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Biomedical subjects

B A Afzelius

Publications and source records attributed to B A Afzelius.

At least 37 records · Page 2Linked to original sources

Ultrastructure of human nasal epithelium during an episode of coronavirus infection.

The nasal epithelium from a young girl was examined by electron microscopy and found to be infected by coronavirus. Virions are seen within and outside the ciliated cells, but not outside or within the goblet cells or other cells of the nasal mucosa. Some virions are located near the microvilli, others in pockets in the apical cell membrane. The cytoplasm contains many small vesicles with a single virion, large apical vesicles containing hundreds of virions, and lysosome-like cytosomes with a moderate number of virions. Some virus-like particles devoid of an electron-dense interior are seen both in the cytosomes and extracellularly. Virus budding was observed in the Golgi apparatus but nowhere else in the cell. The ciliated cells seem not to be destroyed by the viruses, although in many cases the cilia are withdrawn into the cell body. The loss of cilia is likely to cause rhinorrhoea.

Child, Preschool↗

Effects of clofibrate withdrawal on peroxisomes in mouse hepatocytes.

Adult male mice (NMRI strain) were initially treated with 0.5% clofibrate in the diet during four days to obtain a high level of peroxisomes in the hepatocytes. Groups of animals were then given control diet for one, two, three, or four days. Liver samples were cytochemically stained for catalase prior to examination by electron microscopy and subsequent morphometric analysis. Various enzyme activities were assayed in liver homogenates. A sixfold increase in peroxisomal area was found after four days of clofibrate feeding compared with untreated animals. The peroxisomal fractional area was unchanged after one day of refeeding control diet but was reduced with over 50% after two days on control diet and was similar to that in the untreated animals after four days. The size distribution of peroxisomes shifted to smaller values during the breakdown of peroxisomes and was similar to the untreated animals after four days of clofibrate withdrawal. During this process a more heterogeneous staining of peroxisomes was observed. The mitochondrial fractional area was not affected by the treatments, but a shift in size distribution to smaller values was observed in the most induced animals. Protein content was not affected by the treatments. The activity of catalase (twofold induction) decreased progressively to control values during recovery but enoyl-CoA hydratase activity (fourty-fold induction) decreased rapidly. NADPH-cytochrome c reductase activity (twofold induction) decreased rapidly to control levels. Citrate synthase activity was not affected by clofibrate treatment but decreased during recovery. Acid phosphatase activity (repressed) increased to control levels. Cathepsin D activity increased somewhat during recovery while proteolytic activity (towards casein) decreased transiently on control diet.

Acid Phosphatase↗

Section staining for electron microscopy using tannic acid as a mordant: a simple method for visualization of glycogen and collagen.

The possibility that tannic acid and other classical mordants can be used in the electron microscopical section staining technique has been tested. A mordant can be defined as a chemical that combines with both a certain specific tissue component and the staining substance and thereby permits a staining reaction that otherwise will not be obtained. The following features were found to characterize section staining of tannic acid mordanted sections. Tannic acid apparently blocks those sites that normally would be contrasted by uranyl acetate or some other staining compounds. Ribosomes remain unstained. Glycogen particles, on the other hand, were stained, whereas they are not in non-mordanted sections. In fact, glycogen was the only cytoplasmic component to be contrasted by the uranyl acetate, and collagen the only extracellular component. Several different section staining solutions gave the same staining patterns of examined cells and tissues. Specificity of the reaction thus seems to depend on the mordant rather than on the heavy atom section stain. Some other tested mordants, which have also been used in the light microscopical technique, did not give any useful new information.

Animals↗

Ultrastructural analysis of fresh Plasmodium falciparum-infected erythrocytes and their cytoadherence to human leukocytes.

