On the molecular structure of chromosomes

Summary It is suggested that the construction of molecular models of the chromosome may prove to be a fruitful way of attacking the fundamental problems of cytology and genetics. Certain micelles consisting of molecules of protamine-nucleate are suggested for consideration. They are designed to interpret in molecular terms the postulates of genetics and the nature of the gene, and are constructed with special reference to the findings of the cytologists as to the capacity of the chromosomes to swell, to contract, to grow and to divide.Based upon a lecture given in the physics department of the University of Manchester on May 13, 1935. The Author is indebted to W. T.Astbuby, L. E.Bayliss, J. D.Bernal, D.Crowfoot, C.D.Darlington, P.& G.Eggleton, J.M.Gulland, L.J.Harris, D.Jordan Lloyd, M. F.Howson, W. O.Kermack, E. H.Neville, M.Polanyi and J. Z.Young for advice and criticism.     

The structure of the hypothalamic neurosecretory cells of Zoarces viviparus L. under the conditions of constant dark and light during the reproductive cycle

Summary The Structure of the hypothalamic neurosecretory cells during their activity in continuous light and dark conditions and depending on the seasonal alteration has been investigated in Zoarces viviparus L. in the level of light and electron microscopy.The depletion of AF- or AT-stainable material, of the elementary neurosecretory granules and the disappearance of smooth-surfaced vesicles occur during April and June. The accumulation of the stanable material as well as the elementary neurosecretory granules and the smooth-surfaced vesicles has been observed in the cell body in late September.An excess increase in number of the smooth-surfaced vesicles of dark-induced animals and light-induced animals kept in a black stained tank is apparent. On the other hand, the disappearance of the elementary granules and the smooth-surfaced vesicles and an enlargement in the reletive nuclear surface of the neurosecretory cells of light-induced animals which were kept in a gray stained tank in April is also evident.Taking into consideration the responsiveness of both the elementary neurosecretory granules and the smooth-surfaced vesicles in relation to the external environment as well as their topographical arrangement in the cell body the possible differences in their origin and function are discussed.Numerous studies indicate that the physical environment is in part responsible for the functional properties of the hypothalamic neurosecretory centers of various vertebrates. In the level of light microscope, it has been repeatedly shown that the neurosecretory cells are activated in the animals subjected to continuous illumination, and in those subjected to long daily photo period (Oksche et al., 1958; Fiske and Greep, 1959; Öztan and Gorbman, 1960; Satyanesan, 1965). The enlargement of the nuclei and the amount of Gomori — or aldehyde fuchsin — stainable material, as well as the enzymatic activity, were used as the criteria of cellular activity in the histological preparation. In the level of electronmicroscope it has been shown that Gomori positive material (Bargmann, 1949) is represented as aggregated elementary neurosecretory granules in the ultrathin sections of neurohypophysis (Bargmann et al,. 1957). Although the fine structure of the hypothalamic neurosecretory cells of various fishes and higher vertebrates have been studied in normal and experimental conditions (Palay, 1960; Lederis, 1962, 1964; Follenius et Porte, 1962; Follenius, 1963; Murakami, 1961–1964; Gansler, 1965; Holmes, 1965) the type of granules and their function still remains as an open question (see Bargmann, 1963; Knowles, 1965).This work was aided by a grant from Nato and the Deutsche Forschungsgemeinschaft.This paper was written as a tribute in honor of the 60th birthday of Prof. Dr. Berta Scharrer.     

The application and citation of the generic nameParmeliopsis (Lecanorales,Parmeliaceae)

The nomenclatural history of the generic nameParmeliopsis is reviewed. Its correct citation is found to beParmeliopsis (Nyl. exStizenb.)Nyl., dating from 1866, not 1869 as commonly cited, withP. placorodia (Ach.)Nyl. as holotype species. There is consequently no nomenclatural problem to the adoption ofForaminella Fricke Meyer, typified byF. ambigua (Wulfen)Fricke Meyer for theParmeliopsis species with falcate conidia. A synopsis of the nomenclature of the North American and European species of both these genera is included.     

