Effect of the support matrix on biotransformation of benzaldehyde to benzyl alcohol by yeast cells in aqueous and aqueous-organic two phase systems

Summary Biotransformation of benzaldehyde to benzyl alcohol bySaccharomyces cerevisiae immobilized in different support matrices was investigated. Polymers with intrinsic hydrophobic and/or hydrophilic nature as well as mixed hydrophobic and hydrophilic supports were examined both in aqueous and bisphasic aqueous-organic systems. The hydrophobic support material ENTP-2000 or mixed silicone:alginate (50-2550-75) proved to be most suitable not only for nonconventional media but also for conventional aqueous media for production of benzyl alcohol. With ENTP-2000, catalytic activity and maximum yield were 159 mol h^–1 g^–1 dry weight catalyst and 0.89 mM, respectively, in hexane containing 2% moisture. Corresponding values in aqueous media were 246 mol h^–1 g^–1 dry weight catalyst and 1.53 mM. With 5050 silicone:alginate, catalytic activity and maximum yield were 177 mol h^–1 g^–1 dry weight catalyst and 1.18 mM, respectively, in hexane containing 2% moisture. Corresponding values in aqueous media were 192 mol h^–1 g^–1 dry weight catalyst and 0.8 mM.     

Bacterial regeneration of ammonium and phosphate as affected by the carbon:nitrogen:phosphorus ratio of organic substrates

The effect of carbonnitrogenphosphorus (CNP) ratio of organic substrates on the regeneration of ammonium and phosphate was investigated by growing natural assemblages of freshwater bacteria in mineral media supplemented with the simple organic C, N, and P sources (glucose, asparagine, and sodium glycerophosphate, respectively) to give 25 different substrate CNP ratios. Both ammonium and phosphate were regenerated when CN and NP atomic ratios of organic substrates were 101 and 161, respectively. Only ammonium was regenerated when CN and NP ratios were 101 and 10–201, respectively. On the other hand, neither ammonium nor phosphate was regenerated when CN and NP ratios were 151 and 51, respectively. In no case was phosphate alone regenerated. As bacteria were able to alter widely the CNP ratio of their biomass, the growth yield of bacteria appeared primarily dependent on the substrate carbon concentration, irrespective of a wide variation in the substrate CNP ratio.     

Cellular fatty acid composition of six fastidious,gramnegative, xylem-limited bacteria from plants

Composition of cellular fatty acids was determined for strains of fastidious, Gramnegative, xylem-limited bacteria causing or associated with Pierce's disease of grapevine, phony disease of peach, plum leaf scorch, stunt of ragweed, elm leaf scorch, and periwinkle wilt. The most abundant fatty acids were straight-chain 150, 160, 170, and 180, unsaturated 161, 181, and unsaturated 17-and 19=carbon homologs. Minor fatty acids included straightchain 120, 140, 190, and 200; an unsaturated 15-carbon homolog; hydroxy-substituted 2-OH 120, 3-OH 120, and 3-OH 140; and branched chain iso-140 and iso-200. Cyclopropane acids were not detected. Physiological age had no effect on fatty acid composition. Class analysis of data indicated relative uniformity within the group. Saturated even-carbon chains comprised 31%–42%, unsaturated acids 41%–52%, saturated odd-carbon chains 10%–18%, hydroxysubstituted chains 2%–7%, and branched-chains 1%–4% of total fatty acids. The ratio of saturated-unsaturated acids ranged from 0.8 to 1.2.     

In vitro response and pedigree analysis for somatic embryogenesis of long-day photoperiod adapted soybean

Northern-adapted soybean cultivars were screened for their ability to form somatic embryos in vitro in response to exposure to 180 M 2,4-dichlorophenoxyacetic acid in a modified Murashige & Skoog (1962) medium containing B5 (Gamborg et al. 1968) organics and 43.5 mM sucrose. The 20 cultivars formed between 1 and 7 embryos per cotyledon with between 18% and 98% of the cotyledons responding. The response was genotype dependant. Three ancestral lines, 840-7-3, A.K. Harrow and Mandarin were prominent in the genetic background of long-photoperiod adapted lines and showed a high degree of somatic embryogenesis.Abbreviations 2,4-d 2,4 dichlorophenoxyacetic acid - NAA naphthaleneacetic acid     

