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  • Articles: DFG German National Licenses  (51,200)
  • 1970-1974  (27,320)
  • 1965-1969  (23,880)
  • Chemistry  (51,012)
  • Bone
  • Nuclear reactions
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  • Articles: DFG German National Licenses  (51,200)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 14 (1974), S. 309-315 
    ISSN: 1432-0827
    Keywords: Osteoblast ; Matrix ; Bone ; Scanning electron microscope
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les territoires sécrétoires des ostéoblastes d'os pariétal de rats sont déterminées en utilisant la microscopie électronique à balayage. Le territoire moyen de 4.620 cellules, dans 19 territoires, est de 154 μm2 par ostéoblaste. Les valeurs extrêmes par champ varient de 136 à 177 μm2 par ostéoblaste. Quatre cent cellules sont mesurées individuellement; la valeur moyenne par ostéoblaste est de 143 μm3 avec une déviation standard de 33. Le taux d'apposition journalier, mesuré par la tétracycline pendant 8 jours, est de 3.12 μm (déviation standard 0.22). Ce qui correspond à une production matricielle journalière d'environ 470 μm3 par ostéoblaste.
    Abstract: Zusammenfassung Die Ausscheidungsbereiche von Ratten-Osteoblasten des Scheitelbeines wurden mit dem Raster-Elektronenmikroskop direkt gemessen. Der durchschnittliche Bereich von 4620 Zellen in 19 Gesichtsfeldern war 154 μm2 per osteoblast. Der Streubereich lag in den verschiedenen Gesichtsfeldern zwischen 136 und 177 μm2 per Osteoblast. 400 Zellen wurden einzeln gemessen. Bei diesen war der Durchschnittswert per Osteoblast 143 μm2, mit einer Standard-Abweichung von 33. Die tägliche Anlagerungsrate während einer Periode von 8 Tagen war 3,12 μm (Standard-Abweichung 0,22); sie wurde mittels Tetracyclinmarkierung der Mineralisierungsfront gemessen. Dies ergab eine tägliche Produktionsrate der Matrix von etwa 470 μm3 per Osteoblast.
    Notes: Abstract The secretory territories of rat osteoblasts on the parietal bone were measured directly using scanning electron microscopy. The mean territory of 4620 cells in 19 fields was 154 μm2 per osteoblast. The range for the fields was 136 to 177 μm2 per osteoblast. Four hundred cells were measured individually—for these the mean value per osteoblast was 143 μm2 with a standard deviation of 33. The daily rate of apposition over an 8 day period was 3.12 μm (standard deviation 0.22) measured by tetracycline marking of the mineral front. This gave a daily matrix production rate of approximately 470 μm3 per osteoblast.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 14 (1974), S. 327-331 
    ISSN: 1432-0827
    Keywords: Bone ; Piezoelectricity ; Collagen ; Aging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The piezoelectric constant of mature and immature bone (defined herein) has been measured in an effort to determine whether it varies with age. It was found that the average value of the piezoelectric constantd 14 of femur from three week old calves was 58% of the value of femur from three year old bulls. The results were interpreted to indicate qualitative differences in the corresponding collagen matrices. Mature human tibia from males ranging in age from 21 to 53 years of age showed a small but significant increase ind 14 with age. Some data concerning diseased human bone, and well-preserved human bone excavated in Peru are also presented.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 14 (1974), S. 333-337 
    ISSN: 1432-0827
    Keywords: Bone ; Materials ; Histology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des métaux poreux et des céramiques sont implantés dans l'osin vivo et sont prélevés, après différents intervalles, pour étudier histologiquement la colonisation tissulaire. Les implants prélevés sont fixés avec les tissus environnants et inclus dans des epoxy-résines, de faible viscosité. Les échantillons sont coupés à l'aide d'une scie diamantée et fixés sur des lames pétrographiques, pour être ensuite amincis à environ 75 μm. Ils sont légèrement colorés au bleu de méthylène et à l'alizarine rouge S, qui donne un bon contraste entre tissus durs et mous et démontrent la présence d'os calcifié dans les implants poreux métalliques et céramiques.
    Abstract: Zusammenfassung Poröse Metall- und Keramikstücke wurden in vivo in Knochen implantiert und nach verschiedenen Intervallen entnommen, um histologisch zu untersuchen, wie weit sie ins Gewebe eingewachsen waren. Die entnommenen Implantate mit dem sie umgebenden Gewebe wurden fixiert, dehydriert und in ein niedrigvisköses Epoxy-Harz eingebettet. Die Proben wurden mit einer Diamantsäge in dicke Schnitte zerteilt, auf Steinscheiben zementiert und von Hand auf etwa 75 μm geschliffen. Sie wurden mit Methylenblau und Alizarin-Rot S schwach gefärbt; dies ergibt einen großen Kontrast zwischen Knochen und Weichgewebe und zeigt die Bildung von verkalktem Knochen innerhalb poröser Metall- und Keramik-Implantate deutlich.
    Notes: Abstract Porous metals and ceramics were implanted in bonein vivo and retrieved at intervals for histological evaluation of tissue ingrowth. Recovered implants with surrounding tissues were fixed, dehydrated and embedded in a low-viscosity epoxy resin embedding medium. The embedded samples were cut into thick sections with a diamond saw, cemented to petrographic slides, and hand-ground to approximately 75 μm. They were lightly stained with methylene blue and Alizarin Red S, which provided great contrast between the bone and soft tissues, and clearly demonstrated the formation of calcified bone within porous metal and ceramic implants.
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  • 4
    ISSN: 1432-0827
    Keywords: Osteogenesis ; Osteoinduction ; Bone ; Matrix ; Cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Bone matrix demineralized in 0.6 N HCl at 2° for 24 h and implanted in muscle in allogeneic rats possesses consistently reproducible bone morphogenetic activity. Experiments on implants of matrix, obtained from donors injected with3H-tyrosine or3H-tryptophan, or Na35SO4, suggest that bone morphogenetic property is a protein or apart of a protein that is (1) insoluble in buffer solutions, pH 3.6 and 5.0; (2) degraded in buffer solutions at pH 7.4 by an endogenous sulfhydryl-group neutral proteinase; (3) digested by trypsin at 15° within 8 h without solubilization of the helical regions, possibly even without degradation of the nonhelical ends of the bone collagen molecule, and without any loss of the periodic ultrastructure of the collagen fibrils; (4) degraded or removed by 0.1 N NaOH at 2° within 24 h without solubilization of collagen; (5) biologically active even after nitration of tyrosyl groups with tetranitromethane. The release of only one-third of the radioactivity with loss of nearly all yield of new bone by limited tryptic digestion of3H-borohydride-reduced matrix indicates that the bone morphogenetic response is the function of a non-collagenous component. Autoradiographs of implants of matrix with non-collagenous proteins labelled with3H-tryptophan,3H-tyrosine, or both3H-tyrosine and3H-phenyl-alanine demonstrate random dissemination of the radioactive constituents and no evidence of local transfer of labelled proteins or soluble protein derivatives. Hypothetically, the bone morphogenetic response is controlled by an insoluble acidic bone morphogenetic protein or polypeptide (BMP) and a soluble neutral proteinase (BMP-ase) resembling trypsin in activity except functionally more specific for BMP. Firmly bound but separable from bone collagen, BMP is one of many short-lived morphogenetic substances appearing and disappearing throughout embryonic development and persisting in postfetal life. Where the BMP receptor resides and how it activates cell mechanisms of differential repression and derepression of such genes as code for osteogenesis is unknown.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 15 (1974), S. 333-339 
    ISSN: 1432-0827
    Keywords: Vitamin D ; Bone ; Resorption ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract A series of analogues of vitamin D have been tested for their ability to stimulate bone resorption in two test systems used previously to investigate the metabolites of vitamine D. These analogues were tested (a) by directly comparing their action on bone explants of mouse half-calvariain vitro, and (b) by injecting them into young mice and measuring the degree of resorptionin vitro when explants were prepared 18 hours after the injection. It is concluded that the key functional groups concerned with enhancing the activity of vitamin D3 are the 1α- and the 25-hydroxyl,both together; the cis ring structure for ring A appears necessary. 1α-Hydroxycholecalciferol (1α-OHD3) is about as active as 25-OHD3 in the direct test, but its potency is much nearer to that of 1,25-(OH)2D3 when tested by the second (indirect) method; it seems likely that 1α-OHD3 is converted into 1,25-(OH)2D3 in vivo. The results are discussed in relation to the designing of analogues for clinical and experimental use.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 16 (1974), S. 145-152 
    ISSN: 1432-0827
    Keywords: EHDP ; Bone ; Chemistry ; Serum ; Rabbits
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The effects of disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) on bone and serum chemistry were investigated in adult rabbits. EHDP was administered by subcutaneous injection at doses of 0.25, 2.5 and 10 mg/kg body weight/day for of 28 days. Blood samples were obtained weekly from each rabbit and serum levels of total calcium, ionized calcium, inorganic phosphate and alkaline phosphatase were determined. At the end of the treatment period all rabbits were sacrificed and the tibiae removed for chemical analysis and histological evaluation. The effect of EHDP administration on serum chemistry was both dose- and time-related. The highest of the three doses, 10 mg/kg/day, resulted in a time-related decrease in total serum calcium. This dose also caused a rapid but transient reduction in serum ionized calcium. The effect of EHDP on serum inorganic phosphate was biphasic. Administration of 2.5 mg/kg/day resulted in a time-related elevation in this parameter, whereas the 10 mg/kg/day dose resulted in a time-related hypophosphatemic response. There were no significant drug-related changes in tibial fat-free dry weight, ash weight, total calcium or total phosphorus values. However, administration of 2.5 and 10 mg/kg/day EHDP resulted in increased osteoid tissue as measured histologically. These results are compared with data from other EHDP studies, and discussed in relation to the maturity and growth-state of the experimental animals.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 16 (1974), S. 261-275 
    ISSN: 1432-0827
    Keywords: Dentine ; Bone ; Mineralization ; Density gradient fractionation ; Proteoglycans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract In order to obtain enough material to analyse the organic matrix of mineralizing tissue a technique for preparative separation according to the degree of mineralization was developed. The method employs ultracentrifugation of powdered material in density gradients made from acetone and bromoform. The density range of the fractions is checked by refractive index measurements of the gradient medium. The amount of the material in the fractions is checked by weighing and their degree of mineralization is estimated by determining the Ca/N and P/N ratios. The homogeneity of the fractions is determined by soft X-ray microscopy. Isolated dissected microscopic bone structure (osteones and lamellar bone fragments) with different degrees of mineralization were fractionated in this way. Chromatography on Sepharose 2B of proteoglycans from costal cartilage exposed to an acetone-bromoform gradient revealed no effect of the gradient medium on the molecular size of the proteoglycans.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 14 (1974), S. 153-160 
    ISSN: 1432-0827
    Keywords: Lysosomes ; Bone ; Resorption ; Osteoclast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des techniques histochimiques de microscope photonique et électronique montrent que les ostéoclastes métaphysaires chez des rats “sans incisive” présentent des quantités plus importantes de phosphatase acide, d'acryl sulfatase et de trimétaphosphatase acide lysosomiales. L'activité en phosphatase lysosomiale à pH neutre est aussi plus élevée dans les ostéoclastes métaphysaires, sauf dans les cellules situées sous la métaphyse, où l'enzyme est absente. L'absence de résorption de la matrice organique semble en rapport avec l'absence de bordures en brosse et une absence d'enzyme lysosomiale extracellulaire. Malgré cette absence, une dissolution de cristaux inorganiques a été mise en évidence au microscope électronique, suggérant que la dissolution minérale est un processus distinct de la désintégration enzymatique de la matrice organique.
    Abstract: Zusammenfassung Histochemische Techniken für Licht- und Elektronenmikroskopie zeigten, daß metaphysäre Osteoklasten “Schneidezahl-loser” (SL) Ratten erhöhte Mengen von lysosomaler saurer Phosphatase, von Arylsulfatase und von saurer Trimetaphosphatase enthielten. Die Aktivität der lysosomalen Phosphatase bei neutralem pH war in den metaphysären Osteoklasten ebenfalls erhöht, außer in den Zellen direkt unterhalb der Wachstumsplatte, wo dieses Enzym nicht vorkam. Es konnte überhaupt keine Resorption der organischen Matrix festgestellt werden, was übereinzustimmen schien mit der Abwesenheit eines gekräuselten Saumes und einer gleichzeitigen Abwesenheit des extrazellulären lysosomalen Enzyms. Trotzdem wurde im Elektronenmikroskop das Verschwinden anorganischer Kristalle festgestellt, was darauf schließen läßt, daß die Auflösung des Minerals nicht derselbe Vorgang ist wie die enzymatische Auflösung der organischen Matrix.
