Novel occurrence of 5 beta-cholanic acid in plants: isolation from jequirity bean seeds (Abrus precatorius L.)

The presence of 5β-cholanic acid has been detected in the seeds of jequirity bean ($\text{Abrus}$$\text{precatorius}$ L.) Its structure was unequivocally established by spectroscopic methods. This report provides proof, for the first time, of the occurrence of 5β-cholanic acid in a plant source.     

Leukotriene A: stereochemistry and enzymatic conversion to leukotriene B

Leukotriene A was assigned the structure 5(S)-$\text{trans}$-5,6-oxido-7,9-$\text{trans}\text{-11,14-}\text{cis}$-eicosatetraenoic acid by the enzymatic conversion of a synthetic product of known stereochemistry into the naturally occurring isomer of 5(S),12(R)-dihydroxy-6,8,10,14-eicosatetraenoic acid in human polymorphonuclear leukocytes.     

The effect of high levels of molasses in combinations with hay on digestibility of organic matter,microbial protein synthesis and volatile fatty acid production in vitro

Fermentation in vitro of $\text{0}\text{100}$, $\text{10}\text{90}$, $\text{40}\text{60}$ and $\text{70}\text{30}$ cane molasses/hay mixtures in media containing urea, trace minerals and vitamins showed a progressive improvement of digestible organic matter in vitro (INVDOM) of hay as the level of molasses increased. Replacement of urea by soya bean meal induced a lower INVDOM of hay in the mixture with 70% molasses.Structural carbohydrates (hay) induced higher microbial synthesis per unit digestible organic matter (DOM) than simple sugars (molasses).The results observed emphasized the importance of adding urea to molasses for improved digestion of hay and a higher percentagge of propionic acid in the rumen. The effect of formulation of molasses/hay rations with soya bean meal or urea on the pattern of fermentation is discussed.     

11-Trans leukotriene C: A naturally occurring slow reacting substance

A slow reacting substance, produced by murine mastocytoma cells, has been shown to have the structure 5(S)-hydroxy-6(R)-$\text{S}$-glutathionyl-7,9,11-$\text{trans}\text{-14-}\text{cis}\text{-eicosatetraenoic acid (11-}\text{trans}$ leukotriene C, previously referred to as leukotriene C-2) by ultraviolet spectroscopy, amino acid analyses, lipoxygenase conversion and comparisions with a synthetic compound of known structure and stereochemistry.     

Steric analysis of glycerophospholipids by circular dichroism. Stereospecifity of phospholipase D catalyzed transesterification.

The circular dichroism spectra of natural glycerophospholipids and synthetic 1-$\text{sn}$-phosphatidic acid were recorded. 3-$\text{sn}$-phosphatidic acid derivatives were found to show a positive Cotton effect, while 1-$\text{sn}$-phosphatidic acid revealed a negative Cotton effect. The results are interpreted in terms of the carboxyl sector rule. By this method phospholipase D was shown to produce stereospecifically 3-$\text{sn}$-phosphatidyl-1-$\text{sn}$-glycerol when incubated with egg yolk lecithin and exess of glycerol.     

Occurrence of a new polyamine,canavalmine, in the sword bean Canavalia gladiata

A new tetraamine was detected in the seed of sword bean $\text{Canavalia gladiata}$ and named canavalmine. The chemical structure was determined to be NH₂ (CH₂) ₄NH(CH₂) ₃NH(CH₂) ₄NH₂ (1,13-diamino-5,9-diazatridecane) based on gas chromatography-mass spectrometry after derivatization of polyamines with pentafluoropropionic anhydride. The proof of identity was established by comparison of infrared and ¹H-NMR spectra of the tetraamine isolated from sword bean with those of a synthetic compound.     

