STEROLS OF PORPHYRIDIUM(RHODOPHYTA)1,2

Species of the unicellular Porphyridium have been examined for their sterol content. Clones of 4 species maintained in axenic, chemically-defined culture were analyzed—these included P. sordidum Geitler, P. purpureum (Bory) Ross, P. aerugineum Geitler and P. violaceum Kormnann(P. griseum Geitler was not available to use for examination). The major sterol was 22-dehydrocholesterol in all except P. aerugineum in which there was a mixture of this sterol, cholesterol and higher sterols. Traces of C₂₈ and C₂₉ sterols were detected in most instances as well.     

Euplotes Cytoskeleton: Tubulin and Microtubular Systems in Interphase1,2

The Euplotes cytoskeleton obtained by treatment with Triton X-100 in a microtubule-stabilizing buffer has been studied by electron microscopy and SDS-polyacrylamide electrophoresis. An antiserum has been prepared using the tubulin band from preparative gels of Euplotes cytoskeletons as an antigen. This antiserum reacts with the tubulin from different ciliated protozoa but fails to recognize the vertebrate tubulin by immunoblotting. Immunoblotting studies have demonstrated a slower electrophoretic mobility for the α-tubulin of Euplotes and Oxytricha than that of Paramecium and Tetrahymena. A cytoplasmic microtubular network in Euplotes has been revealed by indirect immunoftuorescence using both an anti-α-tubulin monoclonal antibody directed against chick brain tubulin and an antiserum raised against the tubulin of Euplotes.     

COMPETITIVE EXCLUSION AMONG THREE PLANKTONIC BLUE-GREEN ALGAL SPECIES1,2

Although domination of 1 species in an algal community has often been attributed to toxic effects, nutrient exhaustion can also lead to competitive exclusion among algal species. The bacteria-associated blue-green algae Microcystis aeruginosa, Nostoc muscorum, and Phormidium foveolarum were cultivated in Zehnder-Gorham's medium No. 11 individually and in their combinations. The growth rates of Microcystis and of Nostoc were reduced by the presence of Phormidium. This detrimental effect was apparently not due to formation of any toxic or allelopathic matter. It was caused by the reduction of iron and trace metal concentrations in the medium by Phormidium to levels too low for normal growth of Microcystis and Nostoc. Work with the axenic forms of the 3 algal species also indicated a lowering of iron and trace element concentrations by Phormidium to levels not sufficient for sustained growth of Microcystis and Nostoc. Thus, bacteria played no significant role in this example of competitive exclusion.     

Oxidative Cleavages of 1,2-Diaminosugars and their Significance in the Mechanism of the Amino-carbonyl Reactions

N-Benzyl-(l-deoxy-l-p-toluidino)-D-fructosylamine gives extensive browning on heating in methanol or ethyl acetate especially in the present acid, and from the browned solutions N-benzyl-D-arabinonamide has been isolated. Under stream of oxygen the browning is less extensive but the yield of D-arabinonamide is increased; under nitrogen the browning is slight and unchanged diaminosugar has been recovered. In the presence of an excess of benzyl-amine, benzylammonium D-arabinonate has been isolated. N-Cyelohexyl- (2-cyclohexylamino-2-deoxy) -D-glucosylamine on heating with acid in stream of oxygen gives small amounts of N, N-bis (cyclohexyl) -oxaldiamide, dicyclohexyliminoglyoxal and cyclohexylammonium D-arabinonate as the oxidation products. The mechanism of the oxidative cleavages has been explained in terms of the high reducing ability of the enediamine structures which are formed by rearrangement of 1,2-diaminosugars; in the case of N-cyclohexyl- (2-cyclohexylamino-2-deoxy) -D-glucosylamine dehydrogenation of the enediamine structure to diiminoglucosone should be involved. The findings are applied to explain the fragmentation of sugar molecules in the amino-carbonyl reactions.     

