Low Resolution Mass Spectra of Some Unsaturated δ-Lactones

Mass spectra of the δ-lactones of the following 5-hydroxy-2-enoic acids were determined: 5-hydroxyhex-2-enoic acid (I), 5-hydroxyoct-2-enoic acid (II), 5-hydroxydec-2-enoic acid (III), 5-hydroxydodec-2-enoic acid (IV), 5-hydroxy-8-methylnon-2-enoic acid (V), 5-hydroxy-6-ethyloct-2-enoic acid (VI), 5-hydroxy-5, 6, 6-trimethylhept-2-enoic acid (VII), and 5-hydroxy-5-methylnon-2-enoic acid (VIII). The following modes of fragmentation are consistent with observed m/e values, metastable peaks, and established modes of breakdown in compounds containing similar atomic groupings:—1. Loss of side chain, resulting in ions at m/e 97 for I-VI and at m/e 111 and 153 for VII and VIII (diagnostic peaks); 2. 1,4-Rupture of the ring giving an ion at m/e 68 (diagnostic peak) which loses CO to give m/e 40; 3. Loss of CO from m/e 97 fragment to give m/e 69 which breaks down further to m/e 41→m/e 39; 4. 1, 4-Rupture of m/e 111 and m/e 153 fragments to give m/e 43 and 85, further breakdown of m/e 85→57→41→39; 5. Loss of H₂O from the molecular ion providing there is a hydrogen atom on C₅ and the side chain is at least 3 carbon atoms in length, further loss of H₂O when the side chain is equal to C5 or C7; 6. Loss of CO₂ from the molecular ion of I, IV-VIII; 7. Loss of CO from all molecular ions; 8. Loss of 2×28 from the molecular ions of III, IV, V, VI; 9. Loss of (18 + 28) from the molecular ion of III, IV, V, VI; 10. Loss of 60 from the molecular ion of II, III, IV, V, VI; 11. Formation of M + 1 ion (169) of VII and VIII; 12. Formation of M + 1 ion (143) of saturated δ-octalactone and loss of H₂O from this M + 1 ion.     

Characterization of O-trimethylsilyl derivatives of d-glucose,d-galactose,and d-mannose by gas-liquid chromatography-chemical-ionization mass spectrometry with ammonia as reagent gas

The per(trimethylsilyl) ethers of d-glucose, d-galactose, and d-mannose were analyzed by g.l.c.-c.i.m.s. with ammonia as the reagent gas. C.i.m.s. gave simple fragmentation and fragment ions of high intensity in the high-mass range where the QM⁺ ion is also detected. The β-d anomers gave ions at m/e 558 showing intensities 3–12 times those of the α-d anomers. The epimers could be distinguished by differences in the intensities of the ions and by the observation that d-glucose gave a base peak at m/e 198, d-galactose at m/e 468, and d-mannose at m/e 204. The pyranose and furanose structures could be distinguished by comparing the ion intensities at m/e 198, m/e 271, m/e 361, m/e 396, and m/e 451. A similar analysis was also performed with 2-methylpropane as the reagent gas.     

Identification of 4-O-methyl-D-xylose as a constituent of the cell wall of Chlorella vulgaris

From the cell wall of a strain of Chlorella vulgaris a sugar was isolated after acid hydrolysis and was identified as 4-O-methyl-D-xylose by the following criteria: (i) mass spectroscopy of its alditol acetate revealed characteristic primary fragments with m/e 117 and m/e 261, and, when one deuterium atom was substituted at C-1, with m/e 262 instead of m/e 261; (ii) after demethylation with BCl₃, xylose was identified as its parent sugar by chromatographic methods; (iii) L-iditol: NAD 5-oxidoreductase (sorbitol dehydrogenase) catalyzed the oxidation of its alditol, but not of 4-O-methyl-L-xylitol. 4-O-Methyl-D-xylose amounted to approx. 10% of the cell walls' dry weight or 1.6% of the cells' dry weight.     

Determination of plasma testosterone by mass fragmentography using testosterone-19-d3 as an internal standard : Comparison with radioimmunoassay

Analytical procedures for the measurement of testosterone by mass fragmentography (MF) using trideuterated testosterone (testosterone-19,19,19-d₃) are described. For the calculation of plasma testosterone, peak height ratios were measured by MF performed on the molecular ions of the TFA derivative of testosterone (m/e 480) and testosterone-19,19,19-d₃ (m/e 483). The sensitivity of the method was judged from the lower limit of detection of the mass spectrometer which was at 10 pg. For the measurement of the precision, the inter- and intra-assay coefficients of variation (C.V.) were calculated by using a pooled plasma sample; they were 3.15% and 1.79%, respectively. The specificity was investigated by the use of 5α-dihydrotestosterone and the MF method was found to afford a highly selective technique. These results obtained by MF have been compared with the results obtained by a radioimmunoassay method.     

