Membrane vesicles in magnetotactic bacteria

Magnetotactic bacteria are microorganisms that respond to magnetic fields. We have studied the surface ultrastructure of Magnetospirillum magnetotacticum and uncultured magnetotactic bacteria from a marine environment using transmission electron microscopy and freeze-etching. Numerous membrane vesicles were observed on the surface of Magnetospirillum magnetotacticum bacteria. All uncultured magnetotactic bacteria presented membrane vesicles on their surface in addition to an extensive capsular material and an S-layer formed by particles arranged in a hexagonal symmetry. We did not observe any indication of electron-dense precipitation on the surface of these microorganisms. Our results indicate that membrane vesicles are a common characteristic of magneto-tactic bacteria in natural sediments.     

Adhesive cell cultivation on polymer particle having grafted epoxy polymer chain

In this study, we synthesized a new cell immobilization support having poly(glycidyl methacrylate) as a graft polymer chain and used this support for cell cultivation. Base polymer particle was synthesized by suspension polymerization and epoxy polymer chain was extended from particle surface on graft polymerization. Produced polymer particles had broad particle size distribution ranging from 20 to 1000 μm and the degree of polymerization of grafted polymer chain was ranged from 500 to 1000. The effects of various factors, such as grafted polymer chain length and its surface density, composition of base polymer network and graft polymer chain, on the cell growth of murine fibroblast cell line (MS-5 cell) on polymer particle were studied. This polymer particle could cultivate not only fibroblast cell line but also epidermal cell line (HeLa cell), osteoblast cell line (MC3T3E1 cell), and chondrocyte cell line (ch-8 cell) on its surface. Growth rate is almost the same as that of cells using poly(styrene) tissue culture dish. To apply this cell cultivation system for examination of cell co-culture, HeLa cell immobilized on 100 μm of polymer particle was successfully co-cultured with MS-5 cell immobilized on 300 μm of polymer particle for four weeks.     

Analysis of the murine sperm surface with monoclonal antibodies

Hybridomas secreting monoclonal antibodies reactive with murine spermatozoa were produced by fusion of myeloma cells with spleen cells from C57BL/6J mice immunized with spermatozoa from mice of the same strain. All antisperm antibodies were of the mu (mu) immunoglobulin heavy chain class; only one (MS-1) bound S. aureus protein A. Antibody MS-1 recognized an antigen present on the sperm acrosomal cap, on the surface of cells from liver and kidney and from some cultured cell lines. The subunit molecular weight (69000) of the polypeptide reactive with MS-1 was determined by SDS-PAGE analysis of sperm membrane proteins followed by their electrophoretic transfer to nitrocellulose.     

Diversity and vertical distribution of magnetotactic bacteria along chemical gradients in freshwater microcosms

The vertical distribution of magnetotactic bacteria along various physico-chemical gradients in freshwater microcosms was analyzed by a combined approach of viable cell counts, 16S rRNA gene analysis, microsensor profiling and biogeochemical methods. The occurrence of magnetotactic bacteria was restricted to a narrow sediment layer overlapping or closely below the maximum oxygen and nitrate penetration depth. Different species showed different preferences within vertical gradients, but the largest proportion (63-98%) of magnetotactic bacteria was detected within the suboxic zone. In one microcosm the community of magnetotactic bacteria was dominated by one species of a coccoid "Alphaproteobacterium", as detected by denaturing gradient gel electrophoresis in sediment horizons from 1 to 10 mm depth. Maximum numbers of magnetotactic bacteria were up to 1.5 x 10(7) cells/cm3, which corresponded to 1% of the total cell number in the upper sediment layer. The occurrence of magnetotactic bacteria coincided with the availability of significant amounts (6-60 microM) of soluble Fe(II), and in one sample with hydrogen sulfide (up to 40 microM). Although various trends were clearly observed, a strict correlation between the distribution of magnetotactic bacteria and individual geochemical parameters was absent. This is discussed in terms of metabolic adaptation of various strains of magnetotactic bacteria to stratified sediments and diversity of the magnetotactic bacterial communities.     

