Biomedical subjects
Publications and source records attributed to M Adrian.
Numerous papillomata affecting the skin of the dorsal part of the body occur in a population of Lacerta viridis reared in the laboratory. Electron microscopic study of three of these growths revealed the presence of viral particles belonging, from the morphological aspect, to the groups of the viruses of Herpes, Reo and Papova.
Based on characteristics of mitochondria and on the amount of lipid inclusions, a distinction between somitic cells and mesoblastic somatopleural cells is possible, at the early stages of the development of the limb bud in Reptiles (Anguis fragilis and Lacerta viridis). The dislocation of the ventral processes of the somites and the localisation of the somitic cells in the mesoblast of the anterior limb buds could be studied.
Ultrastructural characteristics of the cells of the apical crest, of the mesoblast and of the ventral processes of somites, in the anlage of the anterior limb buds of embryos of the slow-worm (Anguis fragilis) and of the green lizard (Lacerta viridis) are described at early stages of the development. Differences between the two species studied are brought to light.
An electron microscopic study of the components of anterior limb buds of the slow-worm (Anguis fragilis) and of the green lizard (Lacerta viridis) (embryos of Anguis whose allantoic bud reach 0,7 to 4 mm of length; embryos of Lacerta 2 to 7 days old) provides data on the cytological characteristics of the components of the limb bud at these early stages. 1. The cells of the distal extremity of the somitic processes extending in the limb bud of Anguis and Lacerta, are elongated cells with ovoid nuclei containing large nucleolus; they possess mitochondria always thin and with dense matrix; they are rich in lipid droplets; they possess cilia; they are devoid of myofilaments; endoplasmic reticulum, free ribosomes and polyribosomes are abundant. Golgi networks display signs of activity. These characteristics are also observed in the cells of the "dermatome" layer of the dermo-myotome; and so, it appears probable that the cells of the "dermatome". Furthermore, in Anguis embryos, the cells of the distal extremities of the somitic processes possess numerous lysosomes and a certain number of cells among them, degenerate early. 2. The somatopleural mesoblastic cells of the limb bud of Anguis and Lacerta embryos keep the characters of the cells of the mesodermic layer of lateral plate from which they originate; they have rounded nuclei, cilia, and their mitochondria are always larger and more transparent to electrons, than the ones of cells of the somitic processes and of cells of the epiblastic apical crest. Golgi networks are well developped, endoplasmic reticulum is abundant, lipid droplets are rare. 3. The processes of somites which extend in the dorsal part of the limb bud of Anguis embryos are cords of cells with thin lumina; at the stage of the allantoic bud of 0,6 to 0,8 mm long, the distal extremity of these processes dislocate in group of cells which afterwards dissociate, releasing individual somitic cells which are integrated among the mesoblastic somatopleural cells. In young lizard embryos (2 to 4 days old) the distal extremity of the somitic processes enlarges into a vesicle from which cells are released and penetrate in the mesoblast of the limb bud. 4. The somitic cells released from the somitic processes of Anguis and Lacerta keep--at least at early stages--the cytological characteristics they displayed when they were still in situ in the somitic processes: grounded on the presence or absence of lipid droplets, on the width and density of the mitochondria, the distinction, at these stages, between the somitic and mesoblastic somatoplerual cells is possible; and it is also possible to observe the integration of the somitic cells into the mesoblast. This study brings the demonstration of the cellular contribution of the somites to the formation of the limb bud in Reptiles. 5...
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Isolated rat hearts were perfused according to Langendorff and as a working heart preparation with glucose as the only exogenous substrate under nonrecirculating conditions to avoid accumulation of heart metabolites and, thereby, changes in the composition of the perfusion medium. In the absence of insulin or at low work, oxidation of endogenous substrates as glycogen is of importance for myocardial energy metabolism. Accordingly, about 1/3 of the glucose oxidized by the heart was derived from myocardial glycogen. Lipolysis of endogenous triglycerides and oxidation of the fatty acids produced were, however, low in normal rat hearts. By contrast, in the presence of insulin or at high work load endogenous substrates play a minor role for energy provision. About 80% of the total oxygen consumption could be attributed to the oxidation of exogenous glucose. Furthermore, insulin exerted its major effect in accelerating glucose uptake and glycolysis, but had little influence on PDH-activity. Insulin increased lipolysis in control hearts, however, changes in the endogenous triglycerides were less than valves calculated from the rate of lipolysis. Thus, glycerol release can be taken as a measure for lipolysis, but not as a measure for fatty acid oxidation, since the produced fatty acids were partly reesterified to glycerides. On the basis of the metabolic data obtained, the oxygen and energy balance was calculated. We conclude that a sufficient energy provision is only warranted if the rat heart is perfused either in the presence of insulin or at higher--more physiological--work load.