New furofurano lignans from Justicia simplex

A new 3,7-dioxabicyclo[3,3,O]octane lignan, named justisolin, and a new lignan O-glucoside, named simplexoside, were isolated from the whole plant of Justicia simplex D. Don. (Acanthaceae), collected at fruiting. The structure of the free lignan was established as 2e-(3,4-methylenedioxy-6-hydroxy)-phenyl-6e- Piperonyl-3,7-dioxabicyclo[3,3,0]octane (1) and that of the glucoside as 2e-(3-methoxy-4-O-β-d- glucopyranosyl)-phenyl-6e-piperonyl-3,7-dioxabicyclo[3,3,0]octane (2) on the basis of chemical transformation and spectral evidence. The biological functions of these and related lignans are appraised.     

Coexistence of cytoplasmic and nuclear genes for male sterility in Petunia

Summary Cytoplasmic male sterility (cms) and nuclear male sterility (nms) in Petunia were described respectively as possible autonomous and integrated states of the same genetic element by Frankel (1971). In the present study we describe genetic analysis of the interaction between the cms, the nuclear gene for male sterility (e) and the fertility restorer allele (Rf). The main findings in this study are: (1) The nuclear sterility allele can coexist in one or two dosages with the cytoplasmic male sterility elements (ste) in somatic cells or female gametes; (2) the presence of the fertility restorer allele Rf is not required for the coexistence of ste and e and (3) Rf does not interact epistatically with e, e.g., the expression of e is independent of Rf—the genotypes (S) RfRfee and (S) Rfrfee are male sterile.Contribution from the Agricultural Research Organization. The Volcani Center, Bet Dagan, Israel. 1983 series No. 846 E     

Aspects of cytotoxic lignan biosynthesis in suspension cultures of Linum nodiflorum

Plant cell suspension cultures of Linum flavum, Linum nodiflorum and Linum album have been used to characterize the growth and production of cytotoxic lignans as well as to study the biosynthesis of these lignans. A cell culture of Linum nodiflorum accumulated up to 1.7% of the cell dry weight as 6-methoxypodophyllotoxin in only nine days of cultivation. The biosynthesis of podophyllotoxin and 6-methoxypodophyllotoxin follows the formation of the first aryltetralin lignan deoxypodophyllotoxin. Hydroxylation in position 7 by deoxypodophyllotoxin 7-hydroxylase leads to podophyllotoxin. Hydroxylation in position 6 by the cytochrome P450 monooxygenase deoxypodophyllotoxin 6-hydroxylase yields -peltatin which is further methylated by S-adenosylmethionine:-peltatin 6-O-methyltransferase to -peltatin-A methylether and then hydroxylated to 6-methoxypodophyllotoxin. Both, podophyllotoxin as well as 6-methoxypodophyllotoxin are stored as glucosides in the vacuole. Certain enzymes of these transformations have been isolated and characterized from Linum cell cultures.     

Determination of an antisecretory trimethyl prostaglandin E2 analog in human plasma by combined capillary column gas chromatography—negative chemical ionization mass spectrometry

A method is described for measuring a trimethyl prostaglandin E₂ analog, TM-PGE₂, in human plasma. Trideuterated and monofluorinated analogs of TM-PGE₂ are added to plasma as internal standard and carrier, respectively. The plasma is adjusted to pH 3.0 and is extracted with a mixture of benzene—dichloromethane (9:1). The residue, following removal of the extracting solvent, is reacted consecutively with pentafluorobenzyl bromide and bistrimethylsilyltrifluoroacetamide. The excess derivatizing reagents are removed by evaporation, and an aliquot of the reconstituted residue is analyzed by capillary column gas chromatography using methane as the carrier gas. A quadrupole mass spectrometer is set to monitor in the gas chromatographic effluent the (M − C₇H₂F₅)⁻ fragmention of TM-PGE₂ (m/e 449) and trideuterated TM-PGE₂ (m/e 452) generated by methane negative chemical ionization. Quantitation of unknowns is based on a comparison of the m/e 449 to m/e 452 ion ratio in each unknown to that obtained from the analysis of control plasma spiked with known amounts of TM-PGE₂ and fixed amounts of internal standard and carrier. The sensitivity limit of the assay is approximately 100 pg ml⁻¹, which is equivalent to 1 pg injected. The assay was used to measure the concentration of TM-PGE₂ in the plasma of two subjects following a single 10 μg kg⁻¹ oral dose of the drug.     