Enzymatic production of l-phenylalanine from the racemic mixture of d,l-phenyllactate

Summary The production of l-phenylalanine from the racemate d,l-phenyllactate in an enzyme membrane reactor has been examined. In a first step the racemate is dehydrogenated to the prochiral intermediate phenylpyruvate by the enzymes d-and l-hydroxyisocaproate dehydrogenase. In a second step phenylpyruvate is reductively aminated to l-phenylalanine by l-phenylalanine dehydrogenase. Both steps are dependent on coenzyme, the first one requires NAD, the second one NADH in stoichiometric amounts; in this way the coenzyme is regenerated and only required catalytically. The coenzyme is covalently bound to polyethylene glyco-20 000 and can thus be retained in the reactor analogously to the three enzymes. In order to optimize the continuous production of l-phenylalanine from d,l-phenyllactate, models of the reaction kinetics and of the reactor system have been set up. By means of the reactor model, we can calculate the optimum ratio of the three enzymes, the optimum coenzyme concentration and the optimum phenylpyruvate concentration in the feed.In this process, at a substrate concentration of 50 mM d,l-phenyllactate we reached a spacetime-yield of 28 g l-Phe/(l*d).Abbreviations PEG polyethylene glycol - d-HicDH d-hydroxyisocaproate dehydrogenase - l-HicDH l-hydroxyisocaproate dehydrogenase - PheDH l-phenylalanine dehydrogenase - V _max maximum velocity - K _M Michaelis-Menten constant - K _l inhibition constant - R₁ reaction rate of the d-HicDH forward reaction - R₂ reaction rate of the d-HicDH reverse reaction - R₃ reaction rate of the l-HicDH forward reaction - R₄ reaction rate of the l-HicDH reverse reaction - R₅ reaction rate of the PheDH forward reaction - R₆ reaction rate of the PheDH reverse reaction - d-PLac d-phenyllactate - l-PLac l-phenyllactate - PPy phenylpyruvate - l-Phe l-phenylalanine - NH₄ ammonium - residence time     

A reconsideration of the genusRabdosiella (Lamiaceae,Nepetoideae, Ocimeae)

The two generaPlectranthus andIsodon are compared and found to be very dissimilar.Isodon ist considered to be misplaced inOcimeae subtribePlectranthinae and apparently is more closely related to subtribeHyptidinae. The disjunct genusRabdosiella is compared to these two genera and regarded to be polyphyletic. The AfricanR. calycina (Benth.)Codd is returned toPlectranthus and calledP. calycinus Benth., while the AsianR. ternifolia (D. Don)Codd is placed inIsodon sect.Pyramidium and calledI. ternifolius (D. Don)Kudo.     

The cytosolic pathway of l-malic acid synthesis in Saccharomyces cerevisiae: the role of fumarase

Saccharomyces cerevisiae accumulates l-malic acid but only minute amounts of fumaric acid. A ¹³C-nuclear magnetic resonance study following the label from glucose to l-malic acid indicates that the l-malic acid is synthesized from pyruvic acid via oxaloacetic acid. From this, and from previously published studies, we conclude that a cytosolic reductive pathway leading from pyruvic acid via oxaloacetic acid to l-malic acid is responsible for the l-malic acid production in yeast. The non-production of fumaric acid can be explained by the conclusion that, in the cell, cytosolic fumarase catalyzes the conversion of fumaric acid to l-malic acid but not the reverse. This conclusion is based on the following findings. (a) The cytosolic enzyme exhibits a 17-fold higher affinity towards fumaric acid than towards l-malic acid; the K _m for l-malic acid is very high indicating that l-malic acid is not an in vivo substrate of the enzyme. (b) Overexpression of cytosolic fumarase does not cause accumulation of fumaric acid (but rather more l-malic acid). (c) According to ¹³C NMR studies there is no interconversion of cytosolic l-malic and fumaric acids.     

On the glycocalyx,the external coat of the plasma membrane,of some secretory cells

Summary The free surface of epithelial cells of secretory organs (human placenta, lactating mammary gland of the rat, choroid plexus of man and rat) and of the accessory organs of the genital tract of the male rat is characterized by a plasmalemmal differentiation named glycocalyx or surface mucous coat. This structure is built up by filamentous or globular substructures.Two main ultrastructural types of the glyeocalyx were observed: 1) The filamentous type such as in the rat epididymis, which resembles the cat intestinal glyeocalyx (Ito, 1965) and that one of human transitional epithelium (Monis and Zambrano, 1968), and 2) The globular type, as observed in the lumen of the lactating mammary gland of the rat.Sialic acid was demonstrated histochemically in the luminal glyeocalyx of all organs studied. In addition, the glyeocalyx of acinar cells of the lactating mammary gland contains sulfate and phosphate groups which were identified by histochemical technics, using enzymatic digestion procedures, suggesting the chemical heterogeneity of this glyeocalyx.Present investigations follow the working hypothesis that the complex carbohydrates of glycocalyces become part of the product of activity of secreting cells.We thank Mr. Luis Iwakawa, Miss Silvia Falcón, Miss Elsa M. Orgnero for technical help, Miss Graciela Aliaga for secretarial assistance. Photography by Mr. H. Magnani. Dr. Hugo F. Carrer cooperated in the initial stages of this investigation.The authors acknowledge the use of the electron microscope of the Department of Pathology, Córdoba University Medical School, for which they thank Prof. E. Mosquera and Dr. E. Hliba. Dr. Hliba photographed picture number 4.     