    Notes: Abstract Histochemical techniques for light and electron microscopy showed that metaphyseal osteoclasts in “incisors absent” rats contained greater than normal amounts of lysosomal acid phosphatase, aryl sulfatase and acid trimetaphosphatase. Lysosomal phosphatase activity at neutral pH was also elevated in the metaphyseal osteoclasts except in those cells immediately beneath the growth plate, where this enzyme was absent. The failure of any discernable resorption of organic matrix appeared to correlate with the absence of a ruffled border and a concomittant absence of extracellular lysosomal enzyme. Despite this failure, electron microscopic evidence of inorganic crystal removal was noted, suggesting that mineral dissolution represents a separate process from the enzymatic breakdown of organic matrix.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 16 (1974), S. 13-36 
    ISSN: 1432-0827
    Keywords: Osteoclast ; Bone ; Resorption ; Crystals ; Organic-inorganic relationships
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The organic-inorganic relationships in bone matrix undergoing osteoclastic resorption have been studied in rat tibial diaphyses using electron microscope techniques in an attempt to identify the steps of the resorption process. Results suggest that bone resorption occurs in two phases: the first, an extracellular phase, leads to bone matrix fragmentation and partial dissolution, and the second, an intracellular phase, to complete digestion of the breakdown products of the bone matrix. The first component of the bone matrix to be attacked by the osteoclast is the ground substance. This induces the release of the crystals lying between, and on, the collagen fibrils; any crystals lying within fibrils are released later, when the fibrils break up. As this stage proceeds, the collagen fibrils retain their normal intrinsic texture, but gradually loose their lateral aggregation, appearing as individual fibrils (some of them uncovered by crystals), mixed with fragments of fibrils and many free crystals. The loosened but otherwise structurally normal collagen fibrils, and their fragments, are strongly argyrophilic. Complete dissolution of the disaggregated fibrils occurs outside the cell, both in the resorption zone and in the initial portion of the channels of the ruffled border. The free crystals present in the resorption zone and those phagocytosed in cytoplasmic vacuoles are organic-inorganic structures, whose organic component (the crystal ghost) is, at least in part, of proteoglycan nature. Dissolution of inorganic material occurs within the cytoplasmic vacuoles of the osteoclasts. Results are viewed in relation to the process of bone resorption and, as far as crystal ghosts are concerned, to that of bone calcification. A tentative summary of the various steps involved in the mechanism of bone resorption is given.
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  • 10
    ISSN: 1432-0827
    Keywords: Bone ; Parathyroid ; 25-Hydroxycholecalciferol ; Adenosine 3′, 5′ cyclic monophosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Phenytoin, at a concentration which is known to inhibit parathyroid extract-induced bone resorptionin vitro, has been shown to have no significant effect on either the cyclic AMP content of four-day-old mouse calvaria which had been in tissue culture for 48 h or on the increase in cyclic AMP content occurring after the addition of parathyroid extract to the medium. Phenytoin has also been shown to be an effective inhibitor of bone resorptionin vitro induced by 25-hydroxycholecalciferol. These observations suggest that the inhibitory effect of phenytoin on parathyroid extract and hormone-induced bone resorption takes place beyond the cyclic AMP stage.
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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 16 (1974), S. 219-226 
    ISSN: 1432-0827
    Keywords: Bone ; Phosphate ; ATP ; Calcium ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract An active constituent of the leaves ofSolanum malacoxylon (SM) is shown to promote calcium resorption and citrate production in embryonic chick frontal bone culturedin vitro. When injected into rats, SM reduces the ATP content of liver and kidneys. This phenomenon may be related to the ability of SM to stimulate mitochondrial ATPase activity at pH 9.4. SM significantly reduces the concentration of phosphate necessary to alter the uptake of calcium and respiration of siolated mitochondria. The data suggest that SM influences calcium and phosphate metabolism by affecting ion movements into and out of mitochondria.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 143 (1974), S. 159-183 
    ISSN: 1432-0568
    Keywords: Bone ; Femur ; Development ; Gravity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The significance of similitude principles for the postnatal development of the cross-sectional area at midlength of the human and rat femur has been subjected to an investigation based both on theory and on observation. Cross sections were taken from 69 femora of female Sprague-Dawley rats 34, 72, 102, and 840 days old and from the right human femora of 94 males and 58 females ranging from 1 day to 89 years of age. The regression of the square-root of the cross-sectional area divided by π on femur length is linear in the rats. The empirical regression line estimated by the method of least squares from the measurements and the theoretical curve calculated on the assumption of adaptive allometric growth and centrically applied loads are more or less alike in slope and elevation. The corresponding empirical relationship in the human femora turned out statistically nonlinear in the males and linear in the females. Comparing the empirical regression functions with five theoretical curves, it has been revealed that the observations are best approximated by two curves calculated on the assumption of centrically or eccentrically applied loads and adaptive allometric growth of the cross-sectional diameter. It was further uncovered that the cross-sectional diameters at midlength of the femur are not changed isometrically with femur length both in rats and in man. Considering a simple model of the human body in the stance phase of slow walking gait on the scale of a neonate and an adult male, the hypothesis was advanced that the stresses at midlength of the femur are not altered much while the femur grows from neonate to adult size, because the outer and inner radii of the femur actually change during growth in such a way that the effects due to scale are appropriately compensated.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 14 (1974), S. 251-255 
    ISSN: 1432-0827
    Keywords: Cartilage ; Bone ; Vitamin A ; Phosphatases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Au cours d'une étude du cartilage épiphysaire de poulets normaux et déficients en vitamine A, la composition organique et l'activité phosphatasique du cartilage au repos, du cartilage en voie d'ossification et de l'os nouvellement formé ont été mesurées. Le cartilage en voie d'ossification et l'os nouvellement formé ont un contenu plus élevé en matériel inorganique, phosphate et collagène que le cartilage au repos. Une déficience en vitamine A provoque une augmentation du contenu en phospholipide de tous les trois tissus. Le cartilage au repos, en cas de déficience en vitamine A, présente, après homogénéisation et centrifugation, un surnageant dont l'activité en phosphatase alcaline et en glycérophosphatase est plus élevée que celle des témoins. Il semble que les effets enzymatiques de l'avitaminose A soient liés à des altérations de la membrane lysosomiale avec décharge de phosphatases. La minéralisation normale semble aussi faire intervenir une activité phosphatasique.
    Abstract: Zusammenfassung Diese Arbeit beschreibt Untersuchungen des Epiphysenknorpels bei normalen Küken und solchen mit Vitamin-A-Mangel. Die organische Zusammensetzung und die Phosphatase-Aktivität in ruhendem Knorpel, ossifizierendem Knorpel und neugebildetem Knochen wurden gemessen. Der ossifizierende Knorpel und der neugebildete Knochen hatten einen höheren Gehalt an anorganischem Material, an Phosphat und Kollagen als der ruhende Knorpel. Vitamin-A-Mangel führte zu einem erhöhten Phospholipidgehalt in allen drei Geweben. Nach Homogenisierung und Zentrifugierung fand sich im Überstand des ruhenden Knorpels von Vitamin-A-Mangelgewebe eine höhere alkalische Phosphatase- und Glycero-phosphatase-Aktivität als bei den Kontrollen. Es wird angenommen, daß die Wirkung des Vitamin A-Mangels auf die Enzyme im Zusammenhang steht mit der Schädigung der lysosomalen Membran, wobei Phosphatasen freigesetzt werden, und daß die normale Mineralisierung die Phosphatasen-Aktivität ebenfalls anregt.
    Notes: Abstract This paper reports studies of the epiphyseal cartilage in normal and vitamin A deficient chicks. The organic composition and the phosphatase activity in the resting cartilage, ossifying cartilage and new bone were measured. The ossifying cartilage and new bone had a higher content of inorganic material, phosphate and collagen than the resting cartilage. Vitamin A deficiency caused increase in the phospholipid content of all three tissues. The resting cartilage from vitamin A deficient tissue had, after homogenisation and centrifugation, a supernatant with an activity of alkaline phosphatase and glycerophosphatase higher than that in control samples. It is considered that effects of vitamin A deficiency on enzymes are related to defects of the lysosomal membrane with release of phosphatases, and that normal mineralisation also involves phosphatases activity.
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 14 (1974), S. 283-289 
    ISSN: 1432-0827
    Keywords: Sodium fluoride ; Osteoporosis ; Bone ; Strength ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'utilisation thérapeutique de fluorure de sodium a été recommandée dans le traitement de diverses maladies osseuses ostéopéniques. Cet emploi est basé principalement sur les effects ostéosclérotiques bien connus du fluorure de sodium, mais on connait mal son effet sur la force de résistance osseuse. L'influence de diverses concentrations de fluorure de sodium sur la force de résistance osseuse de jeunes rats soumis à des régimes riches et pauvres en calcium a été étudiée. L'administration de fluorure de sodium augmente le diamètre osseux, indiquant une stimulation de la formation de l'os périosté, mais la force de résistance osseuse est réduite ou non modifiée par l'ingestion de fluor.
    Abstract: Zusammenfassung Die therapeutische Verwendung von Natriumfluorid ist für eine Anzahl von Knochenmangel-Krankheiten empfohlen worden. Die Empfehlungen basieren hauptsächlich auf den bekannten osteosklerotischen Wirkungen von Natriumfluorid; über dessen Effekt auf die Knochenstärke ist wenig bekannt. In dieser Arbeit wurde der Einfluß verschiedener Konzentrationen von Natriumfluorid auf die Knochenstärken von wachsenden Ratten mit hoher und niederer Calciumeinnahme untersucht. Die Verabreichung von Natriumfluorid erhöhte den Knochendurchmesser, was auf eine Stimulierung der Periostbildung hinwies, die Knochenstärke wurde jedoch durch Fluorideinnahme herabgesetzt oder nicht beeinflußt.
    Notes: Abstract The therapeutic use of sodium fluoride has been recommended in a variety of osteopenic bone diseases. The recommendations are based mainly on the known osteosclerotic effects of sodium fluoride and little information is available as to its effect on bone strength. The influence of various concentrations of sodium fluoride on bone strength in growing rats on high and low calcium diets was studied. The administration of sodium fluoride increased bone diameter, indicating stimulation of periosteal bone formation, but bone strength was reduced or not affected by fluoride ingestion.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 14 (1974), S. 291-299 
    ISSN: 1432-0827
    Keywords: Bone ; Collagenase ; Lysozyme
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Du lysozyme de blanc d'œuf, ainsi que des protéines basiques, telles que de l'histone et une base dépourvue de protamine, semblent inhiber la collagénase osseuse de souris. L'inhibition de collagénase osseuse de souris par le lysozyme est mise en évidence lorsque l'activité de la collagénase est étudiée en utilisant du collagène comme substrat à l'état solide, mais non lorsque le collagène est utilisé en solution. D'autre part, l'inhibition de l'activité en collagénase par l'histone et une base dépourvue de protamine est observée dans les deux systèmes. L'inhibition de collagénase ossuese de souris par des molécules polyanioniques est intéressante, étant donné que des travaux antérieurs ont montré que plusieurs molécules polyanioniques telles que l'héparine, le sulfate de dextrane et l'acide polyethylenesulfonique augmentent l'activité de collagénase osseuse de souris dans les mêmes conditions expérimentales. Comme le lysozyme est largement présent dans les tissus conjonctifs et que sa concentration carie avec l'ampleur du remaniement tissulaire, l'inhibition de la collagénase osseuse de souris suggère un role éventuel dans la régulation de la dégradation du collagène, pendant le remaniement des tissus collagéniquesin vivo au cours d'états normaux et pathologiques.
    Abstract: Zusammenfassung Lysozyme aus Hühnereiweiß sowie Basis-Proteine, wie z. B. Histon und Protamin-freie Basen, erwiesen sich als Hemmer der Mäuseknochen-Kollagenase. Diese Hemmung durch Lysozym wurde festgestellt, wenn für die Messung der Kollagenase-Aktivität Kollagen in Substanz als Substrat verwendet wurde, nicht aber, wenn das Substrat aus gelöstem Kollagen bestand. Andererseits wurde die Hemmung der Kollagenase-Aktivität durch Histon und Protamin-freie Basen in beiden Versuchssystemen festgestellt. Die Hemmung von Mäuseknochen-Kollagenase durch polykationische Moleküle ist interessant, vor allem im Hinblick auf frühere Feststellungen, daß mehrere polyanionische Moleküle, wie Heparin, Dextransulfat und Polyaethylensulfonsäure, die Aktivität der Mäuseknochen-Kollagenase unter denselben experimentellen Bedingungen erhöhen. Da Lysozym in Bindegeweben überall verteilt ist und da dessen Konzentration mit der Neubildungsrate des Gewebes variiert, läßt die Hemmung der Mäuseknochen-Kollagenase durch Lysozym dessen mögliche Rolle in der Regulierung des Kollagenabbaues vermuten, und zwar während der Neubildung von Kollagengeweben in vivo in normalen und pathologischen Zuständen.