The Structure of Mycoplasma pneumoniae as Determined by the Freeze-Substitution Technique

The ultrastructure of Mycoplasma pneumoniae FH was examined by a mild fixation method, the freeze-substitution technique, for thin-section transmission electron microscopy and the following new findings were obtained. In the cytoplasm, no nuclear region could be clearly identified. The cytoplasm was filled with many ribosome-like particles, fine fibers and electron-dense particles. The electron-dense particles appeared to be similar to the particles found in the nucleoid region of Pseudomonas aeruginosa and might therefore possibly be a kind of DNA binding protein. The cell surface was completely enveloped with a thin opaque layer. The presence of this surface layer prevented any direct contact of the cell surface with that of the two M. pneumoniae cells.     

Hemagglutination Activity and Localization of Fc Receptor of Group A and G Streptococci

The IgG-Fc binding activity and binding sites on the cell surface of streptococci, strains AR1 (group A) and G148 (group G), and Staphylococcus aureus strain Cowan I were examined by hemagglutination (HA) and immunoelectron microscopic methods. No distinct difference was observed in the HA activity among these three strains. However, the strains differed in the distribution of Fc receptor. Cowan I cells (having protein A) were heavily covered with two layers of ferritin particles, whereas AR1 cells were heavily covered with a single, rough layer of ferritin particles. G148 cells (having protein G) were labeled with a relatively thin, rough ferritin layer. The trypsin susceptibility of the Fc receptors of the AR1 strain was much higher than that of the G148 strain. These results suggest that both streptococcal strains are distinctly different in the arrangement or in the conformation of the Fc receptor from the Cowan I strain. It is also suggested that the Fc receptor molecules of the streptococcal strains differ from each other not only in conformation but also in trypsin susceptibility.     

Envelope ultrastructure of uncultured naturally occurring magnetotactic cocci

Magnetotactic bacteria are microorganisms that respond to magnetic fields. We studied the surface ultrastructure of uncultured magnetotactic cocci collected from a marine environment by transmission electron microscopy using freeze-fracture and freeze-etching. All bacteria revealed a Gram-negative cell wall. Many bacteria possessed extensive capsular material and a S-layer formed by particles arranged with hexagonal symmetry. No indication of a metal precipitation on the surface of these microorganisms was observed. Numerous membrane vesicles were observed on the surface of the bacteria. Flagella were organized in bundles originated in a depression on the surface of the cells. Occasionally, a close association of the flagella with the magnetosomes that remained attached to the replica was observed. Capsules and S-layers are common structures in magnetotactic cocci from natural sediments and may be involved in inhibition of metal precipitation on the cell surface or indirectly influence magnetotaxis.     

磁泳分离细菌新方法的研究

从酸性矿坑水中富集培养分离到的嗜酸氧化亚铁硫杆菌（Acidithiobacillus ferrooxidans,A.ferrooxidans）[1-2] 菌同趋磁细菌具有一定的相似性。通过显微镜观察发现,部分浸矿细菌在外加磁场的作用下具有微弱的趋磁性,基于菌种的这种特性,设计了磁泳分离仪,对其在磁场作用下泳动（磁泳）进行分析,经磁泳后的近磁、远磁菌的生理特性有较大的差异。从用涂布平板法获得的近磁菌纯培养A. ferrooxidans菌体中,分离得到纳米磁性颗粒,能谱分析表明,其主要成分为Fe和O元素。实验结果证明,A. ferrooxidans具有微弱趋磁性,采用磁泳分离该类菌体内含有磁性颗粒的细菌是可行的,这一分离技术的进一步完善和改进将为传统的微生物菌种分离提供一种新型分离技术,也将大大促进趋磁细菌的研究,而且它与浸矿工艺的结合将大大促进我国生物冶金的研究步伐。     

Ultrastructure of the Surfaces of Cells Infected with Avian Leukosis-Sarcoma Viruses

