MORPHOLOGICAL CHANGES OF BLOOD PLATELETS IN MAN DURING DEEP SATURATION DIVING

1987 ◽  
Author(s):  
Hanne Klausen ◽  
Per Flood ◽  
John O Hjelle ◽  
Holm Holmsen
Keyword(s):  
Morphological Changes ◽  
Blood Platelets ◽  
Venous Blood ◽  
Gas Bubbles ◽  
En Bloc ◽  
Deep Diving ◽  
Transmission Electron ◽  
Cacodylate Buffer ◽  
High Incidence

The small number of reports concerning blood platelets during deep saturation diving have shown that there may be a decrease in the number of circulating platelets. Gas bubbles are often present in theblood during decompression. In vitro, gas bubbles activate platelets, and alter the shape from discoidto spherical configuration with multiple, blunt spikes. This study was done to investigate if there are changes in the morphology of human blood platelets during deep diving. An 18 day long experimentalon-shore saturation dive to 360 msw was performed at NUTEC 1986. Bloodsamples were obtained from the 6 divers on 6 occations: pre-dive control, at 360 msw, 300 msw, 140 msw, 1 and 3 days after surfacing. Using 18 G Wasserman needles antecubital venous blood samples (3ml) were collected directly into the fixative agent (7 ml of 2% glutardialdehyde in cacodylate buffer) whilestill in the hyperbaric chamber. The samples were then decompressed,and the platelets separated from the blood by centrifugation at roomtemperature for 10 min at 190 g. Preparation to transmission electron microscopy included post-fixation in osmium tetroxide, staining uranyl-en-bloc and eventually lead, dehydration and embedding in Epon.Ultrathin sections were examined by a Philips 300 I electron microscope. The examination revealed alterations in both platelet size and shape in the course of the dive. The mean platelet area was increased during and immidiately after thedive, the greatest increase occuring at 360 msw. There was a high incidence of shape changed plateletswith spherical configuration and multiple, blunt spikes. This form was by far most abundant at 360 msw, and the morphology normalised towards surface. This rises the suspection of a pressure-related rather rhan bubble-related effect and the results indicate that plateletsin circulation can be activated bypressure itself

Ultrastructure of E. coli: An in vitro study of cells cultured in the presence of four gastrointestinal (GI) hormones

1990 ◽  
Vol 48 (3) ◽  
pp. 628-629
Author(s):  
Leo J. Henz ◽  
Frank E. Johnson
Keyword(s):  
Morphological Changes ◽  
In Vitro Study ◽  
Uranyl Acetate ◽  
Culture Collection ◽  
En Bloc ◽  
Thin Sections ◽  
E Coli ◽  
Cacodylate Buffer ◽  
Synthetic Hormones

Hormones are found in exocrine secretions entering the gut. They alter the morphology of many eukaryotic cells; whether they affect the morphology of enteric flora is unknown. In this study, we examined the ultrastructure of E. coli, a common bacterium in the mammalian gut, for morphological changes resulting from exposure to GI hormones.E. coli (#11775 from American Type Culture Collection) were grown in protease-free trypticase soy broth (TSB) at 37°C for 18 hr to a concentration of 2 x 107 cells/ml. Pure synthetic hormones were used: sulfated C-terminal cholecystokinin octapeptide (CCK), pentagastrin (PG), cyclic somatostatin tetradecapeptide (SS), or the porcine form of secretin (SEC). These were individually added to. bacterial cultures in TSB to make 1 x 107 organisms/ml and 0.0, 0.5, 2.5, or 5.0 μg of hormone/ml, then incubated for 30 min at 37°C. The cultures were rapidly chilled and added to equal volumes of cold 6% glutaraldehyde in 0.2 M cacodylate buffer. After 30 min, the bacteria were concentrated by centrifugation (15 min at 4000 RPM) and the pellets suspended in cold 3% glutaraldehyde for an additional 15 min, followed by centrifugation. The pellets were resuspended in cold cacodylate buffer and stored at 2°C for 1-7 d. The cells were again centrifuged and the pellets were blotted with a strip of filter paper to remove excess fluid, then mixed with a drop of warm 2% agar. The agar suspensions were pipetted into cold saline. The resulting solidified extrusions were cut by hand into 2 mm segments for further processing in 1% OsO4 (with or without en bloc staining in 2% uranyl acetate (UA) in ethanol). Following dehydration in ethanol, rinsing in propylene oxide, and encapsulation in Epon-Araldite, thin sections were examined and photographed with a JEOL-100C microscope.