Photosynthetic responses to salinity in two clones of Agrostis stolonifera

Abstract The response of photosynthesis, F_e, to salinity was examined in two clones of Agrostis stolonifera, one derived from a salt marsh (SM) and the other from an inland site (IL). The response of young and old leaves was examined over one week of salt treatment and changes in leaf water relations and concentrations of Na⁺, Cl^? and K⁺ ions were also monitored. Sodium⁺ and chloride^? concentrations rose much more sharply in the older leaves of both clones and F_e declined to ca. zero by Day 4. F_e in the young leaves of the SM clone was unaffected by salt whilst the young leaves of the IL clone showed a 20% reduction in F_e within 24 h of salt application. This decline was due to a decline in stomatal conductance and a reduction in the photosynthetic capacity of the mesophyll and was associated with a greater accumulation of Na⁺ and Cl^? ions and loss of K⁺ ions than in the SM clone. The significance of the salt exclusion from the young leaves of the SM clone to its growth on the salt marsh is discussed.     

Removal of organics and ions from wastewater using a hybrid system of microfiltration membrane and microorganisms

Microfiltration membranes are not able to remove organic compounds. A hybrid system of a microfiltration membrane and bacteria was designed for separation of organic compounds and ions from wastewater. Colonies of bacteria (Escherichia coli) were subjected to sedimentation on the surface of a microfiltration membrane (0.2 μm cellulose acetate) as a complementary part of the system to enhance the removal efficiency. Three selected categories of materials i.e. preservative substances, cephalosporins and ions were used to prepare the synthetic feed. The results indicate that preservatives were reduced more than 80%, cephalosporins were removed around 60% and ions decreased more than 50% in the feed solution using this hybrid system for filtration. The interaction between bacteria and chemical materials is responsible for removal of organic compounds and ions from test solutions. However the interaction decreased over time due to the limited capacity of the bacteria.     

Gas chromatographic—mass spectrometric determination of plasma 5-fluorouracil after administration of 1-hexylcarbamoyl-5-fluorouracil to dogs and humans

A simple, sensitive and specific method for determining 5-fluorouracil (5-FU) in plasma after the administration of 1-hexylcarbamoyl-5-fluorouracil (HCFU) was developed using gas chromatography—mass spectrometry. Thymine was used as the internal standard. After removal of interfering substances with chloroform, diethyl ether and Amberlite XAD-2 resin, 5-FU and thymine were extracted with 16% n-propanol in diethyl ether and methylated with trimethylanilinium hydroxide. Fragment ions at m/e 158 and 154, the molecular ion of the dimethyl derivatives of 5-FU and thymine, respectively, were used to monitor 5-FU and thymine. The sensitivity of the method is 10 ng/ml, which is sufficient to determine the 5-FU levels in plasma after the administration of therapeutic doses of HCFU to patients.     

Determination of the plasma parameters in the PF-3 facility by the methods of X-ray spectroscopy

NeIX and NeX spectra emitted by the PF-3 high-current (2 MA) plasma focus facility are measured. A numerical model describing the spectral intensities of the emission of helium- and hydrogen-like neon ions from an optically thick plasma is proposed. The electron temperature T _e and electron density n _e in the plasma of the PF-3 facility are determined by comparing the calculated and measured emission spectra of neon.     

Knots in trees – A new rich source of lignans

Recent research in our group has revealed that knots, i.e. the branch bases inside tree stems, commonly contain 5–10% (w/w) of lignans. Norway spruce (Picea abies) knots contain as much as 6–24% of lignans, with 7-hydroxymatairesinol (HMR) as the predominant (70–85%) lignan. Some other spruce species also contain HMR as the main lignan, but some spruce species have also other dominating lignans. Most fir (Abies) species contain secoisolariciresinol and lariciresinol as the main lignans. Lignans occur also in knots of pines (Pinus spp.), although in lower amounts than in spruces and firs. Scots pine (Pinus silvestris) knots were found to contain 0.4–3% of lignans with nortrachelogenin as the main lignan. Lignans have been identified also in knots of some hardwoods, although flavonoids are more abundant in hardwoods. Knots are detrimental in the manufacture of pulp and paper and should preferably be removed before pulping. This is possible using a recently developed industrially applicable process called ChipSep. Recent research has also established novel synthetic routes to several lignans, such as matairesinol, secoisolariciresinol, lariciresinol and cyclolariciresinol, starting from hydroxymatairesinol by applying fairly straight-forward chemical transformations. We conclude that wood knots in certain spruce and fir species constitute the richest known source of lignans in nature. The lignans occur in knots in free form and are easily extracted by aqueous ethanol, or even by water. Not only HMR, but also other potentially valuable lignans, could be produced in a scale of hundreds of tons per year by extraction of knots separated from wood chips at pulp and paper mills.     

