An apparatus for cytophotometry; determination of the relative degree of basophilia of cell structures with special reference to the nerve cells.
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The article is based on newly published data concerning cell structure (cytoskeleton and cytoplasmic organelle) and biochemistry of the protozoon Toxoplasma gondii, many of which can be applied in therapy of toxoplasmosis.
The thermal death mechanism of microorganisms when heated at lethally high temperatures is still not fully understood. In this study, we examined the relationship between thermal death and degradation of the cell structure in the mesophilic bacterium Escherichia coli strain W3110 and the hyperthermophilic archaeon Thermococcus sp. strain Tc-1-95. By heating the microorganisms at lethally high temperatures only briefly (1.5 s duration) in a flow-type apparatus, we studied the microbial cells at very early and critical stages of the thermal death process. For E. coli, it was found that the loss of viability was not associated with thermal damage to the cell envelope. Deformation of the nucleoid was observed. These results suggest that the thermal death of E. coli is attributed to thermal denaturation or degradation of cytoplasmic molecules. On the other hand, the thermal death of Thermococcus sp. strain Tc-1-95 was strongly associated with rupture of the cell envelope. Furthermore, massive deformation of the S-layer with lethal thermal stress was observed. These results demonstrate that the thermal deaths of the two microorganisms investigated proceed via very different mechanisms. The contrast can be attributed to the difference in their cell envelope structures.
It was found that a slide centrifuge (Cytospin) preparation of a cell suspension allowed a reliable assessment of not only cell structure but also the percentage of non-viable cells. The non-viable cells appeared as "smear" cells and paralleled in number the cells taking up trypan blue. Direct experiment showed the unstained viable cells in a trypan blue cell suspension remained intact in a Cytospin preparation while the cells taking up trypan blue were the "smear" cells. The non-viability of the "smear" cells was confirmed by their inability to survive in culture.
Structural changes which occur in the Malpighian tubule yellow region primary cells during larval-pupal-adult development of the skipper butterfly Calpodes ethlius are described. The developmental changes in cell structure are correlated with functional changes in fluid transport (Ryerse, 1978a) in a way which supports osmotic gradient models of fluid secretion. Larval tubules are specialized for fluid secretion with deep basal infolds and elongate mitochondria-containing apical microvilli which provide channels in which osmotic gradients could be set up. The Malpighian tubule cells are extensively remodelled at pupation when fluid transport is switched off, but they persist intact through metamorphosis. At this time, the basement membrane doubles in thickness, the mitochondria are retracted from the microvilli and are isolated for degradation in autophagic vacuoles, and both apical and basal plasma membranes are internalized via coated vesicles for degradation in multivesicular bodies, which results in the shortening of the microville and the disappearance of the basal infolds. Mitochondria are re-inserted into the microvilli, and the basal infolds re-form in pharate adult stage Malpighian tubules when fluid secretion resumes. Adult tubules are similar in general structure to larval tubules and contain mitochondria in the microvilli and basal infolds. However, they differ from larval tubules in that they are capable of very rapid fluid transport, have a reduced tubule diameter and tubule wall thickness, a much thicker basement membrane and peripherally associated tracheoles. Mineral concretions of calcium phosphate accumulate in larval tubules, persist through metamorphosis and decline in number in adults, suggesting they serve some anabolic role.
Studying exogenously expressed tagged proteins in live cells has become a standard technique for evaluating protein distribution and function. Typically, expression levels of experimentally introduced proteins are not regulated, and high levels are often preferred to facilitate detection. However, overexpression of many proteins leads to mislocalization and pathologies. Therefore, for normative studies, moderate levels of expression may be more suitable. To understand better the dynamics of intermediate filament formation, transport, and stability in a healthy, living cell, we inserted neurofilament heavy chain (NFH)-green fluorescent protein (GFP) fusion constructs in adenoviral vectors with tetracycline (tet)-regulated promoters. This system allows for turning on or off the synthesis of NFH-GFP at a selected time, for a defined period, in a dose-dependent manner. We used this inducible system for live cell imaging of changes in filament structure and cell shape, motility, and transport associated with increasing NFH-GFP expression. Cells with low to intermediate levels of NFH-GFP were structurally and functionally similar to neighboring, nonexpressing cells. In contrast, overexpression led to pathological alterations in both filament organization and cell function.