    Notes: Abstract Egg white lysozyme, as well as basic proteins such as histone and protamine-free base, were found to inhibit mouse bone collagenase. The inhibition of mouse bone collagenase by lysozyme was detected when the activity of the collagenase was assayed using collagen as the substrate in the solid state, but not when the collagenase activity was assayed using collagen in solution as the substrate. On the other hand, the inhibition of collagenase activity by histone and protamine-free base was observed in both assay systems. The inhibition of mouse bone collagenase by polycationic molecules is interesting in light of previous findings that several polyanionic molecules, such as heparin, dextran sulfate and polyethylenesulphonic acid, enhance the activity of mouse bone collagenase under the same experimental conditions. Since lysozyme is widely distributed in connective tissues and its concentration varies with the rate of tissue remodeling, the inhibition of mouse bone collagenase by lysozyme suggests its possible role in the regulation of collagen degradation during the remodeling of collagenous tissuesin vivo during normal and pathological states.
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  • 16
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    Calcified tissue international 15 (1974), S. 11-20 
    ISSN: 1432-0827
    Keywords: Bone ; Growth ; Resorption ; Caudal ; Vertebra ; Fluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une méthode de mesure de l'apposition et de la résorption osseuse est mise au point au niveau de la septième vertèbre caudale de jeunes rats, pesant 25–90 g, pendant la période de croissance. Cette pièce osseuse a des avantages certains par rapport à d'autres localisations, car elle est symétrique en coupe transversale et non affectée par des forces corticales. En utilisant trois marqueurs d'os différents, fluorescents en lumière ultra-violette, administrés à des intervalles de 3, 5 ou 7 jours, les modifications de croissance sont observées sur des coupes transversales centrales, non décalcifiées, de la septième vertèbre caudale. Une formation osseuse linéaire se développe à une vitesse moyenne de 10.0 μ/jour et une résorption linéaire de 6.1μ/jour est notée. Pendant la période de croissance observée, la formation osseuse s'observe exclusivement à la surface du périoste et la résorption se fait le long de la surface de l'endoste. A ce niveau l'os est totalement résorbé, chez des animaux de 25 g en 22–26 jours. Il semble que chez des animaux plus âgés, une formation osseuse endostée se développe rendant alors impossible toute étude prolongée dans cette région.
    Abstract: Zusammenfassung Es wird eine Methode zur Messung der Knochenbildungs- und Resorptionsgeschwindigkeiten im siebten Caudalwirbel junger Ratten während ihrer Wachstumsperiode von 25–90 g beschrieben. Dieser Knochen hat gegenüber jenen, die vorgängig untersucht wurden, den großen Vorteil, in transversalen Schnitten symmetrisch und unbeeinflußt von kortikalen Unregelmäßigkeiten zu sein. Die Art der Wachstumsveränderungen wurden an nichtentkalkten zentralen Transversalschnitten der siebten Caudalwirbeln bestimmt; dazu wurden drei verschiedene, farbige, im UVbereich fluoreszierende Knochenmarkierungssubstanzen verabreicht und zwar in 3-, 5- oder 7tägigen Intervallen. Die lineare Knochenbildung fand mit einer mittleren Geschwindigkeit von 10,0 μ pro Tag statt, die lineare Knochenresorption mit einer solchen von 6,1 μ pro Tag. Während der untersuchten Wachstumsperiode fand die Knochenbildung ausschließlich an der Periostoberfläche und die Knochenresportion an der endostalen Oberfläche statt. Der Knochen, der beim 25 g schweren Tier an diesem Ort vorlag, verschwand durch die endostale Resorption innerhalb von 22–26 Tagen
    Notes: Abstract A method for measuring bone formation and resorption rates in the seventh caudal vertebra of young rats during the growth period from 25–90 g is reported. This bone site has unique advantages over those previously studied, being symmetrical in transverse section and uncomplicated by cortical drift. Utilising three different coloured ultraviolet fluorescent bone labelling substances administered at either 3, 5 or 7 day intervals, the nature of growth changes was determined from undecalcified central transverse sections of the seventh caudal vertebrae. Linear bone formation occurred at a mean rate of 10.0 μ/day and linear bone resorption at 6.1 μ/day. During the growth period studied, bone formation occurred exclusively on the periosteal surface and bone resorption on the endosteal surface. Bone existing in this site in 25 g animals was completely removed by endosteal resorption within 22–26 days. Evidence exists that in older rats endosteal bone formation occurs and renders the site unsuitable for long term studies.
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  • 17
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    Calcified tissue international 15 (1974), S. 315-324 
    ISSN: 1432-0827
    Keywords: Bone ; Mineral ; Osteogenesis ; Interference microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract A method based on tetracycline labeling was developed for selecting, in frozen sections of fetal mouse femur, bone foci approximately 6, 24, 48 and 72 h of age. Microinterferometric measurements of these foci permitted a calculation of the effective thickness (t) and refractive index (n) of each focus. After demineralizing the sections by a method which left the organic portion of the bone intact, the foci were re-measured andt andn of the organic compartment of each focus were determined;t andn of the mineral compartment were calculated by difference. A sharp decrease and subsequent rise in then of whole bone occurred between 6 and 48 h. These changes derived from the mineral compartment, and were thought to have resulted from the formation of amorphous calcium phosphate and hydroxyapatite, respectively. The mineral present in 6 h foci, however, was believed to be some precursor of amorphous calcium phosphate.
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  • 18
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    Calcified tissue international 15 (1974), S. 329-332 
    ISSN: 1432-0827
    Keywords: Microradiography ; Bone ; X-ray
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract A low cost, self-contained, commercially available, X-ray apparatus which possesses the essential qualifications necessary to produce high resolution microradiographs of undecalcified bone is described. The widespread current use of this machine in pathology laboratories may make purchase of such a unit optional.
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  • 19
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    Calcified tissue international 16 (1974), S. 73-88 
    ISSN: 1432-0827
    Keywords: Dentine ; Bone ; Histocytochemistry ; Physiological Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Histochemical methods were used to detect differences in amino acid reactive groups between bone and osteoid, dentine and predentine. Three major differences were found: Predentine and osteoid contained more reactive sulphydryls than their mineralized counterparts. The greatest reaction for this reactive group was obtained at the mineralizing front. Dentine and bone contained a complex between amino and carboxylic acid groups; predentine and osteoid did not have this complex and carboxylic acid residues were less in these tissues. Dentine and bone contained much greater amounts of histidine or a histidine-like component than did predentine and osteoid. It is proposed that these differences are involved in initiation of mineralization.
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  • 20
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    Calcified tissue international 16 (1974), S. 169-182 
    ISSN: 1432-0827
    Keywords: Histochemistry ; Alkaline phosphatases ; Calcification ; Bone ; Teeth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Activity of alkaline phosphatases in unfixed cold microtome setions from the lower first molar area of newborn mice was recorded by histochemical methods. A substrate specificity test included the following phosphate compounds: ATP, CTP, GTP, UTP, ADP, AMP, GP, PPi, MDP and naphthol AS-TR phosphate. Intense staining was obtained in osteoblasts, stratum intermedium of the enamel organ and odontoblasts with all the substrates, except PPi and MDP. Staining of skeletal muscle fibres was obtained only with triphosphates as substrates. Addition of-SH groups decreased the hydrolysis of triphosphate compounds in cells involved in mineralization while the hydrolysis of monophosphate was inhibited. In contrast triphosphatase activity in striated muscle was enhanced when-SH compounds were added. Demineralization with EDTA diminished the cytoplasmic staining but induced a nuclear staining in hard tissue forming cells when triphosphates were used as substrates. No cytoplasmic and only slight nuclear staining was seen with GP or AMP as substrates. The triphosphate hydrolyzing capacity of tongue muscle fibres was, however, increased after the decalcification treatment. Addition of Mg2+ ions to the incubation media distinctly lowered the hydrolysis of triphosphates in the investigated tissues whereas the hydrolysis of ADP, AMP, GP and naphthol AS-TR phosphate remained unchanged. In view of the findings the triphosphatase activities at alkaline pH of muscle fibres and of cells related to hard tissue formation are considered to be due to activity of separate enzymes. The orthophosphate liberating enzyme activities at alkaline pH in osteoblasts, stratum intermedium and odontoblasts may be expressions of the catalytic functions of one common enzyme. Furthermore, the results indicate that CaATP might be the substrate used by the alkaline ATPase in mineralizing areas.
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  • 21
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    Calcified tissue international 16 (1974), S. 239-250 
    ISSN: 1432-0827
    Keywords: EHDP ; Bone ; Collagen ; Histology ; Bone chemical analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The purpose of this study was to determine the short-term effects of various systemic doses of disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) on bone organic matrix and to relate these effects to the corresponding dose-related changes in bone mineral. EHDP was administered daily by subcutaneous injection at doses of 0.25, 2.5 and 40 mg/kg body weight for periods of one and two weeks. At both time intervals, rat tibiae were quantitatively analyzed for mineral content (ash, calcium and phosphorus) and for organic matrix content (matrix weight, nitrogen and certain amino acids). The latter data were correlated with semiquantitative histological analyses of the tibiae. Results of this study demonstrate that the short term effects of EHDP on bone chemistry and histology are variable and depend on the systemic dose and the duration of treatment. Systemic doses of 0.25 and 2.5 mg/kg EHDP following daily administration for one week resulted in transitory decreases in bone mineral content compared to controls. Following two weeks of treatment, both of these dose levels resulted in increased bone mineral content and, in addition, the 2.5 mg/kg dose resulted in tibiae which contained more organic matrix compared to control bones. In contrast to the low dose effects, a high systemic dose of EHDP—e.g. 40 mg/kg administered daily for 1 or 2 weeks—appears to act solely by inhibiting mineralization of newly-formed matrix.
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  • 22
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    Calcified tissue international 14 (1974), S. 3-14 
    ISSN: 1432-0827
    Keywords: Hydroxyapatite ; Mineral ; Phase ; Chemistry ; Synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des diagrammes de phase d'équilibre ont été déterminés pour le système CaO-P2O5-H2O en utilisant des techniques de synthèse hydrothermique au cours de variatio nsde température allant de 300–600° et 2 Kb H2O de pression. De l'hydroxyapatite bien cristallisé a été synthétisé et caractérisé. De faibles variations de paramètres de la maille cristalline, liées à la température de synthèse et composition globale du matériel initial, ont été déterminées. Des conditions chimiques précises sont nécessaires pour obtenir de l'apatite, en tant que seule phase solide en équilibre dans la solution. Les résultats de diagramme de phase d'équilibre sont comparés avec ceux obtenus dans des milieux synthétiques.
    Abstract: Zusammenfassung Es wurden Gleichgewichts-Phasendiagramme für das System CaO-P2O5-H2O bestimmt, indem hydrothermale Synthese-Techniken im Temperaturbereich von 300–600° und bei einem Druck von 2 Kb H2O verwendet wurden. Es wurde gut-kristallisiertes Hydroxyapatit erzeugt und charakterisiert. Es wurden geringe Unterschiede in den Parametern der Zelleinheiten festgestellt, welche von der angewandten Temperatur und der Zusammensetzung des Startmaterials abhingen. Es waren genaue chemische Bedingungen nötig, um Apatit als die einzige feste Phase im Gleichgewicht mit der Lösung zu erhalten. Die Resultate der Gleichgewichts-Phasendiagramme werden mit früheren Untersuchungen mit der Synthesetechnik verglichen.
    Notes: Abstract Equilibrium phase diagrams have been determined for the system CaO-P2O5-H2 using hydrothermal synthesis techniques in the temperature range 300–600° and 2 Kb H2O pressure. Well-crystallized hydroxyapatite has been produced and characterized. Small variations in unit cell parameters dependent on temperature of synthesis and bulk composition of the starting materials have been determined. Precise chemical conditions were required to obtain apatite as the only solid phase in equilibrium with solution. Equilibrium phase diagram results are compared with previous synthetic investigations.
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  • 23
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    Calcified tissue international 14 (1974), S. 31-48 
    ISSN: 1432-0827
    Keywords: Bone ; Cartilage ; Calcification ; Collagen ; Phosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une méthode pour l'étude de la croissance des os longs de foetus de rat, en culture d'organe, dans un milieu chimiquement défini, a été mise au point. Les extrémités cartilagineuses et les parties centrales de l'os sont analysées séparément pour leur croissance et minéralisation en étudiant leur contenu en collagène, calcium et phosphate, poids sec, et incorporation de proline marquée en hydroxyproline. La croissance et la minéralisation des parties centrales osseuses sont plus lentes dans un milieu chimiquement défini qu'in vivo. La croissance peut être accélérée en ajoutant au milieu des acides aminés non essentiels, de l'albumine ou du sérum. Les extrémités cartilagineuses présentent une augmentation plus importante en poids et contenu en collagène que les parties centrales et l'adjonction de diverses substances a moins d'effet sur la croissance. La croissance et la minéralisation des parties centrales sont augmentées en élevant la concentration du milieu en phosphate de 1.5 à 4.5 mM, avec ou sans adjonction de sérum ou d'albumine. A une concentration faible de calcium (0.5 mM), la croissance et la minéralisation des parties centrales sont arrêtées. A une concentration faible en magnésium (0.5 mM), la minéralisation est augmentée, mais la croissance est arrêtée.