The Surface of the African Trypanosomes1,2

The African trypanosomes bear on the outside of their cell membrane a single 10–15 nm thick coat of a glycoprotein. This glycoprotein may differ in structure in the predominant populations of parasitemic waves found in relapsing infections. Variant Specific Glycoprotein (VSG) range in MW between 53,000–63,000 d and may have variable amounts of carbohydrate attached at one, two, or several loci. Such differences in carbohydrate content may account in part for their range in molecular size. Approximately 30 C-terminal residues demonstrate isotypy; i.e. these regions fall into classes having similar amino acid sequence. Modest homology has been demonstrated in two VSGs of T. congolense arising in relapsing infections although comparison of many VSG show little or no obvious homology. More recently, lipid-associated forms of VSG have been described and it is believed that these forms may be transmembrane proteins. Different VSGs appear to have different amounts of the primary sequence which have alpha-helix-forming potential. In some VSG, in excess of 80% of the structure is helical as judged by both Chou-Fasman calculations and by circular dichroism. This raises the possibility that different VSG may have different folding patterns. The arrangement of VSG on the trypanosome surface probably places the basic amino acid-rich carbohydrate-bearing C-terminus of the polypeptide chain close to the membrane. There is some protein-protein association between VSGs for which (in T. evansi) the C-terminal tail is not required. The importance of VSG structure lies not only in the fact that the molecule mediates the phenomenon of antigenic variation but also in the recent observation that VSG may act on the cellular immune system to suppress the humoral immune responses of the host.     

Absolute Configuration of 1,2-Epoxy Menthyl Acetate Isolated from a New Chemical Strain of M. rotundifolia

The effect of combining a cyclopropyl group with a benzo- 1,3-dioxole group on their synergistic property has been studied by synthesizing a large number of such compounds. The factor of synergism, R_m values and spectroscopic data of these compounds is reported.     

Identification of a homogentisate-1,2-dioxygenase gene in the fungus Exophiala lecanii-corni: analysis and implications

Exophiala lecanii-corni is a dimorphic fungus capable of degrading several volatile organic compounds (VOCs) including ethylbenzene, which has been classified as a hazardous air pollutant by the Environmental Protection Agency. In contrast to bacterial species, little is known about the mechanisms of fungal degradation of VOCs. The results described herein suggest a potential pathway for ethylbenzene degradation in E. lecanii-corni via styrene, phenylacetate and homogentisate. Consistent with this proposed pathway, a full-length homogentisate-1,2-dioxygenase gene (ElHDO) has been identified, cloned and sequenced. The nucleotide sequence of ElHDO consists of a 1,452-bp open reading frame encoding a protein with 484 amino acids. The expression of the gene product increases when grown on ethylbenzene, further suggesting that it could be involved in ethylbenzene degradation and may be responsible for the aromatic ring cleavage reaction. In addition, a 907-bp fragment isolated upstream from this gene shares 78% sequence identity at the amino acid level with the amino acid sequences of two fungal phenylacetate hydroxylase genes. This observation suggests that the genes responsible for ethylbenzene degradation may be clustered. This research constitutes the first step towards a better understanding of ethylbenzene degradation in E. lecanii-corni.     

SEXUALITY AND CYST FORMATION IN PACIFIC STRAINS OF THE TOXIC D1NOFLAGELLATE GONYAULAX TAMARENSIS1,2

Sexuality has been established for a culture of Gonyaulax tarnarensis Lebour (strain NEPCC–71). The addition of a thick inoculum to a nitrogen–deprived medium results in the occurrence of anisogamous sexual fusion within the first three days in the new culture. Planozygotes, large “lumpy” cells recognizable by their four flagella, may persist up to 2 wk before forming a smooth–walled, oval hypnozygote. The latter resembles cysts released asexually by ecdysis but has a slightly thicker wall. Viable cysts resembling hypnozygotes (zygotic cysts), but with reduced photosynthetic pigmentation, have been isolated from natural murine sediments in Hidden Basin, British Columbia, and a culture (strain NEPCC–254) was initiated from excysted individuals. Zygotic cysts of NEPCC–71 remained encysted in the light at 17 C for 8 wk before excysting. The presence of a ventral pen with toxicity in the latter strain indicates that the taxonomy of G. tamarensis-like organisms is still in a stale of flux and the criteria for recognition of G. excavata (Braarud) Balech as a separate species are not satisfactory as presently formulated.     

