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Biomedical subjects

R D Allen

Publications and source records attributed to R D Allen.

At least 217 records · Page 12Linked to original sources

Axenic Paramecium caudatum. I. Mass culture and structure.

To establish and grow Paramecium caudatum in mass axenic culture the culture medium of Soldo, Godoy & van Wagtendonk was modified by substituting phosphatidylethanolamine (PE) for TEM-4T and by a 10-fold increase in folic acid. Population densities of 4000 to 6000 cells/ml and a generation time of 20-26 h are regularly obtained. Optimal growth is obtained with PE-stigmasterol ratios between 40:1 to 400:1. Cells from 1-day-old axenic cultures have many lipid bodies aggregated in clumps (which disappear in 2 to 3 days) as well as foci of rough endoplasmic reticulum bordered by dictyosomes. The latter suggests a very active metabolism. Crystalline sheets found in both food vacuoles and lysosomes presumably play a role in digestion. Axenically grown cells also have abundant Golgi bodies (dictyosomes) and by late log phase become filled with lysosomes.

Animals↗

Structure, protein composition and birefringence of the costa: a motile flagellar root fibre in the flagellate Trichomonas.

The costa is a rod-shaped intracellular organelle lying in the cytoplasm immediately below the undulating membrane. In certain large species of Trichomonas (T. gigantea, T. termopsidis and an undescribed species from the termite Porotermes adamsoni) this organelle is motile. Bending waves are transmitted along the length of the costa, in either direction. It is shown that the bending is actively produced by the costa itself. The form of the movements is described in detail. The costa is birefringent. At the point at which bending occurs there is a sharply localized reduction in birefringence. Electron microscopy shows that the costa is composed of longitudinally running lamellae, 2--3 nm thick and spaced 12 nm apart. These are connected to transversely running elements spaced at intervals of about 37 nm. The lamellae occur in two alternative configurations: straight and zig-zag. Bending probably results from a local, transient change from the straight to the zig-zag configuration. This would account for the local change in birefringence which accompanies bending. Polyacrylamide gel electrophoresis of isolated costas shows them to contain a number of protein components, of which the principal one has a molecular weight of about 90,000. Preliminary cytochemical evidence is presented for ATPase activity in the costa. The costa is a type of motile system distinct from any hitherto described.

Adenosine Triphosphatases↗

Membrane recycling at the cytoproct of Tetrahymena.

Exocytosis and membrane recycling at the cytoproct (cell anus) of Tetrahymena pyriformis were studied using thin-section electron microscopy. Single cells were fixed at specific times relative to the elimination of the vacuole's contents--before elimination, at elimination, 3--5 s and 10--15 s following elimination. The closed cytoproct is distinguished from other pellicular regions by a single membrane at the cell surface which is circumscribed by an electron-opaque flange that links or welds the plasma membrane to the underlying alveolar margins. Microtubules originating in the flange pass inward where they lie over, and possibly guide, the approaching food vacuoles to the cytoproct. Food facuoles near the cytoproct are also accompanied by coats of microfilaments. These microfilaments appear to be active in the channelling and endocytosis of food vacuole membrane. Upon cytoproct opening the plasma membrane and food vacuole membrane fuse. Elimination seems to be essentially passive and is accomplished by re-engulfment of the old food vacuole membrane which is constantly associated with microfilaments. Reengulfment of all the food vacuole membrane requires 10--15 s and results in a closed cytoproct. The membrane remnants embedded in microfilaments form a cluster under the closed cytoproct. At the periphery of this cluster remnants take the shape of 70--130-nm spherical vesicles or 0.2-micrometer-long flattened vesicles.

Animals↗

Hand-mirror cell leukemia associated with mental retardation: immunologic, chromosome, and morphological studies.

Cycochemical, morphological, immunologic, and cytogenetic studies were carried out on hand-mirror cells (HMC) from a mentally retarded patient with a constitutional chromosome abnormality, 46,XX,r(21), and acute lymphoblastic leukemia. Scanning electron and differential interference contrast microscopy showed microspikes on the uropodia, but little evidence of cellular motility, despite formation and disappearance of individual uropodia in cell suspensions. The cells rosetted with sheep erythrocytes, suggesting T-cell origin. Cells derived from the bone marrow (80% HMC) showed a high degree of polyploidy (60%) and a bimodal chromosome number of 49 (49,XX,+10,-21, +3 rings) and 94 (6 no. 10, 3 no. 18, 2 no. 21 chromosomes, 3 ring chromosomes, plus 4 copies of each other chromosome).

Bone Marrow↗

Electrocardiographic exercise test in patients with abnormal T waves at rest.

The results of submaximal ECG exercise test were evaluated in six leads recorded simultaneously in two groups of patients with T-wave abnormalities in one or more of Leads I, II, and V4-6. Group I included 185 patients with documented ischemic heart disease (IHD) and Group II 28 patients in whom IHD appeared unlikely. The test was positive in 88 per cent of patients in Group I and in 4 per cent of patients in Group II. In the majority of patients in both groups the T wave either did not change or became more positive or less negative after exercise. The pattern of exercise-induced T-wave changes was similar in patients with and without IHD, and was influenced predominantly by the physiologic effects of exercise. T-wave normalization after exercise occurred frequently in patients with and without IHD, and in patients with positive and negative exercise tests. Our results suggest that T-wave abnormalities, not caused by hypertrophy, conduction disturbances, drugs, or electrolyte imbalance, do not modify the results of submaximal ECG stress test, and that behavior of T wave after exercise does not alter the interpretation of the postexercise ECG. The independent behavior of the S-T segment and T wave after exercise in consistent with the theory that the S-T segment and the T wave are generated by different components of the ventricular action potential.