When stained with ruthenium red (RR), chick embryo cells infected with various strains of Rous sarcoma virus (RSV) and with avian leukosis viruses RAV-1 and RAV-3 showed an increase in the layer of acid mucopolysaccharides (AMPS) at their surfaces as compared with uninfected cells. This increase was most prominent in cells infected with the Fujinami strain of RSV. The layer was resistant to digestion with neuraminidase or trypsin but was readily removed by exposure to hyaluronidase. The thickness of this AMPS layer was not correlated with the varying degree of loss of contact inhibition exhibited by cells infected with the different strains of virus. The staining of the cell envelope with a solution of phosphotungstic and chromic acids (PTA-CR) suggested the presence of glycoproteins. The outer surface of the virions showed the same staining as the cell surface with RR and PTA-CR, and the budding virus particle was seen to incorporate the RR layer of the cell into its structure. The RR layers of cells and virions appeared to fuse, as did those between virus particles, suggesting that these layers play a role in the aggregation of virus particles and in their adherence to the surface of the cell.     

Characterization of Bacterial Magnetotactic Behaviors by Using a Magnetospectrophotometry Assay

Magnetotactic bacteria have the unique capacity of synthesizing intracellular single-domain magnetic particles called magnetosomes. The magnetosomes are usually organized in a chain that allows the bacteria to align and swim along geomagnetic field lines, a behavior called magnetotaxis. Two mechanisms of magnetotaxis have been described. Axial magnetotactic cells swim in both directions along magnetic field lines. In contrast, polar magnetotactic cells swim either parallel to the geomagnetic field lines toward the North Pole (north seeking) or antiparallel toward the South Pole (south seeking). In this study, we used a magnetospectrophotometry (MSP) assay to characterize both the axial magnetotaxis of “Magnetospirillum magneticum” strain AMB-1 and the polar magnetotaxis of magneto-ovoid strain MO-1. Two pairs of Helmholtz coils were mounted onto the cuvette holder of a common laboratory spectrophotometer to generate two mutually perpendicular homogeneous magnetic fields parallel or perpendicular to the light beam. The application of magnetic fields allowed measurements of the change in light scattering resulting from cell alignment in a magnetic field or in absorbance due to bacteria swimming across the light beam. Our results showed that MSP is a powerful tool for the determination of bacterial magnetism and the analysis of alignment and swimming of magnetotactic bacteria in magnetic fields. Moreover, this assay allowed us to characterize south-seeking derivatives and non-magnetosome-bearing strains obtained from north-seeking MO-1 cultures. Our results suggest that oxygen is a determinant factor that controls magnetotactic behavior.Magnetotactic bacteria are morphologically, metabolically, and phylogenetically diverse prokaryotes (1, 11). They synthesize unique intracellular organelles, the magnetosomes, which are single-domain magnetic crystals of the mineral magnetite or greigite enveloped by membranes. Magnetosomes are usually organized in a chain(s) within the cell and cause the cell to align along geomagnetic field lines while it swims. The highest numbers of magnetotactic bacteria are generally found at, or just below, the oxic-anoxic transition zone (OATZ) or redoxocline in aquatic habitats (1). Early studies showed that Northern Hemisphere magnetotactic bacteria swim preferentially northward in parallel with the geomagnetic field lines (north seeking [NS]) (2) and that those from the Southern Hemisphere swim preferentially antiparallel to the geomagnetic field lines to the magnetic South Pole (south seeking [SS]) (4). The geomagnetic field is inclined downward from horizontal in the Northern Hemisphere and upward in the Southern Hemisphere, with the inclination magnitude increasing from the equator to the poles. Therefore, magnetotaxis might guide cells in each hemisphere downward to less-oxygenated regions of aquatic habitats, where they would presumably stop swimming until conditions change (1). A recent study reported the coexistence of both NS and SS magnetotactic bacteria in the Northern Hemisphere, which conflicts with the prevalent model of the adaptive value of magnetotaxis (14).Under laboratory conditions, magnetotactic bacteria form microaerophilic bands of cells in oxygen-gradient medium. In fact, magnetotaxis and aerotaxis work together in these bacteria, and the behavior observed has been referred to as “magnetoaerotaxis.” Two different magnetoaerotactic mechanisms, termed polar and axial, are found in different bacterial species (6). The magnetotactic bacteria, principally the magnetotactic cocci, that swim persistently in one direction along the magnetic field (NS or SS) are polar magnetoaerotactic. Magnetotactic bacteria, especially the freshwater spirilla, that swim in either direction along the magnetic field lines with frequent, spontaneous reversals of swimming direction without turning around are axial magnetoaerotactic. For polar magnetotactic bacteria, the magnetic field provides an axis and a direction for motility, whereas for axial magnetotactic bacteria, the magnetic field provides only an axis of motility. The two mechanisms can best be seen in flattened capillary tubes containing suspensions of cells in reduced medium in a magnetic field oriented parallel to the capillary. An oxygen gradient forms along the tube, beginning at the ends of the capillary, with one oriented parallel and the other antiparallel to the magnetic field (1). Band formation by axial magnetoaerotactic cells, such as Magnetospirillum magnetotacticum cells, occurs at both ends of the capillary. Rotation of the magnetic field by 180° after the formation of the bands causes the cells in both bands to rotate 180°, but the bands remain intact. In contrast, band formation by polar magnetoaerotactic cells, such as the marine cocci, occurs only at the end of the capillary for which the magnetic field and the oxygen concentration gradient are oriented opposite to each other. Rotation of the magnetic field by 180° after the formation of the band causes the cells in the band to rotate 180° and swim away, resulting in the dispersal of the band (1). In this study, we developed a magnetospectrophotometry (MSP) assay that provides an alternative method for the quantitative and versatile characterization of the two magnetotactic mechanisms. Using this assay, we demonstrated the effect of artificial magnetic fields on the generation of homogeneous NS or SS magnetotactic bacterial populations.     