Correlation Between Cellular Functions and Morphological Changes of Human Trophoblast in Vitro

1975 ◽  
Vol 33 ◽  
pp. 600-601
Author(s):  
John C. Garancis ◽  
Robert O. Hussa ◽  
Michael T. Story ◽  
Donald Yorde ◽  
Roland A. Pattillo
Keyword(s):  
Electron Microscopy ◽  
Cellular Proliferation ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Cellular Functions ◽  
Human Trophoblast ◽  
Transmission Electron ◽  
Marked Activity ◽  
Malignant Trophoblast

Human malignant trophoblast cells in continuous culture were incubated for 3 days in medium containing 1 mM N6-O2'-dibutyryl cyclic adenosine 3':5'-monophosphate (dibutyryl cyclic AMP) and 1 mM theophylline. The culture fluid was replenished daily. Stimulated cultures secreted many times more chorionic gonadotropin and estrogens than did control cultures in the absence of increased cellular proliferation. Scanning electron microscopy revealed remarkable surface changes of stimulated cells. Control cells (not stimulated) were smooth or provided with varying numbers of microvilli (Fig. 1). The latter, usually, were short and thin. The surface features of stimulated cells were considerably different. There was marked increase of microvilli which appeared elongated and thick. Many cells were covered with confluent polypoid projections (Fig. 2). Transmission electron microscopy demonstrated marked activity of cytoplasmic organelles. Mitochondria were increased in number and size; some giant forms with numerous cristae were observed.

Paracrystalline bundles of large tubules, induced in vitro by Mebendazole in Cysticercus cellulosae

Parasitology ◽  
1981 ◽  
Vol 83 (3) ◽  
pp. 513-518 ◽  
Author(s):  
J. P. Laclette ◽  
Marie Therese Merchant ◽  
Kaethe Willms ◽  
L. Cañedo
Keyword(s):  
Electron Microscopy ◽  
Culture Medium ◽  
Morphological Changes ◽  
Bladder Wall ◽  
Secretory Cells ◽  
External Diameter ◽  
Nematode Larvae ◽  
Transmission Electron ◽  
Cysticercus Cellulosae

SUMMARYThe effect of the anthelmintic Mebendazole on Cysticercus cellulosae maintained in culture medium was studied by transmission electron microscopy. In addition to the well-known morphological changes induced by Mebendazole in other cestode and nematode larvae, it also induced the cytoplasmic appearance of paracrystalline bundles in the secretory cells of the bladder wall. These bundles were formed by groups of large parallel tubules arranged in a hexagonal-like pattern. The tubules, which had an external diameter of about 50 nm and a length that might exceed 5 μm, were surrounded by a matrix and a distance between neighbouring tubules of 80–120 nm centre to centre was estimated. The tubules were stable to colchicine and low temperature. The temporary appearance of bundles is described and some alternative explanations on their origin are advanced.

scholarly journals Differentiation of a teratocarcinoma line: preferential development of cholinergic neurons.

1981 ◽  
Vol 88 (1) ◽  
pp. 57-66 ◽  
Author(s):  
S E Pfeiffer ◽  
H Jakob ◽  
K Mikoshiba ◽  
P Dubois ◽  
J L Guenet ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Cholinergic Neurons ◽  
Carcinoma Cells ◽  
Sodium Influx ◽  
Average Cell ◽  
Transmission Electron ◽  
Neuronal Processes ◽  
Embryonic Antigen ◽  
Average Cell Volume

A line of embryonal carcinoma cells, PCC7-S, established in vitro from a spontaneous testicular teratocarcinoma, has been studied. Upon removing the cells from a low density monolayer culture system and permitting the cells to form aggregates in suspension, we observed a change of several physical and biochemical parameters: (a) reduction in average cell volume, (b) blockage and accumulation of cells in G1, (c) rise in secreted protease activity, (d) rise in acetylcholinesterase and choline acetyltransferase activities, and (e) disappearance of embryonic antigen F9. Although PCC7 aggregates did not undergo substantial morphological changes while suspended, when aggregates 4 or more days old were allowed to attach to plastic tissue culture dishes, substantial neurite outgrowth occurred over the next 1-3 d. This process was markedly enhanced by the addition to the growth medium of carboxymethylcellulose and inhibitors of DNA synthesis. Transmission electron microscopy disclosed a neurite ultrastructure consistent with that of neuronal processes. A veratridine-stimulated, tetrodotoxin-blocked sodium influx of 100 nmol/min per mg protein was also observed in these differentiated surface cultures. This cell line is discussed in terms of its utility for the study of early events leading to a commitment to cellular differentiation, as well as for the investigation of terminal differentiation to cholinergic neurons.