New Lignans from the Flower of Forsythia koreana and Their Suppression Effect on VCAM‐1 Expression in MOVAS Cells

Six lignans including two new lignans were obtained as the principal components of the Forsythia koreana flowers via silica gel (SiO₂), octadecyl SiO₂ (ODS) as well as Sephadex LH‐20 column chromatography. In addition to two new lignans, named koreanaside A ((7R,8S,7′R,8′S)‐7,7′‐diepoxy‐5′‐hydroxy‐3,3′‐dimethoxylignan 4‐O‐β‐d ‐glucopyranoside) and koreanaside B ((7R,8S,7′S,8′R)‐7,9′‐epoxy‐9,5′,7′‐trihydroxy‐3,3′‐dimethoxylignan 4‐O‐β‐d ‐glucopyranoside), four known lignans were identified to be (+)‐phylligenin, (?)‐epipinoresinol, pinoresinol, and tinosposide A. The structures and absolute configurations of koreanasides A and B were established by means of analysis of spectroscopic data (NMR, IR, FAB‐MS, and CD), whereas the structures of known lignans were identified by comparison their NMR and MS values with those in the reported literature. Their chemical structures including configuration were established by means of analysis of spectroscopic data (NMR, IR, FAB‐MS, and CD) but also comparison of their NMR and MS values with those in the reported literature. This is the first article for isolation of six lignans of F. koreana flowers. Koreanasides A and B showed high radical scavenging activity with oxygen radical absorbance capacity (ORAC) values of 0.97 ± 0.01 and 1.02 ± 0.01, respectively. Koreanaside A also prohibited expressing VCAM‐1 in MOVAS cells with 80.5% at 25 mg/mL.     

Locomotor response of the goldfish to polarized light and itse-vector

Summary The locomotor orientation of eleven goldfish, 20–25 cm long, was monitored during periods varying between 24 hours and 8 1/2 days, to verify the response to a depolarized and polarized sky, 100 cm in diameter, and to abrupt 90 ° degree rotation of thee-vector. The monitor consisted of a cylindrical tank with 16 peripheral compartments (Fig. 1) to which the fish had free access. Entry into and exit from each compartment was electronically recorded. The distribution of entries, which had no cyclical relationship with the compartments in depolarized light, became significantly symmetrical and bimodal in polarized light with the preferred compartments oriented parallel with thee-vector. Abrupt 90 ° rotation of the vector counter clockwise maintained this relationship during the entire duration of the recordings (up to 17 hours) (Fig. 2). The mean of the orientation angles of the fish on leaving compartments aligned with thee-vector were significantly higher than those from the remaining compartments (Fig. 3). This behavior tended to keep the locomotor orientation parallel with thee-vector as the fish moved between compartments. A strong cyclical relationship between these orientation angles and the compartments of origin was present in polarized but absent in depolarized light. Counter clockwise 90 ° rotation of thee-vector maintained the cyclic behavior of angles but the relationship between the larger means and thee-vector shifted over one or two compartments. This shift disappeared in clockwise rotation. This phenomenon may be due to one of these directions being unnatural. The results demonstrate a pronounced sensitivity and response toe-vector orientation in the goldfish. The sensory mechanism remains unknown.The authors are greatly indebted to Dr. T. H. Waterman for a critical review and discussion of the results here presented.     

Rat osseous plate alkaline phosphatase: mechanism of action of manganese ions

Polidocanol-solubilized osseous plate alkaline phosphatase was modulated by manganese ions in a similar way as by zinc ions. For concentrations up to 1.0 nm, the enzyme was stimulated by manganese ions, showing site-site interactions (n = 2.2). However, larger concentrations (> 0.1 m) were inhibitory. Manganese ions could play the role of zinc ions stimulating the enzyme synergistically in the presence of magnesium ions (K _d = 7.2 m; V = 1005.5 U mg^–1). Manganese ions could also play the role of magnesium ions, stimulating the enzyme synergistically in the presence of zinc ions (K _d = 2.2 m; V = 1036.7 U mg^–1). However, manganese ions could not substitute for zinc and magnesium at the same time since ion assymetry is necessary for full activity of the enzyme. A steady-state kinetic model for the modulation of enzyme activity by manganese ions is proposed.     