Iron reduction byAquaspirillum magnetotacticum

Iron reductase activity in cell extracts ofAquaspirillum magnetotacticum strain MS-1 (wild type) or nonmagnetotactic mutant strain NM-1A was located primarily in the periplasm. Cytoplasm contained 20%–35% and membrane fractions 3% of total iron reductase activity detected. Iron reduction was reversibly inhibited by oxygen, required -NADH, and was enhanced by flavins. Reduced disulfide bonds and uncomplexed sulfhydryl groups were necessary for reductase activity. Respiratory inhibitors did not appreciably affect iron reductase activity. Iron complexed with quinic acid, dihydroxybenzoic acid, acetohydroxamic acid, citric acid, or deferrioxamine B was reduced by soluble iron reductases of strain MS-1.     

Ultrastructural study of polymyxin-resistant isolates of Pseudomonas aeruginosa.

Upon exposure to 6,000 U of polymyxin B sulfate per ml, cells of the polymyxin-sensitive PAO 1 strain of Pseudomonas aeruginosa displayed in thin sections long projections arising from the outer membrane of the cell wall and extensive cytoplasmic degradation with accumulation of cytoplasmic membrane infoldings. Polymyxin-resistant isolates derived from the PAO 1 strain, however, grew well in the presence of 6,000 U of polymyxin per ml and exhibited none of these effects, having instead the appearance of a typically healthy cell. Freeze-etching of cells of the sensitive strain grown in basal medium without polymyxin revealed a concave cell wall layer studded with numerous particles. Freeze-etching of cells of the resistant isolates grown in basal medium containing 6,000 U of polymyxin per ml revealed a concave cell wall layer (i.e., the outer half of the outer membrane) in which most of these particles were absent. Thus, acquisition of resistance to polymyxin was correlated with an alteration in the architecture of the outer membrane. When the resistant isolates were grown in the basal medium lacking polymyxin and then freeze-etched, the particle distribution in the concave cell wall layer resembled that of the sensitive parent strain. The cells had regained sensitivity to polymyxin upon suspension in medium containing 6,000 U/ml as determined by their failure to grow and by internal damages seen in thin sections. These cells also had acquired increased sensitivity to ethylenediaminetetraacetate, whereas the polymyxin-resistant cells grown in the presence of polymyxin were resistant to lysis by ethylenediaminetetraacetate. The polymyxin-resistant isolates were not stable mutants but instead represented an adaptive response to the presence of polymyxin in the medium.     