Aromatic metabolism in Rhizobium trifolii-catechol 1,2-dioxygenase

Catechol 1,2-dioxygenase has been purified 46-fold from cells of Rhizobium trifolii TA1 grown on benzoate plus glucose. The dioxygenase had a molecular weight of 107,000 and a sub-unit molecular weight of 59,000. The enzyme had a K _m of 2 M for catechol and also cleaved 4-methylcatechol. The dioxygenase contained 2 g atoms of Fe³⁺ per mole of enzyme which could be removed by treatment with 1,10-phenanthroline, resulting in a complete loss of activity; reactivation of the enzyme occurred specifically with Fe³⁺.     

REQUIREMENTS FOR THE MAINTENANCE AND GROWTH OF APHANIZOMENON FLOS-AQUAE IN CULTURE1,2

Aphanizomenon flos-aquae has been introduced into unialgal culture in a completely defined medium and maintained for more than 3 years in the colonial flake form found in nature. The color and size of the flake and general appearance of the cells are substantially improved over those found in the original material at the time of collection from Upper Klamath Lake, Klamath County, Oregon. The alga readily forms both akinetes and heterocysts in culture Bacteria-free cultures have not been obtained. A precipitate-free medium designated ASM No. 8a has been developed from the ASM of McLachlan and Gorham. Their medium was altered by modifying the concentration of certain elements and by the addition of sodium bicarbonate, a 1/25 dilution of Arnon's trace element solutions B₇ and C₁₃, hydrogen ferric ethylenediamine di-o-hydroxyphenylacetate (EDDHA), and disodium ethylene dinitrilotetra-acetate (Na₂EDTA).     

CHAIN-FORMING DIATOMS: THREE SPECIES OF CHAETOCERACEAE1,2

Bacteriastrum furcatum Shadbolt, Chaetoceros bacteriastroides Karsten and C. pseudocurvisetum Mangin are each connected in chains by fusion of silica between setae. The first two have specialized heterovalvate end cells indicating physiological control of the length of chains, but chains of C. pseudocurvisetum have usually been seen broken. There are specialized deeply lobed connecting bands in B. furcatum and C. pseudocurvisetum, allowing the setae of sibling valves to protrude outside the diameter of the chain during their formation. C. bacteriastroides and C. pseudocurvisetum are not identical; differences include presence of small setae, fusion of large setae some distance from the valves, symmetry of apertures between cells, and straight chains of C. bacteriastroides, compared with valve mantle connections with no small setae, large setae joined by crossing over, asymmetry of windows between cells, and curved chains of C. pseudocurvisetum. The evolutionary direction is hypothesized to be from Bacteriastrum to Chaetoceros through C. bacteriastroides.     

GLAUCOSPHAERA VACUOLATA,ITS ULTRASTRUCTURE AND PHYSIOLOGY1,2

Glaucosphaera vacuolata was recently rediscovered, and isolated by Dr. Richard Starr of Indiana University Physiological and ultrastructural studies have been conducted on this cyanome and are reported for the first time. Glaucosphaera has been found to have low rates of photosynthesis, respiration, and Hill activity. Unusual features noted include the presence of R-phycocyanin as the major accessory pigment, phycobilisomes, and eyeshot-like structures in the cyanelles.     

Optimal Fixing of the Bandwidth-Parameter for the Empirical Regression1,2

The estimator ?₀(x) of the regression r(x) = E (Y | × = x) from measured points (x_i, y_i), i = 1(1) n, of a continuous two-dimensional random variable (X, Y) with unknown continuous density function f(x, y) and with moments up to the second order can be made with the help of a density estimation f?₀(x, y) (see e.g. SCHMERLING and PEIL, 1980). Here f?₀(x, y) still contains free parameters (so-called band-width-parameters), the values of which have to be optimally fixed in the concrete case. This fixing can be done by using a modification of the maximum-likelihood principle including jackknife techniques. The parameter values can be also found from the estimators for r(x). Here the cross-validation principle can be applied. Some numerical aspects of these possibilities for optimally fixing the bandwidth-parameter are discussed by means of examples. If ?₀(x) is used as a smoothing operator for time series the optimal choice of the parameter values is dependent on the purpose of application of the smoothed time series. The fixing will then be done by considering the so-called filter-characteristic of ?C₀(x).     