Structure of the majorO-glycosidic oligosaccharide of monkey erythrocyte glycophorin

Sialic acids and the majorO-glycosidic oligosaccharide of glycophorin MK from monkey (Japanese monkey,Macaca fuscata) erythrocyte membranes were characterized.N-Glycolylneuraminic acid (neu5Gc) was found as the major sialic acid, which was confirmed by gas-liquid chromatography-mass spectrometry as the trimethylsilyl methyl ester. ThreeO-glycosidic oligosaccharide units were obtained from a tryptic glycopeptide that contained all of the carbohydrate units in glycophorin MK by mild alkaline borohydride/borotritide treatment. Carbohydrate analyses of the oligosaccharides revealed that they were composed of Neu5Gc, galactose andN-acetylgalactosaminitol in the molar ratios of 111 (trisaccharide), 211 (tetrasaccharide) and 111 (pentasaccharide). The content of oligosaccharide units was estimated to be 1125 for penta-, tetra- and trisaccharide, respectively, based on the yields, the molecular weight, and the number of oligosaccharide attachment sites in the amino-acid sequence. The tetrasaccharide was the major oligosaccharide and its structure was proposed to be Neu5Gc2-3Gal1-3[Neu5Gc2-6]GalNAcol.     

Circadian effects of hydrocortisone on the blood of the newt,Notophthalmus viridescens

Summary Differential counts of the leucocytes of newts,Notophthalmus viridescens, were made at four times of day (200, 900, 1400 and 2100), 72 hours after the injection of hydrocortisone acetate (experimentais) or distilled water (controls). At all times, increases in neutrophils and decreases in lymphocytes were observed in experimentais as compared to the controls (Table 1). The increases in neutrophils in the experimental newts were most pronounced at 1400, and the decreases in the lymphocytes were greatest at 2100. The least degrees of neutrophilia and lymphopenia occurred at 900. Consequently, circadian variations in response to the hydrocortisone are indicated. The possible mechanism of mediation of the variations is discussed.Supported in part by National Science Foundation grant, GY-7661, to Sweet Briar College.     

Genetics of fertility restoration in hybrid rice

Summary The cross combination involving 14 male-sterile lines in rice, when crossed with different maintainers, showed fertility restoration in certain combinations. When F₂ segregating populations were classified based on spikelet fertility, fertility restoration was shown to be governed by 31, 9331, and 1231, due to allelic differences. This indicated that the cytosterility of the same group showed monogenic fertility restoration, whereas crossing plants belonging to different cystosterile groups showed a digenic pattern of segregation.     

Prebiotic phosphorylation of nucleosides in formamide

A possible prebiotic phosphorylation method has been investigated in which formamide served as the reaction medium. Nucleotides and nucleotide derivatives were formed when nucleosides were allowed to react with different orthophosphate, hydrogen phosphate or dihydrogen phosphate salts or with different condensed phosphate salts. The reaction products obtained from the phosphorylation of adenosine were 2, 3 and 5-AMPs, 2, 5 and 3, 5-ADPs, and 2, 3-cyclic AMP. The extent of phosphorylation in formamide exceeded 50% under favorable conditions after 15 days at 70°. The acidic dihydrogen phosphates and condensed hydrogen phosphates proved to be the best phosphorylating agents. The presence of water in the medium decreased the yield of nucleotide derivatives, but some phosphorylation of adenosine was detected using dihydrogen phosphate in formamide containing water. The phosphorylation reactions were also observed for deoxynucleosides. Little decomposition of the nucleosides was detected during the reaction time needed to form nucleotide derivatives. The facility with which phosphorylation takes place in formamide under very mild conditions may justify further studies both of prebiotic phosphorylation and synthetic phosphorylation using this solvent.This work was done while the author was on leave from the Department of Chemistry, University of Colorado, Colorado Springs, Colo, 80907, U.S.A.     

Induction threshold of the Q - mutant of bacteriophage λ in Escherichia coli

Temperature induction of bacteriophage in Escherichia coli depends on bacterial population density. The lowest rate of viability loss at the temperature threshold results in maximal gene expression of . -Infection causes bacterial cells to lose cell viability and thus decrease temperature induction efficiency. In addition, shifting-up in temperature increases the probability of progeny ; thus, the mortality of bacterial hosts increases and the expression of recombinant proteins by naked significantly decrease.     