The scanning probe microscopies applied to the sequencing of DNA is a challenging goal attempted by several groups. But one limitant parameter has been the sample preparation of DNA molecules. Here we report how to hold DNA molecules fixed on mica substrate and we show the three-dimensional configuration of double-stranded DNA obtained with our scanning force microscope. We can image DNA under negative supercoiling, a feature of general importance controlling the activities of DNA. We compared the electron micrographs of a carbon replica of the same DNA specimen with scanning force images which demonstrates well the feasibility and accuracy of our scanning probe measurements.
A procedure is presented for the preparation of thin layers of vitrified biological suspensions in the presence of ammonium molybdate, which we term cryo-negative staining. The direct blotting of sample plus stain solution on holey carbon supports produces thin aqueous films across the holes, which are routinely thinner than the aqueous film produced by conventional negative staining on a continuous carbon layer. Because of this, a higher than usual concentration of negative stain (ca. 16% rather than 2%) is required for cryo-negative staining in order to produce an optimal image contrast. The maintenance of the hydrated state, the absence of adsorption to a carbon film and associated sample flattening, together with reduced stain granularity, generates high contrast cryo-images of superior quality to conventional air-dry negative staining. Image features characteristic of unstained vitrified cryo-electron microscopic specimens are present, but with reverse contrast. Examples of cryo-negative staining of several particulate biological samples are shown, including bacteriophage T2, tobacco mosaic virus (TMV), bovine liver catalase crystals, tomato bushy stunt virus (TBSV), turnip yellow mosaic virus (TYMV), keyhole limpet hemocyanin (KLH) types 1 and 2, the 20S proteasome from moss and the E. coli chaperone GroEL. Densitometric quantitation of the mass-density of cryo-negatively stained bacteriophage T2 specimens before and after freeze-drying within the TEM indicates a water content of 30% in the vitreous specimen. Determination of the image resolution from cryo-negatively stained TMV rods and catalase crystals shows the presence of optical diffraction data to ca. 10 A and 11.5 A, respectively. For cryo-negatively stained vitrified catalase crystals, electron diffraction shows that atomic resolution is preserved (to better than 20 diffraction orders and less than 3 A). The electron diffraction resolution is reduced to ca. 10 A when catalase crystal specimens are prepared without freezing or when they are freeze-dried in the electron microscope. Thin vitrified films of TMV, TBSV and TYMV in the presence of 16% ammonium molybdate show a clear indication of two-dimensional (2-D) order, confirmed by single particle orientational analysis of TBSV and 2-D crystallographic analysis of TYMV. These observations are in accord with earlier claims that ammonium molybdate induces 2-D array and crystal formation from viruses and macromolecules during drying onto mica. Three-dimensional analysis of the TBSV sample using the tools of icosahedral reconstruction revealed that a significant fraction of the particles were distorted. A reconstruction from a subset of undistorted particles produced the characteristic T = 3 dimer clustered structure of TBSV, although the spikes are shortened relative to the structure defined by X-ray crystallography. The 20S proteasome, GroEL, catalase, bacteriophage T2, TMV, TBSV and TYMV all show no indication of sample instability during cryo-negative staining. However, detectable dissociation of the KLH2 oligomers in the presence of the high concentration of ammonium molybdate conforms with existing knowledge on the molybdate-induced dissociation of this molecule. This indicates that the possibility of sample-stain interaction in solution, prior to vitrification, must always be carefully assessed.
Thin vitrified layers of unfixed, unstained and unsupported virus suspensions can be prepared for observation by cryo-electron microscopy in easily controlled conditions. The viral particles appear free from the kind of damage caused by dehydration, freezing or adsorption to a support that is encountered in preparing biological samples for conventional electron microscopy. Cryo-electron microscopy of vitrified specimens offers possibilities for high resolution observations that compare favourably with any other electron microscopical method.
The basic principles of the architecture of many viral protein shells have been successfully established from electron microscopy and X-ray data, but enveloped viruses have been more difficult to study because they resist crystallization and are easily deformed when prepared for electron microscopy. To avoid the limitations of conventional techniques when applied to enveloped viruses, we have used a cryo-electron microscopy method in which unfixed and unstained viruses are observed in an unsupported thin layer of vitrified suspension. Because of electron beam damage, the many different views required for high-resolution three-dimensional reconstruction cannot be obtained from a tilt series of the same particle. The images of many differently oriented viruses are combined using a novel reconstruction method, 'reconstruction by optimized series expansion' (ROSE). The structure of the envelope of Semliki Forest virus has been reconstructed to 3.5-nm resolution. The T = 4 geometry of the surface lattice, the shape of the trimeric spikes and their arrangement on the lipid bilayer are visualized.