THE FRESHWATER DIATOM CYMBELLA CURSIFORMIS NOM. NOV.1,2

The morphology, ecology, and taxonomy of a pennate diatom, Cymbella cursiformis nom, nov. (family Cymbellaceae), from Cedar Bog, Champaign County, Ohio, originally described from Tasmania by Hustedt as Cymbella cistula var. gracilis, are reported. Photographs of specimens on Hustedt's type, slide, are indistinguishable from Cedar Bog specimens. Scanning electron microscopy and light microscopy reveal that, this taxon cannot be considered a variety of Cymbella cistula (Ehr.) Kirchn.; therefore it has been raised to specific rank.     

PERIDINIUM GREGARIUM,A NEW SPECIES OF DINOFLAGELLATE1,2

A dinoflagellate has been found in tidepools on the coast of the Palos Verdes Peninsula in southern California. It is described as a new species, Peridinium gregarium. In the tidepool habitat, great numbers of the organism form cloud-like masses held together by a mucous matrix. The gregarious cloud formation seems to be shared by only one other species of dinoflagellate, Peridinium sociale (Henneguy ex Labbé) Biecheler, 1935.     

Chemical and biological studies on 1,2-dihydro-S-triazines

Summary A series of 1,2-dihydro-s-triazines has been studied inLactobacillus casei # 7469-pteroylglutamic acid systems. The active derivatives exhibit a competitive inhibition similar to that of 4-aminopteroylglutamic acid, but differ from the latter in that inhibition is not relieved by adenine or guanine, and at appropriate concentrations of inhibitor, is not reversed by excess pteroylglutamic acid. Differences in microbiological activity can be correlated with certain alterations in the structure of the molecule, maximum activity being exhibited by the 2,2-dimethyl-phenyl- and 2,2-dimethyl-m-chlorophenyl derivatives. The inhibitory effect of these compounds is reversed·by appropriate concentrations of dihydropteroylglutamic acid, N¹⁰-formylpteroylglutamic acid, synthetic and natural citrovorum factor, thymine and thymidine. The similarity in inhibition indices obtained vs the various forms of pteroylglutamic acid inLactobacillus casei bioassay systems and the correlation with those vs citrovorum factor inStreptococcus faecalis # 8043 andLeuconostoc citrovorum # 8081 bioassay systems suggests that inhibition ofLactobacillus casei is the result of interference with the utilization of citrovorum factor. Part I: J. Amer. Chem. Soc.74, 855, 1952; Amer. J. Path.28, 599, 1952; II: J. Amer. Chem. Soc. 1955, in press; III: J. Amer. Chem. Soc. 1955, in press; IV: Proc. Soc. Exp. Biol. Med.83, 733, 1953; V: Proc. Soc. Exp. Biol. Med.83, 740, 1953; VI: Proc. Soc. Exp. Biol. Med.83, 742, 1953. The experimental work for this and studies VIII, IX, and X in this series was done in part at the Laboratorium voor de Gezondheidscleer, Universiteit van Amsterdam, Nederland, which is directed by Professor DrA. Charlotte Ruys, and these four reports are based upon a thesis submitted by G.E.F. to the Faculteit der Wis-en Natuurkunde, in partial fulfiment of the requirements for the degree of Doctor of Science.     

Microbial production of the aromatic building-blocks (S)-styrene oxide and (R)-1,2-phenylethanediol from renewable resources

(S)-Styrene oxide and (R)-1,2-phenylethanediol are chiral aromatic molecular building blocks used commonly as precursors to pharmaceuticals and other specialty chemicals. Two pathways have been engineered in Escherichia coli for their individual biosynthesis directly from glucose. The novel pathways each constitute extensions of the previously engineered styrene pathway, developed by co-expressing either styrene monooxygenase (SMO) or styrene dioxygenase (SDO) to convert styrene to (S)-styrene oxide and (R)-1,2-phenylethanediol, respectively. StyAB from Pseudomonas putida S12 was determined to be the most effective SMO. SDO activity was achieved using NahAaAbAcAd of Pseudomonas sp. NCIB 9816-4, a naphthalene dioxygenase with known broad substrate specificity. Production of phenylalanine, the precursor to both pathways, was systematically enhanced through a number of mutations, most notably via deletion of tyrA and over-expression of tktA. As a result, (R)-1,2-phenylethanediol reached titers as high as 1.23 g/L, and at 1.32 g/L (S)-styrene oxide titers already approach their toxicity limit. As with other aromatics, product toxicity was strongly correlated with a model of membrane accumulation and disruption. This study additionally demonstrates that greater flux through the styrene pathway can be achieved if its toxicity is addressed, as achieved in this case by reacting styrene to less toxic products. See accompanying commentary by Brian F. Pfleger DOI: 10.1002/biot.201300251     