Sixty fresh Plasmodium falciparum isolates obtained from Gambian children with mild or cerebral malaria were investigated by transmission electron microscopy for the expression of knob-like protrusions (K+) on the surface of the infected erythrocytes. More than six-hundred infected erythrocytes were analyzed. Knob-forming parasites were present in all 60 isolates. Although knobless parasites (K-) were found in 25 (42%) of the isolates, only 39 were K-, while 577 were K+. Nine of the 39 K- infected erythrocytes that were studied in greater detail appeared to be asexual parasites because they were either segmented or they lacked mitochondrial DNA-like filaments and cristae, which are abundant in immature gametocytes. No difference was observed in the relative frequency of K+K- infected erythrocytes in isolates from patients with mild or cerebral malaria. Binding of both knobby and knobless infected erythrocytes to autologous leukocytes including monocytes, neutrophils, lymphocytes and plasma cells was found in some of the primary in vitro cultures. By using P. falciparum laboratory strains of known phenotypes and leukocytes from healthy blood bank donors, it was established that this novel adherence phenomenon was related to that of cytoadherence to certain melanoma or endothelial cells. Cytoadherent infected erythrocytes that bind to leukocytes enhance antibody-independent phagocytosis and induce cellular aggregation, while non-cytoadherent or rosetting infected erythrocytes do not.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cytoadherence of knobby and knobless Plasmodium falciparum-infected erythrocytes.

Cytoadherence of Plasmodium falciparum-infected erythrocytes to melanoma cells was analysed using strains or isolates of parasites expressing or not expressing knobs (K+ or K- phenotype) on the erythrocyte surface. Both K+ and K- parasites had the capacity to cytoadhere to melanoma cells. Using a panel of melanoma cell lines with different surface expression of the cytoadherence receptors CD36, thrombospondin and ICAM-1 indicated that CD36 was the major receptor for parasites of both K+ and K- phenotypes. Binding competition experiments between K+ and K- -infected erythrocytes suggested that K+ cytoadherence is of higher affinity than that of K- parasites. However, some K- cytoadherence was also found in isolates containing mixed populations of K+ and K- parasites. The interaction of the two types of infected erythrocytes with melanoma cells also differed ultrastructurally, erythrocytes of K+ phenotype showing intimate interdigitations with microvilli on the melanoma cells, while erythrocytes of K- phenotype displayed more separated interactions with fewer sites of contact and involving only a few melanoma cell microvilli. One and the same infected erythrocyte may co-express the ligand for CD36-mediated cytoadherence and the structures mediating binding of uninfected erythrocytes to form rosettes.

Animals↗

A strategy for the reconstruction of structures possessing axial symmetry: sectioned axonemes in sperm flagella.

The images of complex biological structures seen in the electron microscope, possessing an n-fold rotational symmetry, can be enhanced by averaging the axially repeating motif in order to improve their signal-to-noise ratio; this requires that the slices with n-fold symmetry do not exhibit distortions relative to one another. A strategy is proposed to detect the relative distortions and to remove them in order to obtain reliable results from the averaging process. Extensive use is made of cross-correlation analysis and of interpolation; because the procedure involves iterated resampling of the image, it is essential to adopt interpolation algorithms which preserve the spectral power. The procedure is illustrated by the analysis of transverse sections of the sperm flagellum axoneme of a stick insect; it might be used for the reconstruction of microtubules, nuclear pore complexes, virus capsids, and other supramolecular aggregates. The reconstructed images of axoneme sections reveal new information about the interactions between adjacent doublets; in particular, a double conjunction connecting the inner dynein arm with the nearest B-tubule has been consistently observed.

Animals↗

The spermatozoon of the Chinese bitterling, Rhodeus sericeus sinensis (Cyprinidae, Teleostei).

The bitterling spermatozoon has been examined by electron microscopy using sectioned material and freeze-fracture replicas. Its structure is characterized by a higher degree of order than is seen in spermatozoa of related teleosts. The intramembrane particles in the cell membrane are arranged in a large (around 1 micron in diameter) hexagonal grating pattern at a certain site of the sperm head. The flagellum protrudes from a site slightly posteriorly to this hexagonal grating and is surrounded by a cytoplasmic sleeve that contains regularly spaced filaments in a helical arrangement. The sperm midpiece contains a large mitochondrion enclosed in an elaborate system of membranous cisternae and narrow tubules. Fertilization in the bitterling takes place in the gill chamber of freshwater mussels--a unique brood chamber and site of fertilization for a vertebrate.

Animals↗

Time-dependent effects of the hypolipidemic agent clofibrate on peroxisomes and mitochondria in mouse hepatocytes.