    Abstract: Zusammenfassung Es wird eine Methode beschrieben, mit welcher das Wachstum der Röhrenknochen von Rattenembryos in einem chemisch bestimmten Medium in Organkultur untersucht werden kann. Die Knorpelenden und Knochenschäfte wurden gesondert auf Wachstum und Mineralisation geprüft, indem Collagen-, Calcium- und Phosphatgehalt, das Trockengewicht und der Einbau von markiertem Prolin in Hydroxyprolin gemessen wurden. Wachstum und Mineralisation des Knochenschaftes waren langsamer in einem chemisch bestimmten Medium als in vivo. Das Wachstum konnte beschleunigt werden, indem dem Medium nicht-essentielle Aminosäuren, Albumin oder Serum beigegeben wurden. Die Knorpelenden zeigten eine viel stärkere Zunahme an Gewicht und Collagengehalt als die Schäfte, und Anreicherung des Mediums hatte weniger Wirkung auf ihr Wachstum. Das Wachstum und die Mineralisation der Knochenschäfte nahmen zu, wenn die Phosphatkonzentration im Medium zwischen 1,5 und 4,5 mM erhöht wurde, und zwar unabhängig davon, ob dem Medium Serum oder Albumin beigegeben wurde oder nicht. Bei niederer Calciumkonzentration (0,5 mM) im Medium wurden Wachstum und Mineralisation der Knochenschäfte beeinträchtigt. Bei niedriger Magnesiumkonzentration (0,5 mM) wurden die Mineralisation erhöht, das Wachstum hingegen gehemmt.
    Notes: Abstract A method for studying the growth of fetal rat long bones in a chemically defined medium in organ culture is described. Cartilage ends and bone shafts were analyzed separately for growth and mineralization by measuring the collagen, calcium, and phosphate content, dry weight, and incorporation of labeled proline into hydroxyproline. Growth and mineralization of the bone shaft were slower in a chemically defined medium thanin vivo. Growth could be enhanced by supplementation of the medium with non-essential amino acids, albumin or serum. Cartilage ends showed a greater increase in weight and collagen content than the shafts, and medium supplements had less effect on their growth. Bone shaft growth and mineralization were enhanced by increasing medium phosphate concentration over a range of 1.5 to 4.5 mM whether or not the medium was supplemented with serum or albumin. At a low medium calcium concentration (0.5 mM) bone shaft growth and mineralization were impaired. At a low magnesium concentration (0.5 mM) mineralization was enhanced, but growth was impaired.
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  • 24
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    Calcified tissue international 14 (1974), S. 83-86 
    ISSN: 1432-0827
    Keywords: Bone ; Reproductive cycle ; Pigeons ; Amorphous ; Crystalline
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une étude de diffraction aux rayons X, au cours du cycle de ponte d'oeufs de pigeon, a été entreprise pour évaluer les modifications de la proportion entre les phases de phosphate de calcium amorphe et apatitique. La partie médullaire du fémur de pigeon est totalement résorbée au cours du cycle de reproduction. Au cours de ce processus, l'os contenant plus de phosphate de calcium amorphe est résorbé avant celui qui est riche en cristaux. Ainsi l'os à contenu amorphe plus élevé est métaboliquement plus actif que l'os riche en cristaux. L'os médullaire qui peut être induit chez le pigeon mâle par traitement aux oestrogènes, ressemble, au point de vue diffraction aux rayons X, à l'os médullaire des femelles au début du cycle de reproduction.
    Abstract: Zusammenfassung Mittels Röntgen-Diffraktion wurden die Veränderungen in den prozentualen Mengen der amorphen und der kristallinen Calciumphosphat-Phasen im Femur der Taube während des Eierlegens untersucht. Der medulläre Teil des Taubenfemur wird während des Fortpflanzungscyclus vollständig resorbiert. Dabei wird der Teil des Knochens, der mehr amorphes Calciumphosphat enthält, vor dem kristallreichen Teil resorbiert. Daraus folgt, daß der Stoffwechsel des Knochens mit höherem amorphem Gehalt aktiver ist als derjenige des Knochens mit höherem kristallinem Gehalt. Weiter konnte mittels Röntgendiffraktion festgestellt werden, daß der medulläre Knochen, welcher in männlichen Tauben durch Oestrogenbehandlung erzeugt werden kann, dem weiblichen medullären Knochen am Anfang des Fortpflanzungscyclus gleicht.
    Notes: Abstract This is an X-ray diffraction study of the changes in the proportional amounts of the amorphous and apatitic calcium phosphate phases in pigeon femur during the egg-laying cycle. The medullary portion of pigeon femur is completely resorbed in the reproductive cycle. In this process the portion of bone which is higher in amorphous calcium phosphate is resorbed before the crystal-rich portion. Thus, bone with a higher amorphous content is more metabolically active than more crystalline bone. Finally, medullary bone which can be produced in male pigeons by estrogen treatment was found by X-ray diffraction to resemble female medullary bone at the beginning of the reproductive cycle.
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  • 25
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    Calcified tissue international 14 (1974), S. 49-58 
    ISSN: 1432-0827
    Keywords: Glycosaminoglycans ; Intracellular ; Chondrocytes ; Cartilage ; Embryonal ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les glycosaminoglycanes acides de chondrocytes isolés, en suspension, sont étudiés. La matrice extracellulaire est isolée par traitement successif à la trypsine et à la collagénase d'os d'embryons de poulet, âges de 15 à 17 jours. Après digestion à la papaïne et élimination des acides nucléïques par traitement à la DNAse et à la RNAse, les glycosaminoglycanes sont précipités par le CPC et isolés sous la forme de leur sel sodique. Les analyses des propriétés de solubilité du CP glycosaminoglycane par le microfractionnement d'Antonopoulos et coll. (1964) révèlent la présence de glycosaminoglycanes intracellulaires de poids moléculaire et/ou de densité de charge nettement plus faibles que ceux de la matrice extracellulaire. Sur des électrophorèses de microzones, une petite partie des glycosaminoglycanes intracellulaires isolées présente une mobilité similaire à celle des solutions témoins de sulfate de chondroitine, alors que la portion la plus importante ne se déplace qu'à mi-distance de l'acide hyaluronique et montre une large trainée, indiquant une faible charge négative. Ce fait semble dû à une perte de l'influence de régulation des composants matriciels éliminées, sur la synthèse des glycosaminoglycanes et au rejet dans le milieu de glycosaminoglycanes initialement synthétisé. Pour la première fois, une faible quantité d'acide hyaluronique intracellulaire est mise en évidence par une mobilité électrophorétique typique.
    Abstract: Zusammenfassung Saure Glykosaminoglycane von isolierten Chondrocyten in Suspension wurden untersucht, und die extracelluläre Matrix von 15–17 Tage alten Knochen von Kükenembryos wurde durch die aneinanderfolgende Behandlung mit Trypsin und Collagenase erhalten. Nach der Papain-Verdauung und Beseitigung der Nucleinsäuren mittels DNS- und RNSase-Behandlung wurden die Glykosaminoglycane mit CPC gefällt und als Natriumsalz isoliert. Untersuchungen der Löslichkeit der CP-Glykosaminoglycane mittels des Mikrofraktionierungsverfahrens von Antonopouloset al. (1964) zeigten intracelluläre Glykosaminoglycane mit meist niedrigerem Molekulargewicht und/oder niedrigerer Ladungsdichte als diejenigen der extracellulären Matrix. In der Mikro-Zonen-Elektrophorese zeigte nur ein kleiner Teil der isolierten intracellulären Glykosaminoglycane eine Mobilität, die dem Standard des Chondroitinsulfats entsprach. Der größte Teil legte nur die halbe Distanz der Hyaluronsäure zurück und zeigte breites “Tailing”, was auf eine niedrigere negative Ladung hindeutet. Die Ursache dafür wurde interpretiert als Verlust eines regulierenden Einflusses der entfernten Matrixkomponenten auf die Glykosaminoglycan-Synthese und als Abgabe ursprünglich synthetisierter Glykosaminoglycane in das Medium. Es wurde erstmals, anhand der typischen elektrophoretischen Beweglichkeit, eine kleine Menge von intracellulärer Hyaluronsäure nachgewiesen.
    Notes: Abstract Acid glycosaminoglycans were studied from isolated chondrocytes in suspension and the extracellular matrix obtained by sequential trypsin and collagenase treatment of 15- to 17-day-old embryonic chick bone. After papain digestion and removal of the nucleic acids by treatment with DNAse and RNAse, the glycosaminoglycans were precipitated by CPC and isolated as their sodium salt. Analyses of the CP-glycosaminoglycan solubility properties with the microfractionation procedure of Antonopouloset al. (1964) showed intracellular glycosaminoglycans of predominantly lower molecular weight and/or charge density than those of extracellular matrix. On micro-zone electrophoresis, only a minor part of the isolated intracellular glycosaminoglycans showed a mobility similar to that of the chondroitin sulphate standards while the major part moved only half the distance of hyaluronic acid and showed broad tailing, indicating a low negative charge. This was considered to be due to loss of a regulatory influence of the removed matrix components upon glycosaminoglycan synthesis and to release of originally synthesized glycosaminoglycans into the medium. For the first time, a small amount of intracellular hyaluronic acid was shown by typical electrophoretic mobility.
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  • 26
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    Calcified tissue international 14 (1974), S. 161-168 
    ISSN: 1432-0827
    Keywords: Diphosphonates ; Osteoid Synthesis ; Hydroxyproline ; Bone ; Formation-Resorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Deux diphosphonates, à savoir l'éthane-1-hydroxy-1,1-diphosphonate (EHDP) et le dichlorométhane diphosphonate (Cl2MDP), inhibentin vitro la résorption osseuse qui est stimulée soit par la parathormone ou par voie intrinsèque dans un système de culture d'os en remaniement. Alors que le Cl2MDP est plus efficace que l'EHDP dans l'inhibition de la résorption, ce fait semble en rapport avec une diminution de le formation de tissu ostéoide. Il s'en suit une diminution biochimique et morphologique marquée du remaniement osseux avec le Cl2MDP à une concentration équivalent à 10 μg de phosphore/ml de milieu de culture. La différence d'activité entre l'EHDP et le Cl2MDP peu être liée à leur affinité relative pour les surfaces du minéral osseux et, par suite, à leur concentration effective dans la fluide extracellulaire osseux. Les diphosphonates pourraient aussi affecter l'os indirectement si l'on admet que le degré de minéralisation de la matrice est important pour l'induction et la régulation de l'activité ostéoblastique dans l'os en voie de remaniement.
    Abstract: Zusammenfassung Zwei Diphosphonate, Aethan-1-hydroxy-1,1-diphosphonat (EHDP) und Dichloromethylen-Diphosphonat (Cl2MDP) hemmen in vitro die Knochenresorption. Diese wird verfolgt entweder mittels Stimulierung durch PH oder in einem Kultursystem, in welchem normale Knochenneubildung stattfindet. Während Cl2MDP die Resorption wirksamer hemmt als EHDP, scheint es ebenfalls eine Verminderung der Osteoidbildung zu verursachen. Dies bewirkt eine deutliche biochemische und morphologische Herabsetzung der Knochenneubildung, bei einer Cl2MDP-Konzentration von 10 μg Phosphor/ml Kulturmedium. Die unterschiedliche Wirksamkeit von EHDP und Cl2MDP läßt sich wahrscheinlich auf ihre verschiedenen Affinitäten zu der Oberfläche des Knochenminerals und somit auf ihre tatsächliche freie Konzentration in der extracellulären Flüssigkeit des Knochens zurückführen. Ausgehend von der Annahme, daß die Anregung und Regulierung der Osteoblasten-Aktivität bei der Knochenneubildung vom Ausmaß der Matrix-Mineralisation bestimmt wird, läßt sich die Vermutung aufstellen, daß die Diphosphonate die Knochenbildung auch indirekt beeinflussen können.
    Notes: Abstract Two Diphosphonates, ethane-1-hydroxy-1,1-diphosphonate (EHDP) and dichloromethylene diphosphonate (Cl2MDP) inhibitin vitro bone resorption, which is either stimulated by parathyroid hormone or intrinsic in a bone remodeling culture system. While Cl2MDP is more effective than EHDP in inhibiting resorption, it also appears to result in a related diminution in osteoid formation. This effect causes a marked biochemical and morphological depression of bone remodelling with Cl2MDP at a concentration equivalent to 10-μg-phosphorus/ml of culture medium. The difference in activity between EHDP and Cl2MDP may be related to their relative affinities for the bone mineral surfaces and hence their effective free concentration in the bone extracellular fluid. It is hypothesized that diphosphonates may also affect bone formation indirectly if one assumes that the degree of mineralization of the matrix is important in the induction and regulation of osteoblastic activity in remodelling bone.