Syntheses of 1-O-benzyl-α-glucoside and 1-O-benzyl-α-maltoside by transglycosylation of α- amylase from soluble starch in aqueous solution

Glycosides, 1-O-benzyl--glucoside (BG) and 1- O-benzyl--maltoside (BM), were synthesized from soluble starch and benzyl alcohol by transglycosylation with an -amylase in a water system. BG was mostly obtained in a reaction mixture of pH 5.0, while BM was synthesized in pH 8.0. The synthesized glycosides had -configuration linkage between sugar and benzyl alcohol. The BG was rapidly hydrolyzed to benzyl alcohol and glucose by -glucosidase. The BM was hydrolyzed to BG and glucose below pH 5.0 by the -amylase used for its synthesis but it was not hydrolyzed above pH 8.0.     

On the structure of carminic acid and carmine

Summary The chemical mechanism and histochemical significance of carmine stains are not yet understood. To determine possible effects of dye configuration on staining patterns we built models of dye molecules with the Stuart-Briegleb-type of atomic models. However, steric hindrance prevented construction of carmine according to the formula suggested by Harms. A review of recent chemical literature showed that the widely accepted formula of carminic acid is incorrect; the carboxyl group is not in the 5 but in the 7-position, and the side-chain is not a methylpentose but a hexose. Models based on the revised structural formula could be combined to 211 carminic acid-Al-Ca complexes. But formation of the central Al-O-Ca-O-Al bridge of the conventional 421 carminic acid-Al-Ca formula of carmine was still impossible. It is suggested that carmine may be a 211 compound analogous to the 211 alizarin-Al-Ca complex established by Kiel and Heertjes. Investigations of carmine were rendered difficult by wide variations in the staining properties of dye samples and the lack of data concerning the composition of various batches of carmine.     

Localization of the gene for human heart fatty acid binding protein to chromosome 1p32-1p33

Among 639 spontaneous abortions between the 8th and 14th week of gestation 342 (53.5%) revealed an abnormal karyotype. While the rate of trisomies distinctly increased with advancing maternal age, a decrease in the rate of 45,X conceptuses and polyploidies was observed among abortions from older women. The overall relation of XXXXXXYY among the tetraploidies was 1411 and that of XXXXXYXYY among the triploidies was 26 361. However, when the latter was related to maternal age, a reversal of the XXXXXY ratio of 12 in the younger to 21 in the older age groups became evident. Furthermore a decrease in the rate of paternally derived partial hydatidiform moles was found among the triploid abortion specimens from older women. From these observations we conclude that digyny plays a major role in the origin of triploidy in the increased maternal age groups, while diandry related to immaturity of oocytes and impairment of oocyte cortical function is more frequent in triploid abortions from younger women.     

Nutrient limitation in Ellis Fjord,eastern Antarctica

The occurrence of high chlorophyll-a concentrations (up to 11.6gl^–1) at shallow depths within Ellis Fjord, eastern Antarctica, during spring and summer, coincided with the development of a stratified water column. Macronutrient concentrations during periods of high chlorophyll-a levels were very low. Phosphate concentrations decreased to 0.2 M, nitrate to 0.4 M and silicate to 3.9 M. High rates of nutrient depletion early in the season can be explained by strong sea-ice algal mat development. An SiNP uptake ratio of 25.513.81 indicates a strong demand for silicate. Minimum silicate levels were below those necessary for maximum diatom growth and probably contributed to the successional shift from diatoms to phytoflagellates.     

The effect of spectral light deprivation on the spectral sensitivity of the honey bee

Summary The spectral sensitivity of spontaneous phototactic behavior was tested in short wavelength deprived bees and in control bees kept outdoors. Tests were performed with a y-maze with one branch illuminated, the other dark. The relative sensitivities for the control group were: green to blue to UV = 10.272.14. Short wavelength deprived bees show a distinct decrease in their sensitivity to short wavelengths. The relation green to blue to UV here was 10.220.27. Forager bees from outdooors, short wavelength deprived for 10 days, showed a relation of green to blue to UV of 10.261.04. Electrophysiological investigation of the compound eye using electroretinogram recordings showed no difference between deprived and control animals.     