Ultrastructural Characterization of Platelet-Activating Factor-Stimulated Human Eosinophils from Patients with Asthma

1993 ◽  
Vol 84 (4) ◽  
pp. 391-399 ◽  
Author(s):  
Claus Kroegel ◽  
Ann Dewar ◽  
Tatsuo Yukawa ◽  
Per Venge ◽  
Peter J. Barnes ◽  
...  
Keyword(s):  
Morphometric Analysis ◽  
Morphological Changes ◽  
Shape Change ◽  
Platelet Activating Factor ◽  
Eosinophil Cationic Protein ◽  
Cell Periphery ◽  
Cationic Protein ◽  
Asthmatic Patients ◽  
Transmission Electron

1. Purified human eosinophils from asthmatic patients were stimulated with platelet-activating factor in vitro and examined for morphological changes by transmission electron and light microscopy. Changes were also evaluated by morphometric analysis and were related to the platelet-activating factor-stimulated release of granular eosinophil cationic protein. 2. Stimulation of eosinophils with platelet-activating factor induced a dose-dependent shape change, including the elongation of cells, loss of microvilli and the formation of lamellipodia. This effect was maximal at 25 min and was reversible. 3. Stimulation with platelet-activating factor also induced granule movement to the cell periphery and fusion of adjacent granules. Granules became swollen and vesiculated, whereas both the matrix and core showed evidence of solubilization. 4. There was a time-dependent secretion of eosinophilic cationic protein from human eosinophils upon stimulation with platelet-activating factor which occurred without significant lactate dehydrogenase release. 5. Morphometric analysis of the transmission electron micrographs indicated a significant reduction in cytoplasmic area after 10 min of incubation with platelet-activating factor from 39.0 ± 1.7 μm2 for untreated eosinophils to 33.2 ± 2.3 μm2 (P < 0.02) for platelet-activating factor-treated cells, underscoring the observation that the cells change from spherical to ellipsoidal. No significant increase in the perimeter of the cells was found. 6. The number of granule-profiles in platelet-activating factor-stimulated eosinophils was slightly reduced when compared with control, and an increase in granule area was observed 10 min after platelet-activating factor challenge (0.215 ± 0.011 μm2 versus 0.246 ± 0.016 μm2). 7. Human eosinophils from patients with asthma stimulated with platelet-activating factor undergo both cellular and granular alterations and reorganization which parallel the release of granular eosinophil basic protein.

scholarly journals Antileishmanial Activity of a Linalool-Rich Essential Oil from Croton cajucara

2003 ◽  
Vol 47 (6) ◽  
pp. 1895-1901 ◽  
Author(s):  
Maria do Socorro S. Rosa ◽  
Ricardo R. Mendonça-Filho ◽  
Humberto R. Bizzo ◽  
Igor de Almeida Rodrigues ◽  
Rosangela Maria A. Soares ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Essential Oil ◽  
Mammalian Cells ◽  
Morphological Changes ◽  
Peritoneal Macrophages ◽  
Transmission Electron ◽  
Low Concentrations ◽  
Mouse Peritoneal Macrophages ◽  
Croton Cajucara

ABSTRACT The in vitro leishmanicidal effects of a linalool-rich essential oil from the leaves of Croton cajucara against Leishmania amazonensis were investigated. Morphological changes in L. amazonensis promastigotes treated with 15 ng of essential oil per ml were observed by transmission electron microscopy; leishmanial nuclear and kinetoplast chromatin destruction, followed by cell lysis, was observed within 1 h. Pretreatment of mouse peritoneal macrophages with 15 ng of essential oil per ml reduced by 50% the interaction between these macrophages and L. amazonensis, with a concomitant increase by 220% in the level of nitric oxide production by the infected macrophages. Treatment of preinfected macrophages with 15 ng of essential oil per ml reduced by 50% the interaction between these cells and the parasites, which led to a 60% increase in the amount of nitric oxide produced by the preinfected macrophages. These results provide new perspectives on the development of drugs with activities against Leishmania, as linalool-rich essential oil is a strikingly potent leishmanicidal plant extract (50% lethal doses, 8.3 ng/ml for promastigotes and 8.7 ng/ml for amastigotes) which inhibited the growth of L. amazonensis promastigotes at very low concentrations (MIC, 85.0 pg/ml) and which presented no cytotoxic effects against mammalian cells.

scholarly journals Theβ-SiC Nanowires (~100 nm) Induce Apoptosis via Oxidative Stress in Mouse Osteoblastic Cell Line MC3T3-E1

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Weili Xie ◽  
Qi Xie ◽  
Meishan Jin ◽  
Xiaoxiao Huang ◽  
Xiaodong Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Oxidative Dna Damage ◽  
Morphological Changes ◽  
Osteoblastic Cell ◽  
Osteoblastic Cell Line ◽  
Sic Nanowires ◽  
Tissue Replacement ◽  
Transmission Electron ◽  
First Time