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  • 27
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    Calcified tissue international 14 (1974), S. 195-210 
    ISSN: 1432-0827
    Keywords: Bone ; Fossil bone ; Bone resorption ; Boring channels ; Fungi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'action ostéoclasique exercée par un champignon appartenant au genreMucor sur des os inhumés a été étudiée au point de vue morphologique. L'érosion ossuse donne lieu à des cavités et à des canalicules de percement, dont les parois apparaissent régulièrement calcifiées jusqu'a l'interface avec le champignon, ce qui veut dire que la réabsorption intéresse en même temps la matrice organique et les cristallites. Au contraire des ostéoclastes, la membrane du champignon ne montre jamais un bord en brosse au niveau des points où elle se met en contact avec le tissu osseux. De même l'examen au microscope électronique ne décele apparemment aucun passage de matériel dans l'épaisseur de la membrane ainsi que dans le protoplasme du champignon, ce qui porte à admettre qu'un processus de solubilisation a lieu au préalable. Seulement dans le cas où le champignon présente des phénomènes de vieillissement, des données font prévoir la libération d'une substance capable de décalcifier la matrice organique. Tous les résultats ci-dessu sont discutés de façon analytique et comparés à ceux fournis par les canalicules de percement des os fossiles.
    Abstract: Zusammenfassung Die osteoklastische Aktivität eines der Mucor-Gattung angehörenden Pilzes auf begrabene Knochen wurde morphologisch untersucht. Die Wirkung dieses Pilzes in der Knochenerosion führt zu Resorptionshöhlen und Bohrkanälen, deren Wände scharf und bis zur Oberfläche gut verkalkt sind; das bedeutet, daß die Knochenresorption gleichzeitig die Kristalliten und die organische Matrix in Mitleidenschaft zieht. Im Gegensatz zu den normalen Osteoklasten zeigt die Membran des Pilzes an den Berührungsstellen mit dem Knochengewebe keinen Bürstensaum. Ebenfalls läßt eine elektronenmikroskopische Untersuchung keinen Durchgang von Substanzen durch die Membran und das Protoplasma des Pilzes erkennen, so daß man annehmen muß, daß vorher ein Auflösungsprozeß stattgefunden hat. Die Vermutung liegt nahe, daß eine Substanz, welche den anorganischen Knochenanteil aufzulösen vermag, sich
    Notes: Abstract A morphological investigation has been carried out on the osteoclastic activity revealed by a fungus of theMucor genus on buried bone. The hallmark of its activity in eroding bone is the finding of resorption pits and boring channels whose walls are sharp and well calcified up to the free edge, suggesting that bone resorption affects crystallites and the organic matrix simultaneously. Unlike normal osteoclastic cells, the fungal membrane in contact with the bone shows no brush border. As the electron microscope reveals no migration of material to the fungal membrane and its protoplasm, the view is expressed that the material which reaches the fungus has been previously solubilized. There is good reason for supposing that a substance capable of solubilizing the inorganic bone fraction spreads freely through bone tissue, decalcifying the matrix, where, but only where, hyphae show the effects of ageing. All the findings are thoroughly discussed and compared with those furnished by the boring channels in fossil bone.
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  • 28
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    Calcified tissue international 14 (1974), S. 245-249 
    ISSN: 1432-0827
    Keywords: Bone ; Repair ; Histology ; Implant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Um modèle expérimental, destiné à l'étude de la cicatrisation osseuse, est mis au point. Il permet d'obtenir du tissu osseux pour des études qualitatives et quantitatives. Des tubes de nylon sont implantés dans l'alvéole d'incisives extraites de cobayes. Une étude histologique de coupes longitudinales montrent que les implants de nylon ne retardent pas la cicatrisation des lésions osseuses et contiennent suffisamment de tissus pour des études biochimiques.
    Abstract: Zusammenfassung Es wurde ein Modellsystem zum Studium der Knochenwiederherstellung entwickelt, mit Hilfe dessen man von dem sich wiederbildenden Knochengewebe Proben für die qualitative und quantitative Bestimmung entnehmen kann. Dazu wurden in die Alveolen extrahierter Schneidezähne von Meerschweinchen Nylonschläuchlein eingelegt. Die histologische Untersuchung von Längsschnitten zeigte, daß die Nylonimplantate den Wiederherstellungsprozeß des Knochendefektes nicht verzögerten, daß sie herauspräpariert werden konnten und genügend Gewebe enthielten, um für biochemische Untersuchungen verwendet zu werden.
    Notes: Abstract A model system for the study of bone repair has been developed which allows the recovery of repairing osseous tissue for qualitative and quantitative evaluation. Nylon tubes were implanted in the alveoli of extracted incisors in guinea pigs. A histological study of longitudinal sections showed that the nylon implants did not delay the reparative processes of the bony defect, could be dissected out and contained sufficient tissue to be used in biochemical studies.
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  • 29
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    Calcified tissue international 15 (1974), S. 45-54 
    ISSN: 1432-0827
    Keywords: Bone ; Fluoride ; Microhardness ; Mineralization ; Strength
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Microhardness was measured in sampling sites in the tibial diaphysis of control rats that received less than 1 ppm fluoride in the drinking water, and experimental rats that received 30, 90 and 120 ppm fluoride in the drinking water for 17 days. The latter dose was toxic, as evidenced by a decreased final body weight in this group. By means of tetracycline labelling, it was possible to measure bone hardness in four zones of increasing bone age: I) 3 days, II) 8 days, III) 13 days and IV) 22 days. Zones I through III represented bone formed during fluoride treatment, and Zone IV bone formed before fluoride treatment. In the control group, microhardness increased from Zone I to II, probably because mineral concentration was relatively low in Zone I, and remained constant thereafter. In the 90 and 120 ppm fluoride-treated groups, maximum microhardness was not achieved until Zone III. This delay was probably due to the fact that fluoride in large doses inhibits the rate of mineralization. In the 30 ppm fluoride-treated group, there was no delay in achievement of maximum microhardness; microhardness values in Zones I and III were greater than those in control animals, and microhardness in Zone III was higher than that in Zone IV. These results show that: 1) bone microhardness is increased in bone formed during fluoride treatment in rats given 30 ppm fluoride in the drinking water, 2) toxic doses of fluoride delay, but do not prevent achievement of normal maximum microhardness, and 3) changes in microhardness are seen only in bone formed during fluoride treatment.
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  • 30
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    Calcified tissue international 15 (1974), S. 71-76 
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Deficiency ; Osteoporosis ; Species
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract 21-day-old rats and mice were fed a low-calcium diet (0.02% Ca) or a normal diet (0.5% Ca) for two weeks. Administration of the low-calcium diet resulted in greater decreases in femur ash content and serum calcium in rats than in mice. Microscopic examination revealed that metatarsal bones from a majority of the rats fed a low-calcium diet displayed moderate or severe osteoporotic changes whereas bones from mice were either normal or displayed only slight osteoporotic changes under the same experimental conditions. These results indicate that mice are better able to adapt to a low-calcium diet than are rats of the same age.
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  • 31
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    Calcified tissue international 15 (1974), S. 111-123 
    ISSN: 1432-0827
    Keywords: Fluoride ; Calcium ; Phosphorus ; Bone ; Periosteum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une méthode d'estimation de la surface périostée de l'os en voie de minéralisation est mise au point et appliquée à des porcs recevant 2 mg de fluorure par Kg de poids et par jour pendant 10 mois. L'adjonction de fluor intervient significativement dans le cas d'une déficience en calcium et phosphore. La surface en voie de minéralisation est nettement réduite. Lorsque le calcium et phosphore sont normaux, l'addition de fluorure augmente nettement la surface en voie de minéralisation. Le microscope électronique à balayage permet de mettre en évidence au niveau de l'os une perte de l'orientation longitudinale des fibers et des surfaces poreuses irrégulières chez les animaux soumis au floor.
    Abstract: Zusammenfassung Zur Bestimmung der verkalkenden Oberfläche des Periosts wurde eine Methode entwickelt und bei Schweinen angewandt, welche während etwa 10 Monaten 2 mg Fluorid per kg Körpergewicht und per Tag erhalten hatten. Dieses zusätzliche Fluorid hatte eine signifikante Wirkung bei Calcium- und Phosphormangel. Die verkalkende Oberfläuche ging stark zurück, während jedoch, wenn kein Calcium- und Phosphormangel bestand, das zusätzliche Fluorid die verkalkende Oberfläche vergrößerte. Beobachtungen mit dem Raster-Elektronenmikroskop am Knochen zeigten, daß die Längsorientierung der Fasern fehlte und daß Knochen von mit Fluorid gefütterten Tieren unregelmäßige, poröse Oberflächen aufwiesen.
    Notes: Abstract A method for estimating periosteal bone-mineralizing surface was developed and applied to swine fed 2 mg of fluoride/kg of body weight/day for about 10 months. Added fluoride interacted significantly with calcium and phosphorus deficiency. Mineralizing surface was greatly reduced, whereas when calcium and phosphorus were adequate, added fluoride greatly increased mineralizing surface. Scanning electron microscope observations of bone disclosed loss of longitudinal orientation of fibers and irregular porous surfaces in bone from fluoride-fed animals.
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  • 32
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    Calcified tissue international 15 (1974), S. 125-132 
    ISSN: 1432-0827
    Keywords: Bone ; Collagen ; Formic Acid ; Crosslinkages ; Cyanogen Bromide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Bovine and chicken bone collagens have been solubilized and presumably denatured (gelatin) by treatment of demineralized, powdered tissue with 70% formic acid. Short periods of extraction such as four hours at 30°, conditions commonly used during cyanogen bromide cleavage of collagen, solubilized 50% and 15% of the chicken and bovine bone collagens respectively. Treatment of the tissues with sodium borohydride partially inhibited the extraction of collagen from chicken bone, but had little effect on the extraction of calf bone collagen. The heterogeneity of the bone gelatin from both species on disc electrophoretic analysis suggested that peptide bonds had been cleaved in some of the collagen chains during exposure to formic acid, thus facilitating the solubilization of the bone collagen as the gelatin. Analysis of the collagen extracted from chicken bone for reducible crosslinks indicated that a large proportion of these bonds had remained intact, in contrast to a previous finding that most of these crosslinks were destroyed in bone gelatin extracted by 4 M CaCl2, at pH 7.0. The stability of the major reducible crosslinks in bone collagen to severe acid conditions may explain in part some of its unique properties, such as its failure to swell or be solubilized in dilute acid, which distinguish it from soft tissue collagens.
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  • 33
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    Calcified tissue international 15 (1974), S. 173-179 
    ISSN: 1432-0827
    Keywords: Bone ; Strength ; Tension ; Compression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract A simple method is described for visualising regions of bone that have yielded in tension. The appearance of such regions is quite different from that of regions that have yielded in compression. The significance of these findings is discussed.
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  • 34
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    Calcified tissue international 15 (1974), S. 189-199 
    ISSN: 1432-0827
    Keywords: Bone ; Densitometry ; X-ray ; Radiation ; Specific gravity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract X-ray and γ-radiation techniques for evaluating bone density under field conditions in cattle and sheep were investigated. The best results were obtained from measurements of the density of radiographs of tibial tarsal bones of sheep and from the absorption of γ-radiation of the tibial tarsal bones of sheep and the fibular tarsal bones of cattle. The γ-radiation techniques may be carried out rapidly under field conditions (up to 30 animals an hour) and bone density can be estimated with a standard error of 0.042–0.046 SG units.
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  • 35
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    Calcified tissue international 15 (1974), S. 181-188 
    ISSN: 1432-0827
    Keywords: Biopsy ; Bone ; Calcium ; Phosphorus ; Specific gravity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The specific gravities (SG) of bone samples taken from various parts of the skeleton of cattle and sheep were determined gravimetrically in an effort to establish which sites give the most reproducible and uniform results, so that between animals, or sequential within animals, comparisons may be made with maximum sensitivity. Samples obtained from the mandible of sheep and the rib of cattle and sheep were found to be too variable to be useful for most purposes. Best results were obtained using whole bones which are easily prepared, such as the tibial tarsal bones of cattle and sheep and the fibular tarsal bone of cattle. These bones gave within animal deviations of 0.012–0.024 SG units, when comparing left bone with right bone. Slightly higher values were obtained for coccygeal vertebrae from cattle and distal metacarpal condyles from sheep. SG was related to ash, calcium and phosphorus contents of coccygeal vertebrae of cattle and it is considered that, for survey work on the mineral status of the bones of cattle and sheep, the correlations are sufficiently high to make their determination unnecessary. This will enable radiation determined SG techniques to be applied to this type of work.
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  • 36
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    Calcified tissue international 16 (1974), S. 335-338 
    ISSN: 1432-0827
    Keywords: Fluoride ; Plasma ; Bone ; Age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Fluoride analyses were carried out on blood samples from fasting subjects and on iliac crest biopsies obtained at autopsy, using an ion-selective electrode. Correlation coefficients of 0.53 and 0.67 respectively were determined between plasma fluoride and age, and bone fluoride and age. The proportionate rates of increase of fluoride levels in plasma and bone with age and the similarity of the correlation coefficients suggest that the fasting plasma fluoride concentration is in equilibrium with bone fluoride.