Mice of the NMRI strain were fed a diet containing 0.5% clofibrate up to 25 days to study its effect on peroxisomes and mitochondria in the hepatocytes. Liver samples were analyzed by electron microscopy after cytochemical staining for catalase using diaminobenzidine in order to detect all peroxisomes for a quantitative analysis. The liver weights increased significantly after two days of treatment and reached a maximal level of 40% over the control animals. The number of peroxisomal profiles increased 3- to 4-fold at maximal proliferation and their average area nearly doubled, hence the number of peroxisomes with large diameters was increased making a 5-fold increase within the total cytoplasmic area. The size distribution of the peroxisomes was changed at day two of treatment to a more spread and even distribution going up in a higher size range. The mitochondrial profiles were less affected but there was a slight increase in their number, a small decrease in average area, a transient decrease in axial ratio but no significant effects on their cytoplasmic fractional area. The frequency distribution of mitochondrial area was shifted somewhat to lower values by clofibrate. At the end of the treatment the hepatocytes contained about the same number of peroxisomal and mitochondrial profiles but the latter were still occupying a two-fold higher fractional volume of the cytoplasm. Mouse hepatocytes thus seem to be a good system for studies on peroxisomal biogenesis as is their counterpart in rats, although they differ in their levels of various peroxisomal enzymes.

Animals↗

Ciliary orientation in the "immotile cilia" syndrome.

Ciliary orientation was studied in 43 patients with the "immotile cilia" syndrome. Twenty-four of these patients had total situs inversus. One mucosal specimen was taken from uterine cervical epithelium, 2 were from bronchial mucosa and 40 from nasal mucosa. The orientation of the cilia was measured from micrographs using a semiautomatic image analyzer (IBAS I). The results from patients were compared with those of 10 control subjects. The mean standard deviation and its standard deviation of the angles of ciliary orientation was 39.7 degrees +/- 9.2 degrees in 43 patients and 27.4 degrees +/- 4.3 degrees in the control group. The difference between the groups is highly significant statistically (P less than or equal to 0.001). However, there were no statistically significant differences in the standard deviations of ciliary orientation between the fields sectioned near the cell membrane or near the ciliary tip. We were also unable to find any significant differences in the standard deviations of the ciliary angles in the specimens taken from brush biopsies and excisional biopsies. There were also no statistically significant differences between the standard deviations of the ciliary angles for the groups with or without situs inversus. If 35 degrees is considered to be the limit value for the mean standard deviation between normal and pathological specimens in our total material, this would give a specificity of 0.90 and a sensitivity of 0.72.

Biopsy↗

Ultrastructural patterns of the flagellar axoneme in the non-motile part of the mole-cricket sperm.

The mole-cricket spermatozoon (Gryllotalpa gryllotalpa) has a motile anterior tail region and an immotile posterior region. The posterior portion appears stiff and its microtubular doublets and central singlet microtubules are swollen, apparently due to an excess of material within them. In particular, doublet number 6 is of an unusually large size. The general organization of the axoneme is also modified by a loss of dynein arms and spokes in the posterior portion. When studied by a fixation technique that involves tannic acid to outline the protein molecules and PA-TCH-Ag method for staining polysaccharides it could be seen that the microtubular doublets and accessory microtubules contain rounded globules surrounded by polysaccharides. The arrangement of protofilaments within the microtubular walls is visible both in the anterior tail region with normal doublets and in the posterior region with degenerated doublets.

Animals↗

Microtubules and their protofilaments in the flagellum of an insect spermatozoon.

Spermatozoa of stick insects have nine accessory tubules, which surround the nine outer microtubular doublets and the two inner microtubular singlets. When fixed in a fixative that was designed to minimize protein denaturation (glutaraldehyde and tannic acid, no osmium post-fixation but block staining with uranyl acetate in water) the accessory tubules were seen to contain 17 protofilaments of the same type as those in the 9 + 2 microtubular doublets and singlets. The protofilaments in accessory tubules and other microtubules were roughly triangular. When studied by Markham's photographic method a somewhat different tilt of the two longer sides was seen; this makes it possible to distinguish a polarity in the microtubules, i.e. to differentiate between a microtubule that is viewed from its (-)end to its (+)end from one that is viewed in the opposite direction. The dynein arms of the doublets can be used as an independent type of marker for the polarity. In a computer-aided analysis of the fine structure of the tail axoneme, the A-subtubules of the outer doublets were seen to be not quite equidistant; rather, there were somewhat widened electron-dense interspaces in the ring of protofilaments in four places. The locations of these widened interspaces coincide with the attachment sites for the spoke, the inner dynein arm, the outer dynein arm, and the intertubular material. It is tentatively concluded that proteins of these structures, and perhaps also other microtubule-associated proteins, may be anchored deep within the wall of a microtubule rather than just superficially along it.