Silicon carbide (SiC), a compound of silicon and carbon, with chemical formula SiC, the beta modification (β-SiC), with a zinc blende crystal structure (similar to diamond), is formed at temperature below1700∘C.β-SiC will be the most suitable ceramic material for the future hard tissue replacement, such as bone and tooth. Thein vitrocytotoxicity ofβ-SiC nanowires was investigated for the first time. Our results indicated that 100 nm long SiC nanowires could significantly induce the apoptosis in MC3T3-E1 cells, compared with 100 μm long SiC nanowires. And 100 nm long SiC nanowires increased oxidative stress in MC3T3-E1 cells, as determined by the concentrations of MDA (as a marker of lipid peroxidation) and 8-OHdG (indicator of oxidative DNA damage). Moreover, transmission electron microscopy (TEM) was performed to evaluate the morphological changes of MC3T3-E1 cells. After treatment with 100 nm long SiC nanowires, the mitochondria were swelled and disintegrated, and the production of ATP and the total oxygen uptake were also decreased significantly. Therefore,β-SiC nanowires may have limitations as medical material.

Henna: A Potential Cause of Oxidative Hemolysis and Neonatal Hyperbilirubinemia

PEDIATRICS ◽  
1996 ◽  
Vol 97 (5) ◽  
pp. 707-709
Author(s):  
William H. Zinkham ◽  
Frank A. Oski
Keyword(s):  
Reduced Glutathione ◽  
Venous Blood ◽  
Oxidative Potential ◽  
Oxidative Hemolysis ◽  
High Incidence ◽  
The World ◽  
Before And After ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Cutaneous Application

Objective. To evaluate the in vitro oxidative potential of lawsone (2-hydroxy-1,4 naphthoquinone). Lawsone is a chemical present in henna, the crushed leaves of which are used worldwide as a cosmetic agent to stain the hair, skin, and nails. Methodology. Venous blood from glucose-6-phosphate dehydrogenase (G6PD)-normal and G6PD A- subjects were incubated with various amounts of lawsone for 2 hours at 37°C. Reduced glutathione and methemoglobin (MHb) levels were measured before and after incubation. Results. Final molar concentrations of lawsone in normal blood of 1.4, 2.8, 5.7, and 8.6 x 10-3 mol/L increased MHb percentages from 0.5% to 2.2%, 8.3%, 9.5%, and 12.5%, respectively. In a G6PD A- blood, MHb percentages were 19.8%, 32.2%, 44.9%, and 53.9%. At a lawsone concentration of 2.8 x 10-3 mol/L, blood from 15 healthy adults formed MHb percentages of 7.4% ± 3.3% (±1 SD); in blood from 4 G6PD A- adults, percentages were 44.5%, 40.6%, 41.3%, and 42.8%. Simultaneous measurements of reduced glutathione revealed preincubation values of greater than 40 mg/100 mL of red cells in blood of healthy and G6PD A- subjects. Postincubation values were greater than 40 in blood of healthy subjects and less than 40 in blood of G6PD A- subjects. Conclusions. These in vitro observations indicate that lawsone is an agent capable of causing oxidative hemolysis. In regions of the world where there is a high incidence of G6PD deficiency and unexplained hyperbilirubinemia, oxidative hemolysis secondary to the cutaneous application of henna could be the initiating event.

Effect of serum on the mitochondrial active area on developmental days 1 to 4 in in vitro-produced bovine embryos

Zygote ◽  
2011 ◽  
Vol 19 (4) ◽  
pp. 297-306 ◽  
Author(s):  
M. Crocco ◽  
R.H. Alberio ◽  
L. Lauria ◽  
M.I. Mariano
Keyword(s):  
Electron Microscopy ◽  
Culture Medium ◽  
Morphological Changes ◽  
Mitochondrial Dynamics ◽  
Developmental Changes ◽  
Bovine Embryos ◽  
Functional Changes ◽  
Transmission Electron ◽  
Subcellular Level

SummaryCertain morphological changes at the subcellular level caused by the current techniques for in vitro embryo production seem to affect mitochondria. Many of these, including dysfunctional changes, have been associated with the presence of serum in the culture medium. Thus, the aim of the present work was to assess the mitochondrial dynamics occurring in embryos during the first 4 days of development, in order to analyze the most appropriate time for adding the serum. We used transmission electron microscopy (TEM) micrographs to calculate the embryo area occupied by the different morphological types of mitochondria, and analyzed them with Image Pro Plus analyzer. The results showed hooded mitochondria as the most representative type in 1- to 4-day-old embryos. Swollen, on-fusion, orthodox and vacuolated types were also present. When analyzed in embryos cultured without serum, the dynamics of the different mitochondrial types appeared to be similar, a fact that may provide evidence that the developmental changes control the mitochondrial dynamics, and that swollen mitochondria may not be completely inactive. In contrast, in culture medium supplemented with serum from estrous cows, we observed an increased area of hooded mitochondria by developmental day 4, a fact that may indicate an increased production of energy compared with previous days. According to these results, the bovine serum added to the culture medium seems not to be responsible for the functional changes in mitochondria.

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