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  • 37
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    Calcified tissue international 16 (1974), S. 305-313 
    ISSN: 1432-0827
    Keywords: Osteosarcoma ; Lyophilization ; Induction ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Bone formation was induced by iso- and home-implantation of lyophilized mouse osteosarcoma, in which an osteogenic activity is observed. The inducing factor is present exclusively in osteosarcoma, such as BF and Gardner, while not in non-osteogenic tumors, such as Ehrlich ascites tumor (2N & 4N) or mouse mammary carcinoma. The production of bone was rapid and extensive. It usually occurred within 21 days, and the time required for osteogenesis was almost the same as that reported in previous studies.
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  • 38
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    Cell & tissue research 148 (1974), S. 11-26 
    ISSN: 1432-0878
    Keywords: Dentine ; Bone ; Collagen structure ; Collagen mineralization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary It has been previously shown that the distances between the nuclei within the collagen bundles of mineralizing tissues were in good agreement with the repeat distances of the cross-banding pattern of collagen, which supports the assumption that the distances between the mineral deposits reflect to a good approximation the distances between nucleation centres on the collagen macromolecule. However, the lateral separation of the nuclei were significantly higher than the distances between close-packed triple helices. Recently a new model of collagen aggregation has been proposed in which the smallest morphological units are subfibrils (Ø approx. 39 Å) packed in tetragonal array. This led us to measure once again the lateral separation between a) close-packed calcium phosphate needles lying in bundles at (1) the mineralizing front of mantle dentine and (2) at the mineralizing front of rat tail bone, and b) between the uranyl-lead nuclei produced in the staining of rat tail tendon. The mean lateral distances separating these nuclei fell within the range of 39–47 Å, which is a little higher than the distances of 39 Å which separate the microholes between the subfibrils in the tetragonal packing model, which are regarded as the likely sites of nucleation. If, however, it is assumed that the forces generated during mineralization can cause the collagen fibres to swell, then the lateral separation of the nuclei and the distances between the microholes would correspond very closely.
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  • 39
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    Langenbeck's archives of surgery 335 (1974), S. 263-272 
    ISSN: 1435-2451
    Keywords: Coating ; Blood-Compatibility ; Gels ; Chemistry ; Hydrophilic Monomers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Es wurden die wichtigsten Arbeiten über hydrophile Gele ausgewählt und im Hinblick auf ihre Eignung als aussichtsreiche Blut-Kontaktmaterialien betrachtet. Die Reaktionsbedingungen werden beschrieben, und die gegenseitige Wechselwirkung von Blut und Kunststoff wird dargestellt.
    Notes: Summary By the selection of the relevant sources it is attempted to present the contemporary research situation of the hydrophilic polymeric gels as promising materials for the contact with the blood in the organism. The reaction conditions of the current syntheses are described as well as the mutual interactions of the blood and the contact surface.
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  • 40
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    Biopolymers 13 (1974), S. 1-27 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: We describe a method for determining chemical kinetic constants and diffusion coefficients by measuring the rates of decay of spontaneous concentration fluctuations. The equilibrium of the system is not disturbed during the measurement. We measure the number of molecules of a specified type in a defined open volume as a function of time and compute the time course of the deviations from the thermodynamic mean concentration. The method is based on the principle that the rates of decay of spontaneous microscopic fluctuations are determined by the same phenomenological rate coefficients as those of macroscopic departures from equilibrium which result from external perturbations. Hence, an analysis of fluctuations yields the same chemical rate constants and diffusion coefficients as are measured by conventional procedures. In practice the number of the specified molecules is measured by a property such as absorbance or fluorescence which is specific and sensitive to chemical change. The sample volume is defined by a light beam which traverses the cell. As the molecules appear in or disappear from the light beam, either due to diffusion or chemical reaction, their concentration fluctuations give rise to corresponding fluctuations of the intensity of absorbed or emitted light. This paper presents the theory needed to derive chemical rate constants and diffusion coefficients from these fluctuations in light intensity. The theory is applied to three examples of general interest: pure diffusion in the absence of chemical reaction; the binding of a small rapidly diffusing ligand to a larger slowly diffusing macromolecule; and a unimolecular isomerization. The method should be especially useful in studying highly cooperative systems, relatively noncooperative systems with intermediate states closely spaced in free energy, small systems, and systems not readily subject to perturbations of state.
    Additional Material: 3 Ill.
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  • 41
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    Biopolymers 13 (1974), S. 97-114 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The molecular conformations and dynamics of poly(L-prolyl), poly(hydroxyl-L-prolyl), poly(L-prolyl-glycyl), poly(hydroxyl-L-prolyl), and poly(glycyl-glycyl-L-prolyl-glycyl), in aqueous solution, have been studied using 13C pulse Fourier transform nmr spectroscopy. From a measurement of the intensities of major and minor resonances in the spectra of the copolypeptides, it was determined that 15-20% of the glycyl-prolyl and glycyl-hydroxyprolyl peptide bonds are cis. Effective rotational correlation times (τeff), obtained from measurements of spin-lattice relaxation times (T1) of individual backbone and side-chain carbons, demonstrated that backbone reorientation is approximately isotropic for the five polypeptides and is characterized by correlation times of ca. 0.3-0.6 nanoseconds as a result of rapid segmental motion. In a given polypeptide glycyl and pyrrolidine residues were found to have the same backbone correlation times, but backbone carbon τeff values did decrease as the glycyl content of the peptides increased. A semi-quantitative analysis of Cβ, Cγ, and Cδ correlation times suggests that rapid ring motion in both prolyl and hydroxyprolyl involves primarily Cγ and Cβ, with the prolyl ring being more mobile than the hydroxyprolyl ring.
    Additional Material: 3 Ill.
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  • 42
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    Biopolymers 13 (1974), S. 237-238 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: No. Abstract.
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  • 43
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    Biopolymers 13 (1974), S. 217-226 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A model for the flexibility of DNA is proposed that is based on discrete variations in the direction of propagation in going from one subunit to the next. Expansion of the local free energy in terms of the local bending gives a Gaussian distribution function. The assumption of the independence of local bends on neighbors lead to very simple formulae for the persistence length and the characteristic ratio. Emphasis, however, is placed on the application of the formulae for molecules of finite size where the persistence length and C∞ are not defined. The formulae are worked out for two models, which should serve as limits for the real physical situation.
    Additional Material: 2 Ill.
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  • 44
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    Biopolymers 13 (1974), S. 735-743 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: If dispersion effects are not taken into account, the dimensions of particles in suspension estimated from spectrophotometric data (slope of the log OD versus log λ plot) are too low. By measuring dispersion curves for the medium and for the suspension, a consistent estimate of the size of latex particles of different diameter is obtained from the OD data.
    Additional Material: 3 Ill.
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  • 45
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    Biopolymers 13 (1974), S. 765-768 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The helix-coil transition of poly-N5-(3-hydroxypropyl)-L-glutamine in methanol-water solutions has been observed using 13C nuclear magnetic resonance. Two signals appear in the α-CH region in the course of the transition; this is in contrast with previous proton magnetic resonance experiments carried out on this polymer in the same solvent system.
    Additional Material: 1 Ill.
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  • 46
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    Biopolymers 13 (1974), S. 779-790 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: X-ray diffraction and stereochemical analyses were used to study the hydrated structure of the helical amylose polymorph having a fiber repeat spacing of 8 Å. Intensity calculations using computer models confirmed six as the number of residues per turn and supported the space group P212121. Both diffraction intensity and stereochemical methods indicate the suitability of residue G3 from the potassium acetate complex of cyclohexaamylose as opposed to residues with longer O(4)-O(1) vectors. Comparison of the present results with those obtained for V amylose dehydrate indicated no major conformational differences between the two helix skeletons. A net helical rotation of approximately 30° accompanied the hydrate-dehydrate transition and the rotational position in the hydrate allowed packing that was less close. Hydration water molecules were not located; noncarbohydrate peaks on the electron density maps were primarily due to Fourier series termination errors.
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  • 47
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    Biopolymers 13 (1974), S. 843-852 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: We have measured the circular dichroism spectra and have obtained melting curves for samples of purified satellite DNA's from three species of crab, Cancer borealis, Gecarcinus lateralis, and Callinectes sapidus. Although all three satellite DNA's have the same density in neutral CsCl, their circular dichroism spectra and melting and reassociation characteristics are different, as are other properties of these DNA's. The circular dichroism spectra are analyzed to give estimates of nearest-neighbor frequencies. We estimate that about 90% of the Cancer borealis satellite DNA and over 80% of the Gecarcinus lateralis satellite DNA consist of d(AT)n:d(AT)n sequences, while the serially repeating unit of the Callinectes sapidus satellite DNA may not contain any extended d(AT)n:d(AT)n sequences. The results from the circular dichroism spectra for the Cancer borealis, Gecarcinus lateralis, and the Callinectes sapidus satellite DNA's are in agreement with base composition analyses, which give 2.4, 8, and 25% G+C, respectively, for these DNA's.
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  • 48
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    Biopolymers 13 (1974), S. 879-890 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Samples of epichlorohydrin crosslinked potato starch were prepared by using a high ratio of starch to water and alkali concentration below the gelatinization level. This yielded crosslinked samples that were partially crystalline, and where the number of crosslinks could be varied between 1 and 20 crosslinks per 100 anhydroglucose units. The degree of swelling varied regularly with degree of crosslinking, and the molecular weight between crosslinks Mc as derived from swelling data in a good swelling agent compared favorably with Mc derived from chemical analysis.From the heat of gelatinization of the crosslinked starches, as observed in a differential scanning calorimeter, for gelatinization in glycerol, water, and dimethylsulfoxide, a model for the gel state of the crosslinked starch is proposed. It is concluded that the entropy of melting is the determining factor in establishing the temperature of gelatinization.
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  • 49
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    Biopolymers 13 (1974), S. 909-917 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Circular dichroism studies of the interaction of β-cyclodextrin with a series of eight dinucleoside phosphates, and with 3′-AMP are reported. From results with ApU and ApA, it is shown that β-cyclodextrin binding is sensitive to dinucleoside stacking in approximately the same way as optical measures of stacking. Some qualitative uses of β-cyclodextrin binding are suggested, based on the fact that the change in the CD spectrum caused by cyclodextrin binding is unique to each of the dinucleosides studied. Hoffman and Bock have previously suggested the use of β-cyclodextrin as a probe of nucleic acid structure. Their work indicated that only binding to adenosine and inosine would have to be considered. The present paper shows that binding to other bases cannot be neglected, and will impose serious restrictions on the use of β-cyclodextrin as a structural probe.
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  • 50
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    Biopolymers 13 (1974), S. 987-993 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Poly(1,N6-etheno-2-aza-adenylic acid) [poly(2-aza-εA)] was synthesized from 1,N6-etheno-2-aza-adenosine 5′-diphosphate (2-aza-εADP) and Escherichia coli polynucleotide phosphorylase. The values Km = 1.02 mM, V = 1.06 μmol hr-1 enzyme unit-1 were found for the polymerization reaction. In contrast to polyadenylic acid, this novel fluorescent polymer has a random structure in solution. The application of the 2-aza-εADP for localization of polynucleotide phosphorylase was also described.
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  • 51
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    Biopolymers 13 (1974), S. 1011-1022 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A series of L-methionine oligomers [BOC-(Met)n-OMe] (n = 2-7) and the corresponding diastereomeric di- and tripeptides were synthesized using the mixed anhydride method. Oligomers prepared in this manner were optically pure and were obtained in reasonable yield. Preliminary optical examination of the peptides suggests that secondary structures may begin forming in the pentamer or hexamer in trifluoroethanol. BOC-(Met)4-OMe and BOC-(Met)5-OMe were also synthesized using an insoluble resin containing BOC-L-methionine as the nitrophenol active ester.
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  • 52
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Conformational studies of a series of oligopeptides (from the tripeptide to the octadecapeptide) with the repeating sequence L-norvalyl-glycyl-L-proline and a polytripeptide with this sequence are reported. By means of chiroptical techniques, unordered conformations are found for all oligopeptides in water, trifluoroethanol, and ethylene glycol and for the water-insoluble polymer in trifluoroethanol. In ethylene glycol the polymer assumes a collagen-like structure. Infrared studies indicate that all the oligomers, in contrast to the polymer, are unordered in the solid state.
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  • 53
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    Biopolymers 13 (1974), S. 1109-1116 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Uncharged poly(Nε-methyl-L-lysine) (PMLL) and its isomer, poly(Nδ-ethyl-L-ornithine) (PELO), in alkaline solution (pH ca. 12) undergo a helix-to-β transition upon mild heating at 50°C or higher in a manner similar to that of poly(L-lysine) (PLL). The rate of conversion follows the order: PMLL 〈 PELO 〈 PLL. The helix can be regenerated upon cooling near zero degrees, for instance, after more than 12 hr at 2°C. At concentrations less than 0.02% the β form is intramolecular, but at higher concentrations both intra- and intermolecular β forms are generated. Poly(Nδ-methyl-L-ornithine) (PMLO), an isomer of PLL, behaves like poly(L-ornithine); uncharged PMLO in alkaline solution is partially helical and becomes disordered at elevated temperatures.