Flavobacterium psychrophilum, the pathogen of bacterial cold-water disease, causes serious problems in ayu Plecoglossus altivelis culture. This study investigated the effect of the culture period of F. psychrophilum and on the structure of its cells. From the SDS-PAGE of total proteins of cellular components, much difference was found between the 36 hr culture and the 48 and 72 hr cultures. A SEM observation of the cells showed many fragments, especially on the cell surface of the 36 hr culture. These fragments consisted of an outer membrane, seen by TEM observation, and may contain substances causing the virulence. Specific proteins observed by the SDS-PAGE and fragments in the 36 hr culture may be related to the virulence of F. psychrophilum.
OBJECTIVE: To study the cell structure changes of Gastrodia elata after Armillaria mellea infection as well as the nutritional resource of this important medicinal plant. METHOD: Observation of the serial sections on the nutritional stems of G. elata, and measure new corms of G. elata when cut off the old corms connection with rhizomorph of A. mellea. RESULT: After G. elata nutritional stems infected by A. mellea, the rhizomorph of A. mellea separate into several hyphal layers and penetrate hyphal stream which infect into the cells of cortex layer in the direction of outside and infect directly into layer cells in the direction of inside, these hypha can be used the nutrition of G. elata the new corms will be stop growth if cut off the rhizomorph of A. mellea connected with G. elata. CONCLUSION: After the rhizomorph of A. mellea infected G. elata nutritional stems, hyphal coils and the hyphal stream will breakthrough passage cells as well as large cells surround the whole nutritional stems, so the passage cells is the key nutritional resource in the whole growth period of G. elata.
Advances in two techniques have made the problem of assessing the acute and/or chronic effects of toxic agents on Leydig cell structure and testosterone synthesis and secretion amenable to study. First, in vitro testicular perfusion has been perfected to a point where it closely resembles in situ testosterone secretion. Second, now it is possible to quantify the proportion of Leydig cell cytoplasm occupied by the cellular organelles which contain steroidogenic enzymes. Herein, we report that inhibition of Leydig cell steroidogenic enzymes is reflected by reduced testosterone secretion by in vitro perfused rat and rabbit testes. Moreover, the activity of specific steroidogenic reactions can be monitored by measuring the secretion of reaction substrate(s) and product(s) from in vitro perfused testes. Testosterone secretion by in vitro perfused testes from five species is highly and positively correlated with the volume density of smooth endoplasmic reticulum in Leydig cell cytoplasm. Exploitation of these findings will allow toxicologists to quantitatively assess the effect of toxicants on Leydig cell testosterone biosynthesis and secretion, to identify the specific steroidogenic enzymes affected, to assess whether the membranous environment of the steroidogenic enzymes is compromised, and perhaps even to predict the deleterious effect of a toxic agent on Leydig cell steroidogenic function from a stereological assessment of Leydig cell ultrastructure.
Haemagglutinating activity was determined in cell walls and total cell organelles of crown cells of Winter wheat (Triticum aestivum L. ) plants. The effect of fusicoccin (FC) was investigated using fractions obtained from plants hardened for 7 days at 2 degrees C and from untreated plants. FC concentration (5x10(-7) m) increased the frost resistance of the plants. The temporal pattern of lectin activity during hardening could be described by a single-peak curve. In the cell wall fraction, the highest activity manifested itself after one-day hardening, and in the fraction of organelles it peaked after five-days hardening. The carbohydrate specificity of lectins also changed during hardening; cell wall lectins completely lost their capacity for interaction with uridine diphosphoglucose, glucose 6-phosphate, D-galactosamine, and N-acetylglucosamine and the lectins of organelles retained some affinity only for amino sugars. After hardening the test plants, the activity of the lectins increased substantially in the cell walls and plastids, decreased in the nuclei, and was practically flat in mitochondria and microsomes. Consequently, low temperature and FC with their antistress effect improved frost resistance and stimulated the activity of the lectins of some cell structures of the tillering node of winter wheat. A similar action of low temperature and FC in increasing the activity of lectins of plastids was found. Further information was obtained on the subcellular localization of lectins providing additional information on their possible participation in the development of frost resistance of winter wheat.