Absorbance and fluorescence properties of protochlorophyllide in etiolated bean leaves

Primary leaves of 7-to-9 day-old etiolated bean seedlings contain a species of protochlorophyllide which is not transformed to chlorophyllide by light; this pigment species exhibits an absorption peak at 631nm $\text{in}\text{vivo}$ at ?196° and a fluorescence emission peak at 639nm $\text{in}\text{vivo}$ at room temperature. Heat-treatment of etiolated leaves converts the phototransformable protochlorophyllide holochrome to a pigment species with $\text{in}\text{vivo}$ absorption and fluorescence peaks identical to those of endogenous nontransformable protochlorophyllide. Administration of δ-amino-levulinic acid to etiolated leaves causes the synthesis of non-transformable protochlorophyllide with an absorption peak also at 631nm $\text{in}\text{vivo}$ at ?196° but with a fluorescence emission peak at 643nm $\text{in}\text{vivo}$ at room temperature. Heat-treatment of such leaves does not affect the position of these bands. The results indicate that protochlorophyllide which is derived from exogenous δ-amino-levulinic acid is in a physically different state from other forms of protochlorophyllide in the leaf.     

The dye absorption test as an indicator of the nutritive quality of heat-processed navy bean

Under standardized conditions, the absorption of cresol red dye by navy bean meal increased from approximately 2.0 mg g^?1 for raw samples up to 3.8 mg g^?1 for optimally cooked samples. Within this range, a pooled correlation coefficient of 0.93 was obtained between dye binding value and $\text{gain}\text{feed}$ ratio of rats fed on diets comprising, in total, 51 differently processed navy bean meals in five bio-assay experiments. Over-heating of navy bean, as judged by a deterioration in the $\text{gain}\text{feed}$ ratio of rats, caused no further change in dye binding value until samples charred, whereupon dye binding values of the order of 1.5 mg g^?1 were observed. The practicality of the test for indicating the nutritive quality of heat-processed navy bean is discussed in the light of these findings.     

Immobilization of Escherichia coli cells having aspartase activity with carrageenan and locust bean gum

Immobilization of Escherichia coli cells having aspartase activity was carried out by к-carrageenan, or by к-carrageenan and locust bean gum. To enhance operational stability, immobilized cells were treated with a hardening agent, such as glutaraldehyde or glutaraldehyde and hexamethylenediamine. Very active and stable immobilized preparations were obtained when E. coli cells immobilized with к-carrageenan were treated with 85 mm-glutaraldehyde and 85 mm-hexamethylenediamine. The productivities of E. coli cells immobilized with polyacrylamide, к-carrageenan, and к-carrageenan and locust bean gum were compared for production of $\text{l}\text{-}\text{aspartic}$ acid. Among these preparations, E. coli cells immobilized with к-carrageenan and treated with glutaraldehyde and hexamethylenediamine showed the highest productivity.     

Stereochemistry of leukotriene C-1

Leukotriene C-1, a “Slow Reacting Substance” (SRS), has been shown to possess the molecular Structure depicted by V (5(S)-hydroxy-6(R)-$\text{S}$-glutathionyl-7,9-$\text{trans}$-11,14-$\text{cis}$-eicosatetraenoic acid) by its identity with a totally synthetic product of known structure and stereochemistry.     

Enzymatic formation of 14,15-leukotriene A and C(14)-sulfur-linked peptides

Human leukocytes converted [³H]-(S)-15-HPETE into [³H]-14,15-LTA. Rat basophilic leukemia cells transformed 14,15-LTA into two bioactive C(14)-S-linked peptides, which have been characterized as 15(S)-hydroxy-14(R)-S-glutathionyl-5,8$\text{Z}$,10,12$\text{E}$-icosatetraenoic acid and 15-(S)-hydroxy-14(R)-S-cysteinylglycyl-5,8$\text{Z}$,10,12$\text{E}$-icosatetraenoic acid by comparison with synthetic specimens.     