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  • 54
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    Biopolymers 13 (1974), S. 1161-1171 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: In order to study the effect of side-chain length on the conformation of polypeptides, conformational changes of various ionic polypeptides with various lengths of side chain, poly-Nε-glutaryl-L-lysine (PGL), poly-Nδ-glutaryl-L-ornithine (PGO), poly-Nε-succinyl-L-lysine (PSL), and poly-Nδ-succinyl-L-ornithine (PGO), were investigated by ORD, potentiometric titration, and dilatometric measurements in aqueous solution. The results of optical rotation and potentiometric titration measurements indicate strongly that the α-helix stability increases in the sequence PSO 〈 PSL ∼ PGO 〈 PGL, which corresponds to increased side-chain length. The volume change associated with the helix-coil transition also increased in the above sequence. This series of polymers seems to be more hydrophobic compared with poly-L-glutamic acid or poly-L-lysine, as suggested from the values of enthalpy and entropy changes for coil-helix transitions.
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  • 55
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    Biopolymers 13 (1974), S. 1195-1203 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The conformational changes of poly-Nε-glutaryl-L-lysine (PGL) and poly-Nε-succinyl-L-lysine (PSL) in various salt solutions were studied by use of ORD and potentiometric titration measurements. The addition of alkali metal salts to the fully ionized PGL or PSL solution caused helix formation. The helical content of the polymers increases in the following sequences: at salt concentration 0-2 M, CsCl 〈 KCl 〈 LiCl 〈 NaCl; and at 2-3 M, LiCl 〈 CsCl 〈 KCl ∼ NaCl. The preferential binding of the solvent components with various alkali metal salts of PGL or PSL was measured in LiCl, NaCl, and KCl solutions by means of equilibrium dialysis and differential refractometry. It was found that with increasing salt concentration, the polymers were preferentially hydrated in NaCl and KCl soultions; however the salt was preferentially bound to the polymers in LiCl solution. Such preferential binding was suggested to be closely related to conformational change. The addition of CaCl2 to polymer solutions led to the stabilization of the helical structure of PGL or PSL.
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  • 56
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    Biopolymers 13 (1974), S. 1261-1267 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Refractive indices of bovine serum albumin solutions in saline and in 2-chloro ethanol are measured for the spectral region 1770-10000 Å by a reflectance method. A procedure for the determination of a dispersion formula for binary mixtures is developed and applied. The electronic polarizability of bovine serum albumin is determined and is found to be almost independent of the solvent and concentration. Hence it follows that the electronic polarizability of bovine serum albumin is almost independent of the molecular environment.
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  • 57
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    Biopolymers 13 (1974) 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 58
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    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The dielectric absorption of poly-DL-phenylalanine and poly-γ-benzyl-L-aspartate (PLAB) was measured in very dilute solutions to determine the type of molecular association and to locate the helix-coil transition. Both polypeptides were present as associated helices in chloroform. The mode of aggregation, which was determined by measuring the dipole moment and the critical frequency, did not depend on the polarity of the side chain but rather on that of the solvent.In both polymers, the dissociation of the aggregates in chloroform was observed on addition of small amounts of dichloroacetic acid; further addition of the acid lead to the helix-coil transition.No second region of dielectric absorption that might be related to the kinetics of the transition was found during the helix-coil transition of PBLA.
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  • 59
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    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Molecular conformation of some polysaccharides in aqueous solution in evidenced by changes in the optical rotation and in the elution pattern of gel filtration. The changes in the specific rotation against that in water are expressed as a molar conformational value [K]D21: -495° for colominic acid in 1.0 N NaOH solution, and -180° for hyaluronate (HA), +85° for corneal keratin sulfate, and +234° for amylose in 8 M urea solution. The gel filtration of amylose and HA dissolved in 8 M urea solution shows elution patterns distinctly different from those dissolved in water. The phenomena are attributable to a molecular conformational transition of polysaccharide molecules in aqueous solution.
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  • 60
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    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A previously described procedure for simultaneous optimization of bond lengths and angles was used to test different models for mannan I. Potential hydrogen bonds and the glycosidic angle were included in the optimization. A conformational model with bifurcated intramolecular hydrogen bonds of the type observed in the methyl cellobioside methanol complex showed the best agreement with available exprerimental data. The coordinates of this model were provided by computer calculations. The available X-ray data, however, were not sufficient for selecting this model; rather, ir data were necessary to furnish the needed information. The different conformational models tested all showed an almost constant virtual bond length O(1)-O(4) of the β-pyranose residue. This was in contrast to the previously obtained results for the α-pyranose residues.
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  • 61
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    Biopolymers 13 (1974), S. 1187-1193 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The longitudinal proton magnetic relaxation times T1 were measured for ferri (met)-and carbonmonoxy-bovine haemoglobin and equine myoglobin in 0.1 M KH2PO4 aqueous solutions near pH 6 at 5°C and 35°C from 1.5- to 60-MHz Larmor frequencies. It is concluded that the correlation time τC for the dipole-dipole interaction of electron and nuclear spins is in fact the electron (ferric) spin relaxation time τS being close to 1.5 × 10-10 sec for both metHb and metMb at 5°C. At 35°C the paramagnetic relaxation rates are not determined solely by the relaxation of protons exchanging from the haem pocket with bulk solvent. Hence, τC at 35°C cannot be calculated from the dispersion data obtained at this temperature. The relevance of this for the determination of interspin distances r is discussed.
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  • 62
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    Biopolymers 13 (1974), S. 1213-1222 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The orientation of naphthalene rings in the side chain of poly-γ-1-naphthylmethyl-L-glutamate (PNLG) in mixed solvents of dichoroethane (DCE) and hexafluoroisopropanol (HFIP) has been studied together with its conformation by infrared, circular dichroism, and fluorescence spectra. The CD pattern of PNLG varies with the solvent composition while it maintains the α-helical conformation. The fluorescence spectra of PNLG in solution show excimer emission of the naphthalene chromophores. The ratio of intensity of the excimer emission to that of the normal fluorescence decreases as the HFIP component in the solvent increases. It is suggested that the naphthalene rings in the side chain of α-helical PNLG are more rigidly orientated in the solvents of higher HFIP ratio.
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  • 63
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    Biopolymers 13 (1974), S. 1269-1290 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The near ultraviolet chiroptical properties of L-cystine conformational isomers are examined on a static, “one-electron” model in which the disulfide moiety is the chromophoric group and the atoms of the L-alanyl fragments are treated as perturbers. The zeroth order chromophoric wave functions are calculated on a semiempirical molecular orbital model in which excited states are constructed in the virtual orbital-configuration interaction approximation. The perturbing environment is represented by point charges located at the atomic centers of the L-alanyl fragments. Chromophore-perturber interactions are expressed as charge-multipole moments with only the charge-dipole and charged-quadrupole terms being retained in the calculations. Vicinal contributions to the rotatory strengths of the five lowest energy disulfide transitions are computed for 30 conformational isomers of the L-cystine dizwitterion. The results provide support for the view that vicinal or peripheral effects can account entirely for the observed near ultraviolet (λ 〉 230 nm) chiroptical properties of L-cystine and its derivatives and that these properties are diagnostic of conformational features external to the disulfide moiety.
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  • 64
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    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The circular dichroic spectrum of poly(Tyr,TyrBr) in trimethylphosphate from 180 to 320 nm is found to be a linear function of composition from 25 to 68% TyrBr, and the spectra of the Tyr and TyrBr components are resolved. For Tyr the resolved spectrum is in agreement with available CD spectra of poly(Tyr). These results are interpreted to mean that the side-chain conformations of Tyr and TyrBr are not affected by the other component, and that for Tyr the conformation is the same as in poly(Tyr). It is also concluded that poly(Tyr,TyrBr) is helical with the same helix sense as poly(Tyr). The restrictions on the side-chain conformations are discussed, and it is suggested that the Tyr and TyrBr side chains are all in the same χ1 region.
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  • 65
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    Biopolymers 13 (1974), S. 1377-1389 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The circular dichroism of double-helical nucleic acids was calculated as a function of geometry using the theory of Tinoco and Johnson. This theory does not include contributions of near ultraviolet transitions that are not π → π* in nature. The calculated circular dichroism shows a strong dependence on the distance of base pairs from the helix axis and the tilt of the base pair. Smaller dependences are predicted for the propeller-like twist of a base pair and for variation of the angular increment per base pair. Moderately good agreement between calculated and many experimentally observed spectra could be generated.
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  • 66
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    Biopolymers 13 (1974), S. 1427-1434 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The maximum at 220 nm in the circular dichroism spectrum of native collagen solution changed to a negative value after heat denaturation or collagenase hydrolysis. The enzyme induced rate of CD change at 220 nm was shown to be first order in collagen concentration. The specific rate constant k is actually a combined rate constant kfast and kslow in which the ratio kf/ks is 4.1. The initial rates were linear with respect to enzyme concentration, and the Km was found to be 5.5 × 10-7 M. The rate of ultraviolet hyperchromicity at 220 nm on collagen hydrolysis was determined. The kfast was the same as that obtained by CD. The kf/ks ratio was 4.6. Both methods may be readily used to assay for collagenase activity.
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  • 67
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    Biopolymers 13 (1974), S. 1461-1482 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The crystal and molecular structure of the complex of amylose with dimethyl sulfoxide has been studied by a combination of stereochemical analysis, potential energy, and X-ray diffraction methods. The complex crystallizes in a pseudotetragonal unit cell with a = b = 19.17 Å and c (fiber axis) = 24.39 Å, with two antiparallel chains per unit cell and space group P212121. The amylose chain is a left-handed 61(1.355) helix with three turns per crystallographic repeat. The O(6) rotational position is approximately gt. Dimethyl sulfoxide is located inside the helix with one DMSO molecule for every three glucose residues. An additional four DMSO molecules and eight water molecules each are located in the large interstices between chains, and it is the interaction of these molecules with the helix that results in the pseudotetragonal chain packing. The interstitial DMSO is the source of the previously reported additional layer lines, which are not consistent with the 8.13-Å amylose repeat distance. The final R factor for the layers with amylose contribution to the structure factors was 0.29, while the overall R factor was 0.35. The stereochemical packing analysis provided suitable phasing models for the subsequent X-ray refinement.
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  • 68
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    Biopolymers 13 (1974), S. 1549-1556 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Dielectric dispersion of DNA was studied in the frequency range 100 Hz-100 kHz at four different temperatures (6-30°C). The dielectric increment ε0-ε∞ increased with the rise of temperature. The relaxation time, on the other hand, decreased. Both the increase in dielectric increment and the decrease in relaxation time could not be explained on the basis of the counterion polarization theory. Dipole moment was estimated from Kirkwood theory. It was found to decrease systematically with temperature. Even at 0°C there was a dipole moment of 104D.
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  • 69
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    Biopolymers 13 (1974), S. 1573-1605 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The dependence of Förster long-range resonance energy transfer efficiency on the orientational freedom of donor D and acceptor A molecules attached to a macromolecular substrate is examined. The usefulness of polarized emission measurements in determining the mutual orientation as well as the degree of orientational freedom of D and A and thereby deriving maximum and minimum values for the D-A separation from the transfer efficieny is demonstrated.
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  • 70
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    Biopolymers 13 (1974), S. 1653-1659 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Five human hemoglobins whose genetic relationship to one another involves one set of alleles, hemoglobins CC, SC, SS, AS, and AA, were studied in the deoxygenated form within a McBain gravimetric sorption system at 28°C. Water was used as the adsorbate at relative vapor pressures ranging from 0.11 to 0.39. The results of this study are reported and compared with a similar study on the ferric form of these same five hemoglobins. For both forms of hemoglobin studied, a slight increase was measured in the number of polar sites accessible to H2O vapor for those genotypes in which the substituent in the sixth position from the N terminus of the two β chains had a positively charged side chain, i.e., hemoglobin C. Those samples containing hemoglobin S (less soluble in the deoxy form) show an increased BET monolayer coverage in the deoxy as compared to the ferric form. For each genotype, the deoxygenated form had more polar sites accessible to H2O vapor than its ferric form. This finding is in agreement with an increase in the size of the central cavity as shown by crystallographic analysis.
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  • 71
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    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Thermal denaturation of direct-mixed and reconstituted polylysine-DNA complexes in 2.5 × 10-4 M EDTA, pH 8.0 and various concentrations of NaCl has been studied. For both complexes, increasing ionic strength of the solution raises Tm, the melting temperature of free base pairs. The linear dependence of Tm on log Na+ indicates that the concept of electrostatic shielding on phosphate lattice of an infinitely long pure DNA by Na+ can be applied to short free DNA segments in a nucleoprotein. For a direct-mixed polylysine-DNA complex, the melting temperature of bound base pairs Tm′ remains constant at various ionic strengths. On the other hand, the Tm′ in a reconstituted polylysine-DNA complex is shifted to lower temperature at higher ionic strength. This phenomenon occurs for reconstituted complex with long polylysine of one thousand residues or short polylysine of one hundred residues. It is shown that such a decrease of Tm′ is not due to a reduction of coupling melting between free and bound regions in a complex when the ionic strength is raised. It is also not due to intermolecular or intramolecular change from a reconstituted to a direct-mixed complex. It is suggested that this phenomenon is due to structural change on polylysine-bound regions by ionic strength. It is suggested further that Na+ may replace water molecules and bind polylysine-bound regions in a reconstituted complex. Such a dehydration effect destabilizes these regions and lowers Tm′. This explanation is supported by circular dichroism (CD) results.