Animals↗

Endospore-forming filamentous bacteria symbiotic in termites: ultrastructure and growth in culture of Arthromitus.

Many morphologically distinguishable filamentous spore-forming bacteria symbiotic in the paunch (hypertrophied hindguts) of wood-eating insects have been seen since Arthromitus was first described and named as a plant by Leidy in 1850. Previous descriptions were inadequate for acceptance of the group in modern bacteriological literature. Twenty-two distinguishable arthromitids in nine different arthropod hosts are recorded on the basis of microscopic studies. Five are named, including two whose ultrastructure are detailed: Arthromitus chasei sp. nov. that lives in the damp wood-eating termite Zootermopsis angusticollis (from the west coast of North America) and Arthromitus reticulitermitidis sp. nov. from the subterranean west coast termite Reticulitermes tibialis. A pterotermiditis from the desert termite Pterotermitidis occidentis; A. zootermopsidis, also from Z. angusticollis; and A. cristatus (Leidy, 1881) from Reticulitermes flavipes of eastern North America are also named here. Characterized by trichomes that show a morphogenetic sequence from no spores through immature spores to mature spores with spore filaments, Arthromitus symbionts can be identified as members of the genus by light microscopy and habitat. Electron microscopy reveals their remarkable complexity. They attach by spore filaments to various objects including the host gut wall; their maturation extends distally toward the termite lumen. By surface sterilization of the termite, maceration of the paunch, exposure to boiling temperatures and plating on soft acetate agar, the heat resistant nature of the spores and facultatively aerobic nature of Arthromitus sp. (from Zootermopsis) was demonstrated.

Animals↗

Ontogenetical changes in adipose tissue of the cat: convertible adipose tissue.

The ultrastructural characteristics of the inguinal, interscapular, and perirenal adipose tissue in kittens and cats were studied. There were no qualitative differences among adipocytes in the three anatomical areas. The only recorded difference was in the amount of lipids stored in the adipocytes in younger stages. Immediately after birth lipids occupied 25% of the volume in the inguinal area, 15% in interscapular fat tissue, and 10% in perirenal fat tissue. At this stage the adipose tissue morphologically resembled brown adipose tissue (BAT) of rodents. Two weeks after birth, lipids accumulated and adipocytes in the inguinal area became unilocular and appeared similar to white adipose tissue (WAT). A similar transition occurred approx 25 days after birth in interscapular fat and approx 6 weeks after birth in the perirenal area. No morphological signs of any cell degradation or destruction, nor any increased activity of preadipocytes, were seen during this conversion from BAT-like to WAT-like adipose tissue. The conversion of the adipose tissue was correlated with a decrease in vascularization and innervation, a loss of intercellular connections, and a changed mitochondrial population. Mitochondria in multilocular adipocytes resembled those in typical BAT which contain uncoupling protein ("UC-mitochondria"). After conversion to unilocular adipocytes the amount of mitochondria was halved, their cristae even more reduced, and their appearance was of a WAT-type (UCP-lacking mitochondria, which are coupled under physiological conditions; "C-mitochondria"). Since this category of adipose tissue differs from both typical brown and white adipose tissue, the name "convertible adipose tissue" (CAT) is proposed. Apparently adipose tissue from comparatively large mammals is of this convertible type.

Adipose Tissue↗

Effects of clofibrate treatment and of starvation on peroxisomes, mitochondria, and lipid droplets in mouse hepatocytes: a morphometric study.

Adult male mice of the NMRI strain were treated with a diet containing 0.5% clofibrate for 4 days to study its effects on peroxisomes, mitochondria, and lipid droplets in hepatocytes. Animals were also starved overnight to study the additional effects of starvation. Starvation of control animals had small effects on peroxisomes while the mitochondria became enlarged and occupied more of the cytoplasm. The number and fractional area of lipid droplets increased fivefold. Clofibrate treatment caused a doubling in number and average size of peroxisomes. No significant effects were observed in the number of mitochondrial profiles or lipid droplets although the size of the latter decreased to a third the value of the fed control. Starvation of clofibrate-treated animals led to a slight increase in the number of peroxisomes although their average size decreased by 30%. Mitochondrial average area increased and their fractional cytoplasmic area increased despite the decrease in numerical density. The number of lipid droplets increased twofold compared to that of clofibrate-treated animals while the size was not affected.