Leukotriene A4: Enzymatic conversion to leukotriene C4

An unstable epoxide, leukotriene A₄ (5(S)-$\text{trans}$-5,6-oxido-7,9-$\text{trans}$-11,14-$\text{cis}$-eicosatetraenoic acid), was earlier proposed to be an intermediate in the conversion of arachidonic acid into the slow reacting substance (SRS), leukotriene C₄. In the present work synthetic leukotriene A₄ was incubated with human leukocytes or murine mastocytoma cells. A lipoxygenase inhibitor, BW755C, was added in order to prevent leukotriene formation from endogenous substrate. Leukotriene C₄ and 11-$\text{trans}$-leukotriene C₄ were the main products with SRS activity. It was not established whether the 11-$\text{trans}$-compound was formed by isomerization at the leukotriene A₄ or C₄ stage.     

A rapid isotope dilution method for analysis of indole-3-acetic acid and indoleacetyl aspartic acid from small amounts of plant tissue

A method is described for analysis of indole-3-acetic acid and indoleacetyl aspartic acid in plant tissue. Methanolic extracts are purified on two small columns prior to HPLC using an electrochemical detector. $\text{In}$,$\text{vivo}$ amounts are calculated by isotope-dilution analysis. The method requires only 1 g of tissue, results in relative high recoveries, and is sensitive to the 20 pmol range or below. Typical results using light-grown $\text{Pisum}$,$\text{sativum}$ cv. Little Marvel tissue are given. This report is the first determination of the natural level of the aspartate derivative in growing plant tissues.     

Gibberellin biosynthesis in a cell-free system from immature seeds of Pisum sativum

A cell-free system from immature pea seeds converts ¹⁴C-labelled $\text{ent}$-kaurene to $\text{ent}$-kaurenol, $\text{ent}$-kaurenal, $\text{ent}$-kaurenoic acid, $\text{ent}$-7α-hydroxykaurenoic acid, and gibberellin A₁₂-aldehyde. The latter becomes converted further to 13-hydroxygibberellin A₁₂, gibberellin A44, gibberellin A₁₂-alcohol, and several unidentified products. Thus the biosynthesis of gibberellins $\text{via ent}$-kaurene is now established for a member of the $\text{Leguminosae}$. It is the first time that 13-hydroxylation of gibberellins has been observed in a cell-free system and that gibberellin A₁₂-alcohol has been obtained in any biological system.     

Coenzyme A: fatty acid synthetase apoenzyme 4'-phosphopantetheine transferase in yeast

A mixture of two pantetheine-free mutant fatty acid synthetases was dissociated and recombined $\text{in}$$\text{vitro}$ to form a hybrid apoenzyme complex. $\text{In}$$\text{vivo}$ the corresponding $\text{Saccharomyces}$$\text{cerevisiae}$$\text{fas}$-mutants exhibit interallelic complementation when crossed with each other and the enzyme synthesized in the resulting diploid contains pantetheine and exhibits overall fatty acid synthetase activity. Accordingly, the hybrid apoenzyme formed $\text{in}$$\text{vitro}$ could be activated to holo-fatty acid synthetase when incubated with coenzyme A and a partially purified yeast cell extract. The enzyme coenzyme A: fatty acid synthetase apoenzyme 4′-phosphopantetheine transferase has thus been identified in yeast. Further studies on the mechanism of fatty acid synthetase holoenzyme formation will now be possible.     

Metabolism of mevalonic acid to long chain fatty alcohols in an insect

A mixture of long chain fatty alcohols (LCFA) with chain lengths of 24 to 30 carbon atoms, as free and esterified components, have been isolated from the aphid, $\text{Schizaphis}\text{graminum}$. Both radiolabelled mevalonic acid and palmitic acid served as effective precursors in the synthesis of LCFA. This is the first time that the $\text{trans}$-methylglutaconate shunt has been shown to be operational for fatty alcohol synthesis and to occur in a lower-evolved animal.     