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  • 72
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    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Theoretical values are reported for the specific conductivity, charge carrier mobilities, and delocalized free paths in β-polyglycine using the parallel-chain pleated sheet conformation and the all-valence electron energy band structure obtained previously. The mobilities are found to be much larger along the polypeptide backbones than in the direction of the hydrogen bonds. This indicates a mechanism of conduction in which the former path is favored for electronic delocalization. The preexponential factor of the specific conductivity obtained theoretically agrees well with experimental results.
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  • 73
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    Biopolymers 13 (1974), S. 1747-1750 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The circular dichroism of a series of L-methionine oligopeptides [BOC-(Met)n-OMe] was examined in trifluoroethanol and hexafluoroacetone sesquihydrate. The results indicate that the trimer through the hexamer exists predominantly in disordered conformations in these solvents. An abrupt change in the CD pattern at the heptamer in trifluoroethanol suggests that L-methionine oligopeptides begin forming helices at this chain length.
    Additional Material: 3 Ill.
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  • 74
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    Biopolymers 13 (1974), S. 1767-1776 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The spin-lattice relaxation time (T1) of water adsorbed on collagen fibers was determined at six frequencies and temperatures varying from 25° to -80°C. Care was taken to eliminate the contributions to the signal of protons other than those in the adsorbed water. Quantitative calculations were made on T1 and the results were compared with the experimental data. It is suggested that a maximum of about 0.50-0.55 g water per g collagen forms a hydration layer, which cannot be frozen down to -90°C and exhibits a distribution of motional correlation times. For collagen samples containing a larger quantity of adsorbed water, the additional water molecules behave like ordinary isotropic water, having a single correlation time and a freezing temperature of about -10°C.
    Additional Material: 5 Ill.
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  • 75
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    New York : Wiley-Blackwell
    Biopolymers 13 (1974), S. 1821-1845 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Laser Raman spectra of the trinucleoside diphoshate ApApA and dinucleoside phosphates ApU, UpA, GpC, CpG, and GpU are reported and discussed. Assignments of conformationally sensitive frequencies are-facilitated by comparison with spectra reported here of poly(rA), poly(rC), and poly(rU) in deuterium oxide solutions. The significant spectral differences between ApU and UpA, and between GpC and CpG, reveal that the sequence isomers have nonidentical conformations in aqueous solution. In UpA at low temperature the bases are stacked and the backbone conformation is similar to that found in ordered polynucleotide structures and RNA. In ApU no base stacking can be detected and the backbone conformation differs from that found in UpA, both in the orientation of phosphodiester linkages and in the internal conformation of ribose. At the conditions employed neither ApU nor UpA exhibits base pairing in aqueous solutions. In both GpC and CpG the bases are stacked and the phosphodiester conformations are similar to those encountered for UpA and RNA. However, major differences between spectra of GpC and CpG indicate that the geometries of stacking and ribosyl conformations are different. In GpC the Raman data favor the formation of hydrogen bonded dimers containing GC pairs. Protonation of C in GpC is sufficient to eliminate the ordered conformation detected by Raman spectroscopy. Despite the ordered backbone conformation evident in GpU, this dinucleoside apparently contains neither stacked nor hydrogen bonded bases at the conditions employed here. The Raman data also confirm the stacking interactions in ApApA, poly(rA), and poly(rC) but suggest that the backbone conformation in poly(rC) differs qualitatively from that found in most ordered polynucleotide structures and is thermally more stable. The present results demonstrate the sensitivity of the Raman technique to sequence-related structural differences in oligonucleotides and provide additional spectra-structure correlations for future conformational studies of RNA by laser Raman spectroscopy.
    Additional Material: 11 Ill.
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  • 76
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    New York : Wiley-Blackwell
    Biopolymers 13 (1974) 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 77
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The 300-MHz proton nmr spectra (between 11 and 14 ppm) of a series of double-stranded deoxy oligonucleotides of known sequence have been recorded in H2O solution. These resonances have been assigned to the G—N1H and T—N3H protons of specific base pairs from an evaluation of the temperature dependence of the ring NH linewidths and from the selective ring NH chemical shift changes on actinomycin-D binding. The deoxy oligonucleotides exist predominantly in the DNA-B conformation as evaluated from antibiotic binding studies. Ring-current calculations have been utilized to evaluate the up-field shifts of the G—N1H and T—N3H protons in Watson-Crick base pairs due to the ring currents from the pyrimidine and purine rings of nearest neighbor base pairs in regular DNA-B- and RNA-A-type helices. The perturbations on these up-field ring-current contributions that arise from twisting and tilting a base pair adjacent to the ring NH under study have been evaluated and found to change the calculated chemical shift by ±0.6 ppm for twist and tilt distortions of 〈30°C in a single adjacent base pair. A knowledge of the experimentally assigned ring NH chemical shifts of specific base pairs in known sequences of double-stranded deoxy oligonucleotides coupled with the ring-current tables for the DNA-B helical structure permit the assignment of 13.6 ± 0.1 ppm and 14.6 ± 0.2 ppm for the G—N1H proton of an isolated GC base pair and the T—N3H proton of an isolated AT base pair, respectively.
    Additional Material: 8 Ill.
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  • 78
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    Weinheim : Wiley-Blackwell
    Chemie in unserer Zeit 8 (1974), S. A4 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 79
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    Chemie in unserer Zeit 8 (1974), S. 65-71 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 8 Ill.
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  • 80
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    Chemie in unserer Zeit 8 (1974), S. A40 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 81
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    Chemie in unserer Zeit 8 (1974), S. 112-120 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 15 Ill.
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  • 82
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 83
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    Weinheim : Wiley-Blackwell
    Chemie in unserer Zeit 8 (1974) 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 84
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    Chemie in unserer Zeit 8 (1974), S. 159-160 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 85
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    Chemie in unserer Zeit 8 (1974), S. 192-193 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 86
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    Chemie in unserer Zeit 8 (1974), S. A78 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 87
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    Chemie in unserer Zeit 8 (1974), S. 121-123 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 2 Ill.
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  • 88
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 89
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    Chemie in unserer Zeit 8 (1974), S. 129-134 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 7 Ill.
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  • 90
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    Chemie in unserer Zeit 8 (1974), S. A66 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 91
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    Chemie in unserer Zeit 8 (1974), S. 177-183 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 12 Ill.
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  • 92
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    Weinheim : Wiley-Blackwell
    Chemie in unserer Zeit 8 (1974) 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 93
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    Chemie in unserer Zeit 8 (1974), S. 31-32 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 94
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
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  • 95
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    New York : Wiley-Blackwell
    Biopolymers 13 (1974), S. 29-61 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: This paper describes the first experimental application of fluorescence correlation spectroscopy, a new method for determining chemical kinetic constants and diffusion coefficients. These quantities are measured by observing the time behaviour of the tiny concentration fluctuations which occur spontaneously in the reaction system even when it is in equilibrium. The equilibrium of the system is not disturbed during the experiment. The diffusion coefficients and chemical rate constants which determine the average time behaviour of these spontaneous fluctuations are the same as those sought by more conventional methods including temperature-jump or other perturbation techniques. The experiment consists essentially in measuring the variation with time of the number of molecules of specified reactants in a defined open volume of solution. The concentration of a reactant is measured by its fluorescence; the sample volume is defined by a focused laser beam which excites the fluorescence. The fluorescent emission fluctuates in proportion with the changes in the number of fluorescent molecules as they diffuse into and out of the sample volume and as they are created or eliminated by the chemical reactions. The number of these reactant molecules must be small to permit detection of the concentration fluctuations. Hence the sample volume is small (10-8 ml) and the concentration of the solutes is low (∼ 10-9 M). We have applied this technique to the study of two prototype systems: the simple example of pure diffusion of a single fluorescent species, rhodamine 6G, and the more interesting but more challenging example of the reaction of macromolecular DNA with the drug ethidium bromide to form a fluorescent complex. The increase of the fluorescence of the ethidium bromide upon formation of the complex permits the observation of the decay of concentration fluctuations via the chemical reaction and consequently the determination of chemical rate constants.
    Additional Material: 10 Ill.
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  • 96
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    New York : Wiley-Blackwell
    Biopolymers 13 (1974), S. 129-138 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The D and L copolymerizations of N-carboxy γ-benzyl glutamate anhydride (NCA) were carried out in a homogeneous solution with various D/L ratios, initiated by either n-butylamine or sodium methoxide, and were followed directly by circular dichroism (CD) to observe the behavior of the secondary structure of growing polymer molecules. In the n-butylamine system, the difference of the helical content between the righthanded and the lefthanded (Δα-helix) gradually increased as the polymerization proceeded, while in the sodium methoxide system, the Δα-helix had a tendency to decrease during the later stages of the polymerization. These results suggest a difference of the power of stereo-selection of monomer antipodes by the growing chain end between these systems, the stereoselectivity by the growing chain end in the sodium methoxide system being higher than that in the n-butylamine system.
    Additional Material: 6 Ill.
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  • 97
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    New York : Wiley-Blackwell
    Biopolymers 13 (1974), S. 169-183 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The circular dichroism of Ac-Ala-NHMe, cyclo(-Ala-Ala-), Ac-Ala-OMe, Ac-Ala-Ala-OMe, and Ac-Ala-Ala-Ala-OMe has been measured in water and in aqueous salt solutions as a function of temperature. Only cyclo(-Ala-Ala-) exhibits circular dichroism which is independent of temperature. Each of the linear derivatives of L-alanine exhibits a positive circular dichroism in the range 208-218 nm at 15°C in water. Heating reduces the intensity of the positive circular dichroism, and only Ac-Ala-OMe retains positive circular dichroism at 75°C in water. Isothermal addition of salts produces changes in the circular dichroism of linear derivatives of L-alanine which resemble those seen on heating. The relative effectiveness of the salts tested, at a concentration of 4M, is LiCl ≤ KCl = NaCl 〈 MgCl2 ≤ CaCl2 ≤ NaClO4. The circular dichroism of cyclo(-Ala-Ala-) is also affected by the salts. Extrapolation of the results obtained with Ac-Ala-OMe, Ac-Ala-Ala-OMe, and Ac-Ala-Ala-Ala-OMe to a long polypeptide with a -CH2R side chain in the L-configuration leads to the conclusion that this polypeptide should exhibit a temperature-dependent salt-sensitive positive circular dichroism between 208 and 218 nm when it exists as a statstical coil.
    Additional Material: 5 Ill.
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  • 98
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    New York : Wiley-Blackwell
    Biopolymers 13 (1974), S. 227-235 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The heats of the conformational conversion ΔHc of polyuridylic acid (polyU) in the presence of various cations were determined using a differential scanning microcalorimeter and an isothermal mixing calorimeter. The first method yields a value of ΔHc = 5.2 kcal/mol of base pairs, the second one a value of ΔHc = 6.4 kcal/mol of base pairs. Knowing ΔHc and the maximal slope of the degree of conversion, which was determined from the temperature dependence of the absorbance of polyU solutions at 260 nm, we were able to evaluate the parameter of cooperativity σ. From the relation σ = eΔFstack/RT the stacking free energy ΔFstack could be calculated. Dividing the apparent enthalpy of conversion by the calorimetrically measured enthalpy yields the cooperative length N0 of one helical segment at the midpoint of conversion. The results show that the type of the cation has no influence on the magnitude of ΔHc, whereas the cooperativity of polyU is influenced by the various cations.
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  • 99
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    New York : Wiley-Blackwell
    Biopolymers 13 (1974), S. 861-862 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 100
    Electronic Resource
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    New York : Wiley-Blackwell
    Biopolymers 13 (1974), S. 903-908 
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Clipped autocorrelation functions for the fluctuating intensity of light scattered from dilute solutions of poly-L-lysine in 0.2 M NaBr have been observed over a wide range of time scales down to 1.83 μsec, and over a wide range of pH. Two poly-L-lysine samples of molecular weight 35,000 and 200,000, respectively, were compared to determine the extent of draining of the so-called random-coil configuration. The observed correlation functions were in general very satisfactorily represented by simple translation diffusion of single molecules. At high pH ∼10-10.4, there was evidence of considerable aggregation, but no indication whatsoever of any unusually fast component, as reported recently by Jamieson et al. for poly-L-lysine HBr in the absence of salt.A 30% rise in the diffusion coefficient to a peak at pH 10 parallels the previously reported dip in the intrinsic viscosity, and is interpreted in terms of a more compact structure in that pH region between the coil and helix extremes.
    Additional Material: 3 Ill.
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