Adult↗

Phase-randomized laser illumination for microscopy.

A simple apparatus is described that phase randomizes the output of a continuous argon ion laser, so that it may be used as a source of high intensity, monochromatic light for microscopy. The phase-randomizing device can be used with any laser, polarized or unpolarized, and of any desired power output and wavelength, including dye lasers for spectral studies. The randomizing system can be adapted to any light microscope and any optical system including bright-field, phase-contrast, Nomarski differential-interference, dark-field, and split-beam interference systems such as the Jamin-Lebedeff System. It can be used for a variety of photometric and photographic studies. The 514-nm wavelength appears to be relatively harmless to a variety of cells.

Lasers↗

Behaviour of kinetochore fibres in Haemanthus katherinae during anaphase movements of chromosomes.

A laser light source along with a new method of preparing endosperm cells of Haemanthus katherinae for differential interference contrast (DIC) microscopy has led to increased visibility of kinetochore fibres. Little information is available concerning the behaviour of these fibres during anaphase in living cells. In metaphase, kinetochore fibres are seen as distinct bundles of microtubules, here referred to as 'filaments', extending from the kinetochore to the 'diffuse' pole. They possess an apparent globular substructure which corresponds to the moving 'particles or states' described previously from ciné films. In early anaphase, the filaments of each kinetochore fibre lose their parallel orientation characteristic of metaphase and splay out so that the more peripheral filaments intermingle with those of other kinetochore fibres. This process begins at the poles and proceeds as a wave toward the kinetochores as chromosomal movement progresses. This behaviour has been examined in relation to a number of proposed models for the mechanism of chromosome movement and has been found to place some constraints on some models but to be consistent with any model that hypothesizes that chromosomes move as a consequence of cumulative cohesive lateral interactions of microtubules.

Anaphase↗

Membrane behavior of exocytic vesicles. II. Fate of the trichocyst membranes in Paramecium after induced trichocyst discharge.

A specific exocytic process, the discharge of spindle trichocyts of Paramecium caudatum, was examined by means of the electron microscope. This exocytosis is induced by an electric shock simultaneously in nearly all of the trichocysts (ca. 6,000-8,000) of a single cell. Single paramecia were subjected to the shock and then fixed at defined times after the shock so that the temporal sequence of the pattern of changes of the trichocyst membranes after exocytosis could be studied. The trichocyst vacuoles fuse with the plasma membrane only for the length of time required for expulsion to take place. After exocytosis, the membrane of the vacuole does not become incorporated into the plasma membrane; rather, the collapsed vacuole is pinched off and breaks up within the cytoplasm. The membrane vesiculates into small units which can no longer be distinguished from vesicles of the same dimensions that exist normally within the cell's cytoplasm. The entire process is completed within 5-10 min. These results differ from the incorporation of mucocyst membranes into the plasma membrane as proposed for Tetrahymena.

Animals↗

Painless ST-segment depression in patients with angina pectoris. Correlation with daily activities and cigarette smoking.

Ambulatory electrocardiographic monitoring was employed in 33 patients with angina pectoris and abnormal stress tests to determine the frequency with which myocardial ischemia manifested by painless ST-segment depression occurred during normal activity. ST-segment depression occurred in 24 patients during the monitoring period; and in 21, it occurred either solely in the absence of pain or both with and without pain. Of 109 recorded episodes of ST-segment depression, 61 percent were painless. The frequency of painless ST-segment depression was independent of activity other than automobile driving, during which all episodes were painless. In patients who smoked cigarettes, ST-segment depression was more common while smoking, but the incidence of painless ST-segment depression was not altered. The study indicates that ST-segment depression occurs more commonly in the absence than in the presence of chest pain and that ambulatory electrocardiographic monitoring is a useful method of determining the frequency of myocardial ischemia during normal daily activity.

Adult↗

Evidence for firm linkages between microtubules and membrane-bounded vesicles.

Direct evidence is presented in support of the widely held idea that membrane-bounded vesicles can bind firmly to microtubules. This is shown in P. caudatum which contains ribbons of straight microtubules located in open cytoplasm and uniquely associated with the disk-shaped vesicles. These vesicles frequently lie flat against the face of the ribbons at a constant distance of 30-40 nm. Under certain conditions the ribbons are compressed into zigzag pattern, but the vesicles continue to maintain their 30-40 nm spacing with the tubules and The author's interpretation of this phenomena is that the vesicles and the microtubules are strongly bound together. This interaction appears to be via a filamentous material rather than bridges.

Animals↗

Cytochemical localization of peroxisomes in Tetrahymena pyriformis.

Pronounced staining of Tetrahymena pyriformis peroxisomes was demonstrated when glutaraldehyde-fixed cells were incubated in an alkaline medium containing 3,3'-diaminobenzidine and hydrogen peroxide. A variable amount of electron-opague deposit was observed when cells were incubated in diaminobenzidine and H2O2 for 1 hr while an intense deposit followed incubation for 4 hr in the same medium. The staining was abolished completely when 3-amino-1,2,4-triazole, KCN or NaN3 was added to the incubation medium. Based on these cytochemical observations and the morphologic identification by size, shape and other ultrastructural details, it is suggested that this study presents evidence for a conclusive morphologic identification of Tetrahymena peroxisomes.

Amitrole↗