Simulation of chaotic EEG patterns with a dynamic model of the olfactory system

The main parts of the central olfactory system are the bulb (OB), anterior nucleus (AON), and prepyriform cortex (PC). Each part consists of a mass of excitatory or inhibitory neurons that is modelled in its noninteractive state by a 2nd order ordinary differential equation (ODE) having a static nonlinearity. The model is called a KO_e or a KO_t set respectively; it is evaluated in the open loop state under deep anesthesia. Interactions in waking states are represented by coupled KO sets, respectivelyKI _e (mutual excitation) andKI _i (mutual inhibition). The coupledKI _e andKI _i sets form aKII set, which suffices to represent the dynamics of theOB, AON, andPC separately. The coupling of these three structures by both excitatory and inhibitory feedback loops forms aKIII set. The solutions to this high-dimensional system ofODEs suffice to simulate the chaotic patterns of the EEG, including the normal low-level background activity, the high-level relatively coherent bursts of oscillation that accompany reception of input to the bulb, and a degenerate state of an epileptic seizure determined by a toroidal chaotic attractor. An example is given of the Ruelle-Takens-Newhouse route to chaos in the olfactory system. Due to the simplicity and generality of the elements of the model and their interconnections, the model can serve as the starting point for other neural systems that generate deterministic chaotic activity.Supported by a grant MH06686 from the National Institute of Mental Health     

Localization of ferruginol,a diterpene phenol,in <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> heartwood by time-of-flight secondary ion mass spectrometry

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) was applied to the investigation of heartwood extractives in Sugi (Cryptomeria japonica). Sugi heartwood tissue generated secondary ions that were not produced from sapwood tissue by TOF-SIMS. Among the peculiar ions generated from heartwood, two positive ions of m/z 285 and 301 were remarkable due to their appearance in a larger mass range and with a high intensity. These two ions were not generated from heartwood tissue preextracted with n-hexane, and the n-hexane extract of Sugi heartwood produced both ions. Gas chromatography-mass spectrometry of the n-hexane extract demonstrated that ferruginol, a diterpene phenol, the molecular weight of which is 286, constituted one of the predominant constituents of the extract. Authentic ferruginol also generated both ions by TOF-SIMS. The molecular formula of the m/z 285 ion generated from Sugi heartwood tissue was estimated to be C₂₀H₂₉O, which corresponds well with that of ferruginol, i.e. C₂₀H₃₀O, by peak identification. All these results strongly suggest that the m/z 285 ion generated from Sugi heartwood tissue originated significantly from ferruginol in Sugi heartwood. By TOF-SIMS imaging, the m/z 285 ion was detected uniformly in the tracheid cell walls, in the cell walls of the axial parenchyma cells and ray parenchyma cells, and also inside these parenchyma cells. These results indicate that ferruginol was distributed almost evenly in Sugi heartwood tissue.     

Lipopeptides Produced by a Soil <Emphasis Type="Italic">Bacillus Megaterium</Emphasis> Strain

A soil microorganism identified as Bacillum megaterium was found to produce several antibiotics substances after growth for 20 h at 37°C in a mineral culture medium. Analysis both by electron spray ionization (ESI) and matrix-assisted laser desorption ionization—time of flight (MALDI-TOF) mass spectrometry (MS) identified these substances as lipopeptides. Predominant peaks at m/z 1,041 and m/z 1,065 revealed ions which are compatible with surfactins and lichenysins, respectively. Two other ions m/z 1,057 and m/z 1,464 were further studied by collision-induced dissociation (CID) unveiling an iturin A at the first and fengycins A and B at the second m/z peaks. The CID spectrum of the m/z 1,464 ion also suggests the existence of fengycins A and B variants in which Ile was changed to Val in the position 10 of the peptide moiety. Raw mixtures of all these compounds were also assayed for antibiotic features. The data enlighten the unusual diversity of the lipopeptide mixture produced by a sole Bacillus species.     

Balancing robust quantification and identification for iTRAQ: Application of UHR‐ToF MS

iTRAQ reagents allow the simultaneous multiplex identification and quantification of a large number of proteins. Success depends on effective peptide fragmentation in order to generate both peptide sequence ions (higher mass region, 150–2200 m/z) and reporter ions (low mass region, 113–121 m/z) for protein identification and relative quantification, respectively. After collision‐induced dissociation, the key requirements to achieve a good balance between the high and low m/z ions are effective ion transmission and detection across the MS/MS mass range, since the ion transmission of the higher m/z range competes with that of the low m/z range. This study describes an analytical strategy for the implementation of iTRAQ on maXis UHR‐Qq‐ToF instruments, and discusses the impact of adjusting the MS/MS ion transmission parameters on the quality of the overall data sets. A technical discussion highlights a number of maXis‐specific parameters, their impact of quantification and identification, and their cross‐interactions.