Multiple regression analysis of cross-sectional 1985-1986 Ontario county data indicated that the presence of Native Indians on reserves is a significant factor in explaining differences in county alcohol consumption levels. Consumption in counties with reserves was higher than in those without reserves by roughly 1.48 liters of absolute alcohol per adult; consumption increased as the Native reserve population increased (p less than 0.05). When income, employment, household crowding, type of industrial activity, northern isolation, and tourism were included, we could account for over 60% of the variation in alcohol consumption between Ontario counties (p less than 0.01). Every extra $1,000 in income per tax return was associated with a 0.297-liter reduction in absolute alcohol consumption. Efforts to reduce alcohol consumption in the Native population would have their greatest impact when associated with improved economic conditions.
This paper reviews intercultural variability of substance use behaviors, including availability of international statistics on consumption of alcohol and other drugs, as well as the use of drugs available locally only. Within a conceptual framework of intercultural relations, it considers the history of transcultural spread of substance use behaviors and possible reactions to the introduction of new drugs within a culture or jurisdiction, including illustrations of the "law of alien poisons." Although intercultural views of substance use have generally concentrated on majority groups' views of substance use in minority groups, minority and non-Western views of substance use need to be considered in the context of increasing international and intercultural communications that increase the rate at which substance use behaviors spread. Both Western and non-Western experiences with substance use and misuse must be taken into account so that better interventions can be developed to deal with addictions and other substance-related problems.
PURPOSE: The goal of this study was to develop a method to measure the impact and cost-effectiveness of health promotion. DESIGN: Age- and sex-specific changes in life expectancy in Canada between 1970 to 1972 and 1985 to 1987, after the introduction of national health insurance (1970) and health promotion (1975), are used to assess the impact due to biological hardiness, improvements in the health care field, and the effects of health promotion. SUBJECTS: The subjects were the total male and female population of Canada between the years 1970 to 1972 and 1985 to 1987. MEASURES: Life expectancy by years of age by sex was the measurement used. RESULTS: A method is presented that allows the calculation of the differential effects of health promotion, health care, and biological hardiness on changes in life expectancy based on sequential subtraction of life expectancies for one-year age cohorts over a 15-year period. Results were obtained for each year of age for men and women, showing gender and age differences in the relative impact of the three factors. In this illustrative example using Canadian data, health promotion was found to have less impact on longevity than health care or biological hardiness. However, of the three, health promotion was the most cost-effective. CONCLUSION: This method can be used to quantify changes in life span due to health promotion, health care, and biological hardiness for men and women at each year of age and to relate this to health expenditures for the whole population. The method is limited in that it cannot determine the relative impact of other factors that can affect life expectancy such as environmental changes or social trends.
Carrying on their study about the enzymatic activity of the otospongiotic micro-foci and the hydrolytic activity of the perilymph, the authors present their work concerning the value of trypsin and alpha-1-anti-trypsin in the perilymph of otospongiotic patients operated on by stapedectomy. They describe the method used and give the obtained results which permit to believe that the values of trypsin, and a contrario of anti-trypsin, appear to constitute an index of the severity of the progression. Moreover, the authors have investigated the eventual toxic action of various trypsin concentrations on the hair cells of the Corti organ in the guinea-pig. This study has been carried through successfully by means of the electron microscopic scanning method which allows to obtain an "electron cochleogramme in situ". They explain the method and come to the conclusion that the intensity and extension of the Corti hair cells alterations in a cochlea receiving a perfusion with trypsin solutions, are tightly related to the trypsin concentration. However, they do not pretend so far that these results reflect exactly the anatomo-pathologic reality during the slow progression of otospongiosis towards cochlear deterioration. These two series of experimentation appear to confirm their enzymatic concept of otospongiosis: the long, slow and extremely capricious progression of the disease is the duplicate of its cytologic progression, the contact between hydroltic enzymes and Corti cells having the same capricious character. These findings perfectly corroborate the previus ones they made in this field, as well as the correlations they have established between hydrolytic activity of the perilymph and audiometric progression. Consequently, they believe that the mechanism of the otospongiotic disease is provoked by the rupture of the balance trypsin/anti-trypsin in the otospongiotic micro-foci and thus in the inner ear fluids.
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