To determine the effect of intracoronary perfluorochemical on endothelial cell structure and function, 16 dogs were randomized to receive either low-dose (15 ml/kg) intracoronary perfluorochemical (Fluosol-DA) or saline after 90 min of proximal occlusion of the left anterior descending coronary artery (LAD). The animals underwent reperfusion for 60 min with the introduction of perfluorochemical or saline 5 to 10 min after the onset of reperfusion. Endothelium-dependent coronary vasodilatory reserve was determined in vivo both at baseline and 1 hr after reperfusion by infusion of acetylcholine and then serotonin into the distal LAD bed in 12 animals (six in each group). Both agonists significantly increased regional flow measured by 133Xe washout in the two groups before occlusion, but at 1 hr after reperfusion only animals given perfluorochemical demonstrated a significant increase in flow. Vasodilatory reserve was assessed in vitro with cumulative dose-response curves to acetylcholine on LAD rings proximal and distal to the snare in all animals. These studies demonstrated a significant reduction in endothelial cell-mediated relaxation of epicardial arterial segments in the ischemic segment of control but not treated animals. Light microscopy revealed the presence of neutrophils within vessels in the ischemic zones in control animals only. Electron microscopy showed capillary obstruction by endothelial cell protrusions and neutrophil and red cell plugging in control animals in the ischemic region but an intact endothelium and predominantly unobstructed capillaries in treated animals. These findings suggest that the structural and functional endothelial damage after reperfusion may be prevented by the administration of intracoronary perfluorochemical after the onset of reperfusion.
Incorporation of 3H-leucine into proteins of rat brain cell structures during application of antifeins (compounds of alternative action on memory processes) has been studied. No correlation was observed between changes in protein synthesis in nuclei, mitochondria, components of endoplasmic reticulum and memory effects of ethyl-, allyl- and propylnorantifeins. Only M1 and M2-demethylated structural analogs of ethylnorantifeins (exerting the most effective action on RNA synthesis and retention of conditional reflexes) enhanced the synaptosomal protein synthesis.
The study presented in this article investigated the influence of different Cr(III) and Cr(VI) compounds in the cultivation medium on the uptake and localization of chromium in the cell structure of the yeast Candida intermedia. The morphology of the yeast cell surface was observed by the scanning electron microscopy. Results demonstrated that the growth inhibitory concentration of Cr(III) in the cultivation medium induced changes in the yeast cell shape and affected the budding pattern, while inhibitory concentration of Cr(VI) did not cause any visible effects on morphological properties of the yeast cells. The amount of total accumulated chromium in yeast cells and the distribution of chromium between the yeast cell walls and spheroplasts were determined by atomic absorption spectroscopy. No significant differences were found neither in total chromium accumulation nor in the distribution of chromium in yeast cell walls and spheroplasts between the two of Cr(VI) compounds. Conversely, substantial differences between Cr(III) compounds were demonstrated in the total uptake as well as the localization of chromium in yeast cells.
The cytokine IL-7 and its receptor are essential for normal B and T lymphopoiesis. We have analyzed the role of this receptor in B cell development throughout ontogeny in IL-7 receptor alpha-deficient mice. We demonstrate that the IL-7 receptor becomes progressively more important with age. B lymphopoiesis takes place, albeit at reduced levels, in fetal liver and bone marrow of young mice, but is arrested in adults. The outcome is a severe reduction, from an early age, in peripheral B cells including follicular, marginal zone and B-1 B cells as well as perturbed splenic B cell structures, which are restored after adoptive transfer of normal spleen cells. We conclude that in the absence of the IL-7 receptor, the residual B lymphopoiesis occurring early in ontogeny must be facilitated by another component, whereas the IL-7 receptor is the key factor in adults. The impairment of marginal zone and B-1 B cells in IL-7 receptor- but not IL-7-deficient mice suggests non-redundant functions for the IL-7 receptor ligands, IL-7 and thymic stromal lymphopoietin.