On the validity of the genusDactylococcopsis (Cyanophyceae)

Summary The genusDactylococcopsis Hansg. 1888 (Cyanophyceae) is based on the typeD. rupestris, which was later identified as a green algae. Most of the many species described later were also placed to other groups of algae. Several authors even doubted about the existence of the genus. As, however, some species of Cyanophyceae correspond to the original generic diagnosis, the name Dactylococcopsis Hansg. ex R. et F.Chod. 1925 has been proposed as a nomen conservandum, and a new type (D. smithii R. et F.Chod.) has been defined. Further speciesD. linearis Geitl. 1935 and D.Planctonica Teil. 1942 has been unambiguously described till now.     

On the taxonomic position ofSilene thebana (Caryophyllaceae)

Silene thebana Orph. exBoiss. is transferred as a subspecies toS. fabaria (L.)Sm., resulting in the new combinationS. fabaria (L.)Sm. subsp.thebana (Orph. exBoiss.)Melzh. The chromosome count of 2n = 24 is recorded for the first time for this taxon.Dedicated to Prof. DrK. H. Rechinger on the occasion of his 80th birthday, 16th October 1986.     

Structural characteristics of the saxitoxin receptor on nerve

Summary The effects of uranyl ion (UO ₂ ²⁺ ; at low concentrations binds specifically to phosphate groups) and the cationic dye methylene blue (MB⁺; binds strongly to carboxyl groups) on saxitoxin (STX) potency in crayfish axon has been studied by means of intracellular microelectrodes. At pH 6.00±0.05 and 13.5mm Ca²⁺, addition of 10.0 m UO ₂ ²⁺ +5.0nm STX had only slightly, if any, less effect on the spike's maximum rate of rise [0.79±0.04 (viz., mean±sem) of control value] than did addition of 5.0nm STX alone (0.72±0.05). Under the same conditions of pH and Ca²⁺ concentration, 1.0mm MB⁺ had approximately the same effect: 1.0mm MB⁺+5.0nm STX, 0.76±0.03; 5.0nm STX alone, 0.70±0.04. However, at pH 7.00±0.05 and lower Ca²⁺ concentrations, 1.0mm MB⁺ significantly reduced STX potency. Using 6.0mm Ca²⁺: 1.0mm MB⁺+5.0nm STX, 0.92±0.01; 5.0nm STX alone, 0.68±0.08. Using 3.0mm Ca²⁺, the corresponding values were 0.94±0.03 and 0.67±0.04. It is concluded that: (1) In accord with previous suggestions, the ionized acidic group known to exist in the Na channel (and to which a guanidinium group of STX appears to bind) is very likely a carboxyl group and not a phosphate group. (2) The accessible part of the Na channel mouth serving as the saxitoxin receptor probably does not include phospholipid in its structure proper.     

Properties of the sugar carrier in baker’s yeast

The uptake by baker’s yeast of monosaccharides (d-arabinose,d-xylose,d-ribose,d-glucose,d-galactose) at very low concentrations (0.02–2mm) shows a distinct asymmetry, the apparent intracellular concentrations attaining values up to 85 times higher than in the medium. A practically irreversible binding inside the cell appears to be involved. The binding has an apparent activation energy of 11000 cal/mole. It is not affected by metabolic inhibitors but requires cell integrity for its occurrence. Some features of the binding suggest that the monosaccharides are partly incorporated into cell components, but whether the process is enzymic in nature remains unclear.     