SYNCHRONOUS GROWTH IN THE COLONIAL EUDORINA ELEGANS (CHLOROPHYCEAE)1,2

Eudorina elegans Ehrenberg has been synchronized by growing the organisms at 32 C in an enriched medium on a 16:8 LD cycle. Nuclear division occurred during the light period and 2–4 h was required, for the population to complete the sequence of 4 or 5 divisions each cell normally undergoes. Each cell division required 2–3 min with ca. 15 min as the intermitotic time. The sequence was repealed, every 24 h as long as synchrony was maintained.     

Stereodynamics of Small 1,2-Dialkyldiaziridines

Diaziridines are very interesting representatives of organic compounds containing stereogenic nitrogen atoms. In particular, 1,2-dialkyldiaziridines show extraordinarily high stereointegrity. The lone electron pairs of the nitrogen atoms are in trans configuration, avoiding a four-electron repulsive interaction. Furthermore, the trans configuration of the substituents at the nitrogen atoms is energetically favored because of reduced steric interactions. Therefore only two stereoisomers (enantiomers) are observed. At elevated temperatures the enantiomers are interconverting because of the limited stereointegrity of the chirotopic nitrogen atoms. The enantiomerization rate constants and the activation parameters of interconversion are of great interest. Here, we investigated the stereodynamics of a set of small 1,2-dialkyldiaziridines bearing short substituents (Me, Et, iPr, tBu), using enantioselective dynamic gas chromatography (DGC). Separation of enantiomers of all compounds, including the highly volatile 1,2-dimethyldiaziridine, was achieved using heptakis(2,3-di-O-ethyl-6-O-tert-butyldimethylsilyl)-β-cyclodextrin in 50% PS086 (w/w) as chiral stationary phase in fused silica capillaries with a length of up to 50 m. Measurements at variable temperatures were performed and reaction rate constants were determined using the unified equation of chromatography implemented in the software DCXplorer. The activation barriers at room temperature for 1-(tert-butyl)-2-ethyldiaziridine, ΔG^╪_298K = 123.8 kJ mol^–1 (ΔH^╪ = 115.5 ± 2.9 kJ mol^–1, ΔS^╪ = –28 ± 1 J mol^–1 K^–1), and 1-ethyl-2-isopropyldiaziridine, ΔG^╪_298K = 124.2 kJ mol^–1 (ΔH^╪ = 113.1 ± 2.4 kJ mol^–1, ΔS^╪ = –37 ± 2 J mol^–1 K^–1), were determined, representing some of the highest values observed for nitrogen inversion in diaziridines. Chirality 00:000–000, 2013. © 2013 Wiley Periodicals, Inc.     

DISTRIBUTION AND TAXONOMIC REAPPRAISAL OF CTENOCLADUS (CHLOROPHYCEAE: CHAETOPHORALES)1,2

The distribution of the rare filamentous green alga Ctenocladus Borzi was examined on a world-wide basis. All the collection sites are restricted to specific inland habitats. Most of these locations are in arid regions of North America with a few scattered sites in Peru, Sicily, and Siberia. This alga has been referred to 2 genera, either Ctenocladus or Lochmiopsis Woronochin & Popova, for the past 45 years. Based on field observations, laboratory cultures, and herbarium material, Lochmiopsis is considered synonymous with Ctenocladus. The response of vegetative cell dimensions to seasonal changes (i.e., osmotic potential and temperature) in 3 saline habitats in British Columbia was also investigated. Results from the study, along with laboratory dilutions of natural saline waters, showed that cell dimensions arc not valid criteria for separating species of Ctenocladus as proposed by some authors. Consequently Ctenocladus is considered a monotypic genus with physiological variants responding to seasonal environmental conditions.