Sex determination in Sarotherodon (tilapia)

Summary The simplest possible model of the sex determination process adding autosomal influence to a minimal number of sex chromosomes was developed to explain matings of Tilapia (Sarotherodon) species. Eighteen different genotypes, each having two autosomes (AA, Aa, or aa) and two sex chromosomes (WX, WY, WW, XY, XX or YY) involved in sex determination, are predicted by the theory. Their sex (10 males and 8 females) were determined using a series of directed graphs, showing the relative strength of the chromosome pairs, developed on the basis of Chen's sex ratio results (Chen 1969). This theoretical model predicts eight different sex ratios (01, 13, 35, 11, 97, 53, 31, 10 ); three of them are not predicted by the WXYZ theory. The greatest part of these sex ratios have been obtained experimentally in extensive series of crosses between related species of Tilapia and their hybrids, carried out by several authors. The theory succeeds in explaining all of Chen's results, including those ratios 53 and 01 seen in certain crosses but not predicted by the WXYZ theory. The importance of the autosomes is seen in comparisons of the genotype pairs (AaWY, aaWY), (AaXY, aaXY) and (AAWW, AaWW) in which the first genotype in each case is male while the second is female as proven by the sex ratio results. The members of the pair differ only in the substitution of one autosome for the other. To test the theory, experiments consisting of hormonal sex reversion and a series of crosses are proposed. Finally, theoretical and practical implications of the theory are discussed.     

A DNA sequence of Drosophila melanogaster with a differential telomeric distribution

A DNA sequence (8–19T) of 2.3 kilobase pairs (kb) of Drosophila melanogaster was localized by in situ hybridization to the extreme ends of polytene chromosomes and to the chromocenter. The relative abundance of this sequence at the ends of polytene chromosomes X2L2R3L3R is 13.41.902.7. This differential distribution is probably due to different copy numbers at the individual telomeric regions. Restriction enzyme analysis of genomic DNA shows that 8–19T sequences are interspersed with other sequences. The clone 8–19T, which contains most of this interspersed repetitive sequence, is itself not internally repetitive but has a complex sequence composition. Some of these sequences are transcribed into poly(A)⁺RNA. We suggest that the ends of Drosophila chromosomes are of a complex arrangement with some sequences common to all ends.     

Selectivity of ATP-activated GTP-dependent Ca2+-permeable channels in rat macrophage plasma membrane

Outside-out configuration of the patch clamp technique was used to test whether an intracellular application of G protein activator (GTPS) affects ATP-activated Ca²⁺-permeable channels in rat macrophages without any agonist in the bath solution. With 145 mm K⁺ (pCa 8.0) in the pipette solution, activity of channels permeable to a variety of divalent cations and Na⁺ was observed and general channel characteristics were found to be identical to those of ATP-activated ones. Absence of extracellular ATP makes it possible to avoid the influence of ATP receptor desensitization and to study the channel selectivity using a number of divalent cations (105 mm) and Na⁺ (145 mm) as the charge carriers. Permeability sequence estimated by extrapolated reversal potential measurements was: Ca²⁺ Ba²⁺ Mn²⁺ Sr²⁺ Na⁺ K⁺ = 68 30 26 10 3.5 1. Slope conductances (in pS) for permeant ions rank as follows: Ca²⁺ Sr²⁺ Na⁺ Mn²⁺ Ba²⁺ = 19 18 14 12 10. Unitary Ca²⁺ currents display a tendency to saturate with the Ca²⁺ concentration increase with apparent dissociation constant (K _d ) of 10 mm. No block of Na⁺ permeation by extracellular Ca²⁺ in millimolar range was found. The data obtained suggest that (i) activation of some G protein is sufficient to gate the channels without the ATP receptor being occupied, (ii) the ATP receptor activation results in the gating of a special channel with the properties that differ markedly from those of the receptoroperated or voltage-gated Ca²⁺-permeable channels on the other cell types.DeceasedThe authors are grateful to K. Kiselyov and A. Mamin for technical assistance. The work was supported by the Russian Basic Research Foundation, Grant N 93-04-21722 and was made possible in part by Grant N R4A000 from the International Science Foundation.     