Inhibition of oligopeptide transport in S. cerevisiae by a peptide-poly (ethylene glycol) conjugate

A soluble macromolecule-peptide conjugate, [(Met)₃-OPEG] inhibited the uptake of Met-Met-[${}^{14}\text{C}$] Met into $\text{S.}\text{cerevisiae}$. Uptake of leucine into this strain was not affected by Met₃-OPEG under identical conditions. Inhibition by the macromolecular inhibitor was competitive (K_I = 5.1 × 10^?5M)and followed the structural requirments of the peptide transport systems in $\text{S.}\text{cerevisiae}$ and $\text{C.}$$\text{albicans}$. These findings give the first example of inhibition of metabolite transport by a synthetic macromolecular competitor.     

The influence of charge on phosphatidic acid bilayer membranes

A complete titration of phosphatidic acid bilayer membranes was possible for the first time by the introduction of a new anaologue, $\text{1,2-}\text{dihexadecyl}\text{-sn-}\text{glycerol-3-phosphoric}$ acid, which has the advantage of a high chemical stability at extreme pH values. The synthesis of this phosphatidic acid is described and the phase transition behaviour in aqueous dispersions is compared with that of three ester phosphatidic acids; $\text{1,2-}\text{dimyristoyl}\text{-sn-}\text{glycerol-3-phosphoric}$ acid, 1,3-dimyristoylglycerol-2-phosphoric acid and $\text{1,2-}\text{dipalmitoyl}\text{-sn-}\text{glycerol-3-phosphoric}$ acid.The phase transition temperatures ($\text{T}{}_{\mathrm{<mtext>t</mtext>}}$) of aqueous phosphatidic acid dispersions at different degrees of dissociation were measured using fluorescence spectroscopy and 90° light scattering. The $\text{T}{}_{\mathrm{<mtext>t</mtext>}}$ values are comparable to the melting points of the solid phosphatidic acids in the fully protonated states, but large differences exist for the charged states.The $\text{T}{}_{\mathrm{<mtext>t</mtext>}}$ vs. pH diagrams of the four phosphatidic acids are quite similar and of a characteristic shape. Increasing ionisation results in a maximum value for the transition temperatures at pH 3.5 ($\text{p}\text{K}{}_1$). The regions between the first and the second $\text{p}\text{K}$ of the phosphatidic acids are characterised by only small variations in the transition temperatures (extended plateau) in spite of the large changes occurring in the surface charge of the membranes. The slope of the plateau is very shallow with increasing ionisation. A further decrease in the H⁺ concentration results in an abrupt change of the transition temperature. The slope of the $\text{T}{}_{\mathrm{<mtext>t</mtext>}}$ vs. pH diagram beyond $\text{p}\text{K}{}_2$ becomes very steep. This is the     

The acid-base properties and kinetics of dissolution of the Fe4S4 cores of Chromatin ferredoxin and high potential iron protein.

From a study of the translation of synthetic polynucleotides in the $\text{E. coli}$ and $\text{B. thuringiensis}$ cell-free systems, it is shown that the stimulation of polypeptide synthesis by spermidine depends on the uracil content of messenger ribonucleic acid. This stimulation can not be fulfilled by any amount of Mg²⁺ in the absence of polyamines.     

Formation of the 11,12-diol as a metabolite of benzo(a)pyrene by rat skin in vivo

The dihydrodiols present as metabolites in rat skin after topical application of ³H-labelled benzo(a)pyrene included a significant amount of radioactivity that cochromatographed with synthetic $\text{trans}$-11,12-dihydro-11,12-dihydroxybenzo(a)pyrene. Treatment of the radioactive metabolite with hot mineral acid gave a product that had chromatographic properties identical to those of the phenol similarly formed from the synthetic dihydrodiol and acetylation of the metabolite yielded a product that cochromatographed with the diacetate of the synthetic dihydrodiol. These observations show that the 11,12-dihydrodiol is formed as a metabolite of BP in rat skin $\text{in vivo}$. The metabolite was not detected in mouse skin.