Preparation of Genomic DNA from a Single Species of Uncultured Magnetotactic Bacterium by Multiple-Displacement Amplification

Magnetotactic bacteria comprise a phylogenetically diverse group that is capable of synthesizing intracellular magnetic particles. Although various morphotypes of magnetotactic bacteria have been observed in the environment, bacterial strains available in pure culture are currently limited to a few genera due to difficulties in their enrichment and cultivation. In order to obtain genetic information from uncultured magnetotactic bacteria, a genome preparation method that involves magnetic separation of cells, flow cytometry, and multiple displacement amplification (MDA) using φ29 polymerase was used in this study. The conditions for the MDA reaction using samples containing 1 to 100 cells were evaluated using a pure-culture magnetotactic bacterium, “Magnetospirillum magneticum AMB-1,” whose complete genome sequence is available. Uniform gene amplification was confirmed by quantitative PCR (Q-PCR) when 100 cells were used as a template. This method was then applied for genome preparation of uncultured magnetotactic bacteria from complex bacterial communities in an aquatic environment. A sample containing 100 cells of the uncultured magnetotactic coccus was prepared by magnetic cell separation and flow cytometry and used as an MDA template. 16S rRNA sequence analysis of the MDA product from these 100 cells revealed that the amplified genomic DNA was from a single species of magnetotactic bacterium that was phylogenetically affiliated with magnetotactic cocci in the Alphaproteobacteria. The combined use of magnetic separation, flow cytometry, and MDA provides a new strategy to access individual genetic information from magnetotactic bacteria in environmental samples.Magnetotactic bacteria synthesize nanosized intracellular magnetic particles, also referred to as magnetosomes, by accumulating iron ions from the environment. Since the first report on the identification of magnetotactic bacteria (2), the morphological and phylogenetic diversity of these organisms has been observed in various aquatic environments (12, 25, 27, 30). However, bacterial strains available in pure culture are currently limited to a few genera. Desulfovibrio magneticus strain RS-1 is the only isolate of magnetotactic bacteria that is classified among the Deltaproteobacteria (13, 23), while Magnetospirillum spp., marine magnetic vibrio strain MV-1, and “Magnetococcus strain MC-1” are phylogenetically affiliated within the Alphaproteobacteria group (24, 27). This limitation is mainly because not much is known about their metabolic requirements, culturing conditions, and obligate coculture requirements.Isolation and enrichment of magnetotactic bacteria are generally conducted by applying a magnetic field to a container containing a sediment sample from the environment. The capillary racetrack method is a highly selective enrichment technique that separates magnetotactic bacteria from other contaminants (31). The magnetic separation method that involves the use of a large glass apparatus is efficient and suitable for analyzing samples containing more than 100 ml of sediment and water (12, 16). These techniques have been applied to investigate community structure and phylogenetic diversity of uncultured magnetotactic bacteria in the environment based on 16S rRNA analyses (3, 7, 26, 29). In a recent study, DNA isolation enabling gene cloning was examined by magnetically collecting a large number of magnetotactic cells from environmental samples, and two gene fragments, probably containing parts of magnetosome islands (MAIs) derived from magnetotactic bacteria of the Alphaproteobacteria, were identified (12). However, this approach allows only for sequence gene information to be obtained from a heterogeneous bacterial community in the sample.Multiple displacement amplification (MDA) can generate microgram quantities of high-quality DNA sample from a few femtograms of DNA template (5, 6). We previously revealed that MDA is a powerful tool for whole-genome amplification from the metagenome of an uncultured bacterial community (32). Studies have been conducted to determine the efficacy of MDA for analyzing genomic DNA preparations from a limited number of bacterial cells (14, 17, 21, 22, 28). Complete genomic sequencing of an uncultured gut symbiont in termites has been achieved using MDA products amplified from approximately 1,000 cells (9). Partial genome sequencing using MDA products from a single uncultured cell has also been reported (17, 22). Such targeted genome analyses using MDA products from a single cell or genetically identical microorganisms is advantageous because it allows the assignment of individual genes to the corresponding microorganisms.In this study, an improved genome preparation method involving racetrack purification and flow cytometry followed by MDA was investigated by using a small number of uncultured magnetotactic bacteria. This method can be used for the identification of new genes from rare magnetotactic bacteria in environmental samples.     