Effect of the interrupted adaptation to D-methionine on the efficiency of Rhizobium meliloti strains

Summary R. meliloti strains 107-1, 111 and 152 were adapted to D-methionine in three ways: a) consecutive transfer in the presence of increasing amounts of D-methionine, b) alternate transfer between D- and L-methionine-containing media followed by final cultivation in the presence of each isomer, c) alternate transfer between D-methionine and medium 79 followed by cultivation in medium 79 or in D-methionine-medium. At the end of the experiment efficiency of the strains was ascertained by a plant test.Strain 111 lost efficiency when it was adapted consecutively to 0.125% D-methionine or alternated between D-methionine and either L-methionine or medium 79-Strain 107-1 sucessively adapted to D-methionine lost efficiency within 16 weeks. On adaptation to D-methionine alternated with L-methionine, efficiency was retained in L-methionine medium and lost in D-methionine medium. On alternate adaptation between D-methionine and medium 79, strain 107-1 lost efficiency in the D-methionine-medium but not in medium 79. Efficiency of strain 152 was lost by adaptation to 0.125% D-methionine, but it was maintained on the alternate adaptation between D-methionine and L-methionine or medium 79.     

Multivariate analysis of theSenecio nemorensis group (Compositae) in the Carpathians with a new species from the East Carpathians

Six species and subspecies of theS. nemorensis group from the Carpathians are recognized here, based on morphometric analyses (cluster and discriminant analyses). A new speciesS. ucranicus Hodálová and a new hybridS. ×dominii Hodálová=S. germanicus Wallr.×S. ucranicus Hodálová, occurring in the East Carpathians and the eastern part of the South Carpathians, are described. The vicarious nature ofS. hercynicus Herborg subsp.hercynicus, S. dacicus Hodálová etMarhold andS. ucranicus Hodálová in the Carpathians is documented. A key to theS. nemorensis group in the Carpathians as well as toS. nemorensis L. s. str. is presented.     

Chiral Selection in Oligoadenylate Formation in the Presence of a Metal ion Catalyst or Poly(U) Template

The lead ion-catalyzed oligomerization of 5′-phosphorimidazolides of D-, L- or racemic DL-adenosine (D-ImpA, L-ImpA and DL-ImpA) gave oligoadenylates up to a pentamer. The oligomers resulting from racemic ImpA were comparable in yields and length to those from chiral D- or L-ImpA. A complex mixture of homochiral and heterochiral oligomers was formed in the reaction from racemic ImpA. Total dimer product from racemic ImpA by the lead ion catalyst showed homochiral selectivity. The reaction catalyzed by uranyl ion yielded oligoadenylates up to 15mer from chiral D- or L-ImpA in over 95% yield. A complex mixture of isomeric oligoadenylates was formed from racemic DL-ImpA in the presence of uranyl ion catalyst in comparable yields to those from D- or L-ImpA. The analysis of the dimer product from DL-ImpA showed that the homochiral 2′ –5′ linked dimer was selectively formed. D-ImpA polymerized effectively on a poly(U) template, which is exclusively composed of D-uridine, yielding oligoadenylates up to a pentamer. In contrast, L-ImpA or racemic DL-ImpA polymerized far less efficiently on the poly(U) template, demonstrating that chiral selection takes place in the poly(U) template-directed oligoadenylate formation.     

On the Lichen generaPlacocarpus andPlacidiopsis (Verrucariaceae)

Verrucaria schaereri (Fr.)Nyl. is accorded generic status asPlacocarpus Trev. emend.O. Breuss. The main differences between the two genera concern the thallus anatomy and the spores which are halonate inPlacocarpus. The following new combinations are proposed:Placocarpus schaereri (Fr.)O. Breuss andPlacidiopsis cinerascens (Nyl.)O. Breuss.     

Quantification of the histochemical reaction for alkaline phosphatase activity using the indoxyl-tetranitro BT method

Summary The indoxyl—tetranitro BT method for the demonstration of alkaline phosphatase activity has been optimized and its validity for quantitative histochemistry tested. The study has been performed with model films of polyacrylamide gel incorporating homogenate of rat liver and with cryostat sections from the same livers. Addition of polyvinyl alcohol to the incubation medium greatly improved the localization of the final reaction product in cryostat sections. In polyacrylamide films, the formazan production specifically due to alkaline phosphatase was highest when using a medium containing 100mm Tris-HCl buffer, pH 9.0, 0.2–1.0mm substrate, 0.32mm 1-methoxyphenazine methosulphate, 10mm MgCl₂, 5mm sodium azide and 1mm tetranitro BT. For the incubation of cryostat sections in the presence of polyvinyl alcohol, the same medium could be used but the optimum concentrations of substrate and tetranitro BT appeared to be 1–2mm and 5mm respectively. The test minus control reaction was specific for alkaline phosphatase activity and could be inhibited completely with tetramisole. The test minus control reaction was linear with time up to 30 min with model films and up to 15 min with cryostat sections. The formazan production was also linear with the amount of homogenate incorporated in model films and with section thickness up to 18 µm and therefore, the reaction obeyed the Beer—Lambert law. Variation of the substrate concentration yielded aK _M of 0.05mm for aqueous media and aK _M of 0.55mm for polyvinyl alcohol-containing media. The inhibition with tetramisole appeared to be competitive withK _i = 0.07mm for aqueous media andK _i = 0.7mm for polyvinyl alcohol-containing media. These values indicate that the indoxyl—tetranitro BT method is considerably more sensitive than any metal salt or diazonium salt method developed so far. It is concluded that the optimized method described here is a specific, sensitive and valid quantitative histochemical method for the demonstration of alkaline phosphatase activity.     