Distribution and composition of particulate organic matter in the Ross Sea (Antarctica)

The biochemical composition and spatial distribution of particulate organic matter (POM) were studied in the Ross Sea (Antarctica) in summer 1989 to assess the quantitative role of organic carbon fractions in the cycling of organic matter in the water column. Large differences in chemical composition were observed between surface and deep layers. The results indicated that, despite large geographical differences, POM was quite homogeneous, of phytoplankton origin and mostly detrital. Different ratios were used to investigate the changes in biochemical composition of particulate organic matter in relation to the ice-melting: CN (organic carbonorganic nitrogen ratio) and C-POMPOC (sum of carbohydrate, protein and lipid carbontotal organic carbon ratio) were used to analyse the percentage of refractory organic material. PPRTPCHO (proteincarbohydrate ratio) were used to establish POM age and RNADNA ratios as a relative measure of particulate activity; POCChl a and N-PPRTChl a ratios were used to estimate the autotrophic contribution to the suspended particulate organic matter. Despite its low caloric value (5.3 Kcal g POM^–1), an high caloric content in the photic layer (1.6 Kcal m^–3 of POM and 2.5 Kcal m^–3 of POC) was found thus indicating that a large amount of food was available to higher trophic levels.     

Synthesis of aspartame precursor by solid thermolysin in organic solvent

Summary The enzymatic synthesis of a peptide compound was carried out successfully in homogeneous organic solvent.Solid Thermolysin was found to catalyze the synthetic reaction of N-benzyloxycarbonyl-L-aspartyl-L-phenylalanine methyl ester (Z-APM; a precursor of sweetner Aspartame) from N-benzyloxycarbonyl-L-aspartic acid (Z-L-Asp) and L-phenylalanine methyl ester (L-PheOMe) in a 98 percent organic medium (ethylacetatebenzenemethanolwater=5029192). The dissolution of enzyme was not observed. The optimal pH shifted to acidic side by 1.0 pH unit, compared with that in aqueous medium. The enzymatic activity of solid thermolysin with an average size of 3.4×9.5 m was determined to be 0.18 moles-product/(mg-solid)·h under the initial concentrations of L-PheOMe of 0.1M and Z-L-Asp of 0.05M, and at pH 6.0 and 40°C.     

Effect of light-dark cycles with different LD time-ratios and different LD intensity-ratios on the activity rhythm of chaffinches

Summary Chaffinches (Fringilla coelebs L.) were kept at light-dark (LD) cycles with two different light intensity ratios (ca. 500.1 and ca. 101 lux) both with 1.83 hours of twilight at the onset and end of light time. The light time/dark time ratio (LD time-ratio) was varied in the range from 420 to 222 hours.Birds at the high LD intensity-ratio (500.1 lux) increased activity time () with increasing light time from LD 420 to 222. Time of activity onset, middle, and end of activity corresponded to onset, middle, and end of light time from LD 420 to 168, but the phase angles of middle and end of activity (related to midpoint of light time and midpoint of artificial dusk respectively) became more positive (advanced) at LD 204 and 222.Birds at the lower LD intensity-ratio (101 lux) increased activity time with increasing light time from LD 816 to 204 but activity time at LD 420 was equivalent to that at LD 816. The phase angles of activity onset and middle (related to midpoint of artificial dawn and midpoint of light time, respectively) became more positive at both long (LD 204) and short (LD 420) light times.By use of different twilight regimes (linear and logarithmic changes in intensity), it was shown that end of activity was more closely related with the rate of change and/or absolute level of light intensity than was onset of activity.During January and February, the birds responded to increased LD time-ratios by an increase of activity time and by strong positive phase angle shifts which correspond to observations of activity patterns of chaffinches exposed to outdoor conditions during the spring breeding season.The free-running circadian periods () of 6 birds measured at 15 lux at the end of the experiments following LD 204 averaged 22.17 hours.From the data on precision, i. e. the period-to-period variation in the phase angles of single birds averaged over all birds of one group and condition, as well as from the number of birds not synchronized with the LD cycles, it is concluded that extreme long and short light times per 24 h represent weaker zeitgebers compared to LD cycles with medium lengths of light time. Likewise, LD cycles with low LD intensity-ratios are weaker zeitgebers than LD cycles with high LD intensity-ratios.This study was carried out while the senior author was on sabbatical leave from the University of Alaska and held a fellowship from the Alexander von Humboldt Stiftung.