Characterization of a spontaneous nonmagnetic mutant of Magnetospirillum gryphiswaldense reveals a large deletion comprising a putative magnetosome island

Frequent spontaneous loss of the magnetic phenotype was observed in stationary-phase cultures of the magnetotactic bacterium Magnetospirillum gryphiswaldense MSR-1. A nonmagnetic mutant, designated strain MSR-1B, was isolated and characterized. The mutant lacked any structures resembling magnetosome crystals as well as internal membrane vesicles. The growth of strain MSR-1B was impaired under all growth conditions tested, and the uptake and accumulation of iron were drastically reduced under iron-replete conditions. A large chromosomal deletion of approximately 80 kb was identified in strain MSR-1B, which comprised both the entire mamAB and mamDC clusters as well as further putative operons encoding a number of magnetosome-associated proteins. A bacterial artificial chromosome clone partially covering the deleted region was isolated from the genomic library of wild-type M. gryphiswaldense. Sequence analysis of this fragment revealed that all previously identified mam genes were closely linked with genes encoding other magnetosome-associated proteins within less than 35 kb. In addition, this region was remarkably rich in insertion elements and harbored a considerable number of unknown gene families which appeared to be specific for magnetotactic bacteria. Overall, these findings suggest the existence of a putative large magnetosome island in M. gryphiswaldense and other magnetotactic bacteria.     

William Trager: An Appreciation

SYNOPSIS. Additional information on host interactions with trypanosomatid membranes was obtained from studies of a monomorphic strain of Trypanosoma brucei harvested at peak parasitemia from intact and lethally irradiated rats. Pellets of trypanosomes were fixed briefly in glutaraldehyde and processed for thin section electron microscopy or freeze-cleave replicas. Observations of sectioned material facilitated orientation and comparison of details seen in replicas. Fracture faces of cell body and flagellar membranes as well as 3-dimensional views of the nuclear membrane were studied. Cell body membranes of 80% of the organisms from intact rats contained random arrays of intramembranous particles (IMP). Aggregated clusters of particles appeared on the fracture faces of 20% of the trypanosomes. Some of these membranes had nonrandomly distributed particles aligned in distinct rows on the outer fracture face of both cell body and flagellum. Many inner face fractures of the cell body membranes had a particle arrangement similar to the longitudinal alignment of cytoskeletal microtubules. No aggregated particle distribution was seen in membranes of trypanosomes harvested from lethally irradiated rats. Replicas of trypanosome pellets also had plasmanemes as a series of attached, empty, coated membrane vesicles. These structures were found in close association with, as well as widely separated from the parasites. The shedding of these vesicles and the variation of particles in cell body membranes are discussed in light of antibody-induced architectural and antigenic changes in surface properties of trypanosomatids. The convex face of the inner membrane of the nucleus also is covered with randomly arrayed particles. More IMP were observed on the inner than on the outer nuclear membranes. Images of nuclear pores were also seen. The importance of these structures in drug and developmental studies of trypanosomes is discussed. On fracture faces of the flagellar membrane there were miniature maculae adherentes, unique to the inner fracture face and occurring only at regions of membrane apposition between cell body and flagellum. Each cluster of particles exposed by the freeze-cleave method corresponds to an electron-dense plaque seen in thin section images. However, because of a unique fracture pattern, these plaques were not revealed on the apposing body membranes, as illustrated in thin sectioned organisms.     