The effects of tebuconazole, a systemic fungicide, on the morphology, structure, cell wall components and toxin production of Fusarium culmorum were investigated in vitro. Treatment was by application of four filter paper strips (0.75 cm x 5.0 cm) soaked in 20 micrograms ml-1 fungicide placed around a point inoculum in Petri dishes. Mycelial growth was strongly inhibited by fungicide treatment. Scanning electron microscopic observations showed that the fungicide caused irregular swelling and excessive branching of hyphae. The morphological changes induced by the fungicide at the ultrastructural level included considerable thickening of the hyphal cell walls, excessive septation, the formation of the incomplete septa, extensive vacuolisation, accumulation of lipid bodies and progressing necrosis or degeneration of the hyphal cytoplasm. Non-membrane inclusion bodies were often detected in the hyphal cytoplasm. Furthermore, the formation of new hyphae (daughter hyphae) inside collapsed hyphal cells was common following treatment. The daughter hyphae also displayed severe alterations such as irregular thickening of the cell walls and necrosis of the cytoplasm. Using cytochemical techniques, the labelling densities of chitin and beta-1,3-glucan in the cell walls of the fungicide-treated hyphae were more pronounced than in those of the control hyphae. Moreover, immunogold labelling with antiserum against deoxynivalenol (DON) revealed that Fusarium toxin DON was localized in the cell walls, cytoplasm, mitochondria and vacuoles of the hyphae from the control and the fungicide treatment, but the labelling density in the fungicide-treated hyphae decreased dramatically compared with the control hyphae, indicating that tebuconazole reduced Fusarium toxin production of the fungus.
This report discusses fundamental limitations in attempting to derive cell size, shape, or distribution from the two-dimensional images provided by conventional electron microscopy. Morphometric or stereologic measurement of random thin sections is a convenient way to obtain some information of this type. However, it cannot provide complete, objective information about real size, shape, or connectivity of cells containing irregular or unevenly distributed structures or nonuniform populations of cells. Anisotropic structures require analysis of a complete set of serial sections. The analysis may utilize either stereo, mono, or tilted optical slices, and subsequent integration of this information into a single 3-D computer data set. In this study, we analyze stereo pairs of high-voltage electron micrographs of serial thick sections (0.5 micron) and critical-point-dried whole-cell mounts of rat brain astroglial cell cultures. The Z-axis resolution is increased by digitizing contours at discrete levels within each stereo view. This is accomplished with a new type of stereoscopic contouring device. We calculated area and volume changes accompanying hypo-osmolar swelling and spontaneous reversal of the swelling. (Regulatory Volume Decrease-RVD). An understanding of the mechanism of swelling of astroglial cells is important for improving the treatment of brain injury. The total cell-volume results are comparable with results previously obtained using nonmetabolized, radioactively tagged compounds that diffuse into various cell compartments. Our serial-section and whole-cell data also provide new information about the relative swelling of nucleus, cytoplasm, and individual organelles such as mitochondria. The basic biological problem being approached is whether homeostasis of cell function is accompanied by surface area and volume regulation of enzyme-rich membranes and organelles. Conversely, it is proposed to explore the possibility that abnormal organelle areas and volumes are indicators of perturbations of cell division, metabolism, or gene expression.
Schaefer, Werner B. (National Jewish Hospital, Denver, Colo.), and C. Willard Lewis, Jr. Effect of oleic acid on growth and cell structure of mycobacteria. J. Bacteriol. 90:1438-1447. 1965.-The growth-stimulatory effect of fatty acids on Mycobacterium kansasii and other mycobacterial species is associated with transient changes in the morphology of the bacteria. One change observed is the appearance of intracellular vacuoles separated by chromatinic crossbars. Evidence is presented that these changes are due to the rapid metabolic uptake and the accumulation of lipid in the form of globules. This process provides the cells with an internal reservoir of a preferred carbon and energy source. In certain mycobacterial species, including M. kansasii, the morphological changes are elicited by free and by esterified fatty acid; in others, only by free fatty acid. The latter strains apparently lack the enzyme to split the fatty acid ester.
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