Animals↗

Ultrastructure and function of thrombocytes in a family with congenital bleeding tendency.

Freshly isolated blood from controls and from three members of a family with congenital bleeding tendency were examined with respect to ultrastructure of platelets. The fixation protocol, which included tannic acid, permitted the visualization of substructures in the marginal bundle. Whereas nearly all platelets from the controls had a discoid shape, those from one of the patients (mother of the other two cases) were mostly flat or else roundish with two or three marginal bundles in different orientations. One portion of the thrombocyte fraction of each person was briefly exposed to adenosine diphosphate in order to activate the platelets and thereby to induce shape changes. Most platelets from the controls reacted as expected by rounding up and by projecting some pseudopods. A certain percentage (between 10 and 20%) of the microtubules in these activated platelets had 14 or even 15 protofilaments. The platelets from patients (and in particular the mother) either reacted by an abnormal bulging of the cytoplasm or did not react visibly at all. The microtubules retained their 13-subunit composition or in the abnormally reacted ones often had an open C-shaped cross-sectional profile. The bleeding tendency in this family might be due to a defect in the cytoskeleton of the platelet making it unable to react properly to stimuli.

Adenosine Diphosphate↗

The substructure of marginal bundles in human blood platelets.

A fixation protocol was worked out which permitted the visualization of the substructure of microtubules in human blood platelets. This protocol included a brief prefixation with 0.1% glutaraldehyde in a "polymerization mix buffer" including GTP, followed by a fixation mixture containing glutaraldehyde and tannic acid, and postfixation in 0.1% osmium tetroxide. The 13 individual protofilaments of the microtubules were clearly visible and had a triangular cross-sectional area. Filaments could be seen running alongside the microtubules and connected to them by many links. We term these filaments the "intertubular filaments." Some microtubules were incomplete and had a C-shaped profile; by serial sectioning it was seen that such microtubules were close to the terminal ends of the microtubules. A low percentage of the platelets had two marginal bundles oriented perpendicularly to each other.

Blood Platelets↗

Early changes in the substructure of the marginal bundle in human blood platelets responding to adenosine diphosphate.

Human blood platelets were exposed to 10 microM adenosine diphosphate in order to induce shape changes and their ultrastructure was examined in preparations fixed with a glutaraldehyde-tannic acid mixture. Within 5 sec after the addition of ADP the marginal bundle along the perimeter of the platelet had undergone profound changes. The intertubular filaments of the marginal bundle were lost and, probably as a consequence, the microtubules were dispersed. The microtubules also had fragmented and appeared as straight rods rather than smoothly curved coils. Profiles with 14 or 15 protofilaments rather than the common 13-unit pattern were seen in 16% of the cross-sectioned microtubules. The proportion of incomplete (C-shaped) microtubules or of microtubules with an electron-dense interior also had increased and were interpreted as sections near the terminal ends of a microtubule. We interpret the C-shaped and the "filled" microtubules as signs of a disassembled microtubule and the 14- and 15-protofilament profiles as probably representing new (ends of) microtubules reassembled with a defective control of the proper structure.

Adenosine Diphosphate↗

Microtubules in the spermatids of stick insects.

Spermatids from two phasmid species were seen to possess an unusually large amount of microtubules along the nucleus and tail. Some of the microtubules have a loosely fitting sleeve for half a micron or more. During late stages in spermiogenesis the microtubules aggregate and form one or several "microtubular crystals" consisting of electron-lucid tubular elements with a diameter of about 360 A. The tail flagellum contains five kinds of microtubular structures, which all have a substructure of longitudinal protofilaments that is clearly visible after fixation in the presence of tannic acid. The so-called accessory tubules have 17 protofilaments that have the same appearance as that in ordinary, 13-unit microtubules, but are somewhat thicker than those. It is evident that the protofilaments in both the 17-unit and the 13-unit microtubules run parallel or nearly parallel to the long axis of the microtubules. It is of interest that both types of microtubules possess a prime number of protofilaments which may give the fagellum certain functional advantages.

Animals↗