasma gondii, the causative agent of toxoplasmosis, into peritoneal cells was investigated with transmission and scanning electron microscopy. The process of entry is initiated by the parasite contacting the host cell with its anterior end, creating a small depression in the plasmalemma of the host cell. Occasionally, a small portion of the host cell cytoplasm protrudes and contaccts the anterior end of the parasite. A cylindrical structure (35 nm in diameter) extends from the pellicle of the parasite to the host cell. Such structures appear to assist host cell entry by T. gondii. As the entry process progresses, pseudopods of the host cell surround theparasite and finally T gondii becomes intracellular, being located in a vacuole separated from the host cell cytoplasm by a unit membrane. (Am J. Pathol 87:285-296, 1977).

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