Inhibitory action of serine on growth of bacteria of the genusBacillus on mineral synthetic media

l-Serine added to minimal synthetic media stops the growth ofBacillus subtilis, B. megaterium, B. mycoides andB. pantothenticus. All tested subspecies ofBacillus thuringiensis appear resistant to this amino acid. Serine acts bacteriostatically onB. subtilis strain B 003 and this effect depends on the concentrations of both the amino acid and the plated bacterium. Growth of serine-sensitiveBacillus strains can be restored by simultaneous addition of some other amino acids (e.g. l-threonine,l-arginine,l-aspartate orl-alanine) to the minimal media. This alleviating effect depends on the kind of amino acid. Some amino acids (e.g. l-threonine,l-tyrosine orl-tryptophan) are only effective when the serine concentration is not higher than 125 μmol/L, others (l-arginine,l-proline,l-alanine,l-aspartate orl-glutamate) are effective even when the serine concentration is as high as 500 μmol/L.     

Revision of Hyolitha from the Ordovician of Estonia

Topotype specimens of the Middle OrdovicianHyolithes acutus Eichwald, 1840, which is the type of the genus that lent its name to a family, order, class, and even phylum according to some, andH. latus Eichwald, 1860 allow that genus and those species to be firmly established on a sound, morphologic basis. In addition, preservation of the types ofHyolithes striatus Eichwald, 1860 is sufficiently good to warrant reassignment toDorsolinevitus Syssoiev, 1958. In contrast, the type ofH. insularis Eichwald, 1860 is incompletely preserved, and this species is not recognizable beyond the type material. The concept of the family Hyolithidae is revised to more closely conform to the morphology ofHyolithes, with authorship herein ascribed toSyssoiev (1958) rather than toNicholson (1872). The stratigraphic distribution of these taxa suggests thatHyolithes as defined herein first appears in the Middle Ordovician, but extends into at least the Lower Devonian, as suggested by two species from the Barrandian region of the Czech Republic. Their geographic distribution further re-enforces the notion of two distinct paleobiogeographic provinces based on hyoliths, a Mediterranean province and Baltic province, with almost no mixing of hyolith faunas during the Ordovician.      

Regulation of aspartokinase activity in the genus Beneckea and marine,luminous bacteria

Summary The pattern of allosteric regulation of aspartokinase activity was determined in species of Beneckea and in the marine, luminous bacteria. The results indicated that these organisms have at least three isofunctional aspartokinases of which the first is inhibited by l-threonine, the second is inhibited by l-lysine, and the third is unaffected by either l-threonine or l-lysine. The homoserine dehydrogenase activity is clearly separable from the l-lysine-sensitive aspartokinase, that may be associated with one (or both) of the other isofunctional aspartokinases.The species of Beneckea and luminous bacteria studied varied in the relative levels of the three isofunctional aspartokinases. B. parahaemolytica, B. alginolytica, B. pelagia, B. neptuna, B. harveyi, B. nigrapulchrituda, B. natriegens, and Photobacterium fischeri had predominant levels of the l-lysine-sensitive activity; B. campellii, P. phosphoreum, and P. mandapamensis had predominant levels of the l-threonine-sensitive activity; while in B. nereida these two activities were approximately equivalent. Species of Beneckea could be distinguished from P. fischeri on the basis of the sensitivity of their aspartokinase activities to inhibition by l-lysine. In P. fischeri 10.3×10^-5 M l-lysine was required for a 50% inhibition of the l-lysine-sensitive enzyme, while in species of Beneckea 0.5–2.7×10^-5 M l-lysine was required to achieve the same level of inhibition.Non-standard abbreviations DAHP 3-deoxy-d-arabino-heptulosonate 7-phosphate - GC guanine plus cytosine