Organic Carbon Utilization by Obligately and Facultatively Autotrophic Beggiatoa Strains in Homogeneous and Gradient Cultures

Marine Beggiatoa strains MS-81-6 and MS-81-1c are filamentous gliding bacteria that use hydrogen sulfide and thiosulfate as electron donors for chemolithotrophic energy generation. They are known to be capable of chemolithoautotrophic growth in sulfide gradient media; here we report the first successful bulk cultivation of these strains in a defined liquid medium. To investigate their nutritional versatilities, strains MS-81-6 and MS-81-1c were grown in sulfide-oxygen gradient media supplemented with single organic compounds. Respiration rates and biomass production relative to those of controls grown in unsupplemented sulfide-limited media were monitored to determine whether organic compounds were utilized as sources of energy and/or cell carbon. With cells grown in sulfide gradient and liquid media, we showed that strain MS-81-6 strongly regulates two enzymes, the tricarboxylic acid cycle enzyme 2-oxoglutarate dehydrogenase and the Calvin cycle enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase, in response to the presence of organic carbon (acetate) in the growth medium. In contrast, strain MS-81-1c lacked 2-oxoglutarate dehydrogenase activity and regulated ribulose-1,5-bisphosphate carboxylase/oxygenase activity only slightly in response to organic substrates. Tracer experiments with radiolabeled acetate showed that strain MS-81-1c did not oxidize acetate to CO(inf2) but could synthesize approximately 20% of its cell carbon from acetate. On the basis of these results, we conclude that Beggiatoa strain MS-81-1c is an obligate chemolithoautotroph, while strain MS-81-6 is a versatile facultative chemolithoautotroph.     

AN ELECTRON MICROSCOPE STUDY OF THE EARLY ASSOCIATION BETWEEN TWO MAMMALIAN VIRUSES AND THEIR HOSTS

Early interaction between two animal viruses, vaccinia and adenovirus 7, which multiply readily in L strain and HeLa cells, respectively, was examined in both whole mount preparations and in thin sections. To observe the association at the surface, cells carrying adsorbed virus were swelled under controlled conditions and then "stained" with neutral phosphotungstate. Each particle of both virus types becomes attached to the cell by several capsomeres and is then ingested by phagocytosis. Within the cell, near the surface, single particles or small clumps of adenovirus are lodged within vesicles. Deeper in the cytoplasm this virus is packed in large, numerous inclusions, whereas very close to the nuclear envelope only free particles are found. Vaccinia, on the other hand, either free or in vesicles, is always found in the cytoplasm, at some distance from the nucleus (11). Adsorption and intracellular disposition of these two viruses is discussed in relation to the infectious process.     

Macromolecular subunits on the surfaces of the cell wall of Thiocapsa roseopersicina]

A layer of fungiform macromolecular subunits was found on the surface of the cell wall of Thiocapsa roseopersicina, a purple sulphur bacterium, strain BBS. The cap of a particle has a diameter of 40 to 60 A; the stalk is 80 to 100 A long and 20 to 30 A thick. Under the conditions of nitrogen fixation and a low content of vitamin B12 (0.1 mcg/litre) in the cultural broth, a second layer of similar particles is formed over the first layer.     

The Fine Structure of Secretion in Hyalophysa chattoni: Formation of the Attachment Peduncle and the Chitinous Phoretic Cyst Wall

ABSTRACT. The settling tomite stage of the apostome Hyalophysa chattoni secretes a phoretic cyst wall composed of chitin, mucopolysaccharides, and protein. Within 1 1/2 h after settling, an electron-dense proteinaceous cyst layer (the outer layer) is formed from secretions originating at the base of the kineties and from the thick pellicular layer between the kineties. The inner cyst layer, composed primarily of chitin (acidic and neutral polysaccharides are also present), is secreted across the entire cell surface. Cyst wall formation is completed within 6 h. The fine structure of endocyst secretion resembles stages in the secretion of chitin by fungi, yeasts, and arthropods. A proteinaceous attachment peduncle is secreted to anchor the cell to a shrimp host and is formed by the release of electron-dense secretory bodies from the cell's ventral surface.