Alveld-Producing Saponins

Stabursvik (1959) described the saponin fraction of Narthecium ossifragum as a sarsasapogenin glycoside with the structure arabinosegalactose-xylose-glucose-sarsasapogenin. In a renewed study of the phototoxic lamb disease alveld, in which this saponin has been implicated (Ender 1955), we have looked more closely at the saponin fraction. We find that there are two saponins, one major and one minor. Both have a branched trisaccharide on C-3 of the sapogenin. Galactose is directly attached to C-3 in both saponins. The major saponin has glucose and arabinose attached to galactose, the minor saponin has glucose and xylose. We suggest the names narthecin and xylosin for the spirostanol form of these two saponins. In fresh juice from leaves we find little narthecin, however. Most of the saponin is present in the furostanol form, with glucose on C-26. Enzymatic hydrolysis showed this glucose to be bound as a β-glucoside. From specific rotations in partial hydrolysates we conclude that the saccharide on C-3 is a β-D-glucoside, α-L-araboside, β-D-galactoside.     

A review of assessment and remediation strategies for hat spot sediments

Sediments at the extreme end of the scale of contamination, designated as hot spot sediments, are considered. The characterization of such hot spots, and an approach for the quantitative assessments of the behaviour and fate of pollutants in such sediments are covered. Experiments with sediments containing extreme levels of heavy metals showed release rates of 56 mg m^–2 d^–1 of dissolved zinc and 0.004 mg m^–2 d^–1 of dissolved mercury. When these sediments were resuspended, the dissolved fluxes were increased by a factor of 2.2 and 128 for zinc and mercury, respectively. The biological implications of hot spot sediments are dealt with, since sediments are an important habitat for many organisms. Various alternatives for clean-up operations including dredging and capping are discussed.     

Mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D) in soil inoculated with Pseudomonas cepacia DBO1(pRO101), Alcaligenes eutrophus AEO106(pRO101) and Alcaligenes eutrophus JMP134(pJP4): effects of inoculation level and substrate concentration

Mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D) by two Alcaligenes eutrophus strains and one Pseudomonas cepacia strain containing the 2,4-D degrading plasmids pJP4 or pRO101 (=pJP4::Tn1721) was tested in 50 g (wet wt) samples of non-sterile soil. Mineralization was measured as ¹⁴C-CO₂evolved during degradation of uniformly-ring-labelled ¹⁴C-2,4-D. When the strains were inoculated to a level of approximately 10⁸ CFU/g soil, between 20 and 45% of the added 2,4-D (0.05 ppm, 10 ppm or 500 ppm) was mineralized within 72 h. Mineralization of 0.05 ppm and 10 ppm, 2,4-D by the two A. eutrophus strains was identical and rapid whereas mineralization by P. cepacia DBO1(pRO101) occurred more slowly. In contrast, mineralization of 500 ppm 2,4-D by the two A. eutrophus strains was very slow whereas mineralization by P. cepacia DBO1 was more rapid. Comparison of 2,4-D mineralization at different levels of inoculation with P. cepacia DBO1(pRO101) (6×10⁴, 6×10⁶ and 1×10⁸ CFU/g soil) revealed that the maximum mineralization rate was reached earlier with the high inoculation levels than with the low level. The kinetics of mineralization were evaluated by nonlinear regression analysis using five different models. The linear or the logarithmic form of a three-half-order model were found to be the most appropriate models for describing 2,4-D mineralization in soil. In the cases in which the logarithmic form of the three-half-order model was the most appropriate model we found, in accordance with the assumptions of the model, a significant growth of the inoculated strains.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - CFU colony forming units - PTYG peptone, tryptone, yeast & glucose - DPM disintegrations per minute     

Continuous flow microcalorimetric measurement of heat production in white adipose tissue from obese (ob/ob) mice and their lean littermates

Heat production, free fatty acid and glycerol release from white adipose tissue fat pads from obese (ob/ob) mice and their lean littermates are determined. Heat production was significantly lower in obese mice compared to lean mice when expressed on wet weight basis but not when expressed on DNA basis. Noradrenaline significantly increased the heat production in fat pads from both groups of animals. However, the increase in heat production due to noradrenaline addition in fat pads from lean mice was significantly higher than in fat pads from obese mice. The release of free fatty acids and glycerol before incubation with noradrenaline was similar from fat pads from both groups of animals. Addition of noradrenaline to the fat pads increased the release of free fatty acids and glycerol in both groups of animals, but the increase was significantly larger from fat pads from lean mice. In the absence of noradrenaline the free fatty acid/glycerol ratio (mol/mol) in the effluent was 7.9:1 and 4.8:1 for lean mice and obese mice, respectively. In the presence of noradrenaline the ratio decreased to 3:1 for both groups of animals.     

Oxygen uptake of carp,Cyprinus carpio,swimming in normoxic and hypoxic water

Synopsis Oxygen uptake (V_{o 2}) was measured in carp of approximately 40 cm length swimming at controlled variable oxygen tensions (P_{o 2}). At P_{o 2}> 120 mm Hg V_{o 2} increased with an increase in swimming speed from 5.6 to 11.3 cm · sec^–1. Extrapolation of V_{o 2} to zero activity at P_{o 2} = 140 mm Hg revealed a standard O₂ uptake of 36.7 ml O₂ · kg^–1 · h^–1 at 20° C. At the lowest swimming speed (5.6 cm · s^–1) the oxygen uptake increased when the water P_{o 2} was reduced. A near doubling in V_{o 2} was seen at P_{o 2} = 70 mm Hg compared to 140 mm Hg. At higher swimming speeds in hypoxic water V_{o 2} decreased relative to the values at low swimming speeds. As a result the slope of the lines expressing log V_{o 2} as a function of swimming speed decreased from positive to negative values with decreasing P_{o 2} of the water. pH of blood from the caudal vein drawn before and at termination of swimming at P_{o 2} = 70 mm Hg and 100 mm Hg did not show any decrease in relation to rest values at P_{o 2} = 140 mm Hg. Blood lactate concentration did not increase during swimming at these tensions.     

Microphotometric analysis of NADH-tetrazolium reductase and alpha-glycerophosphate dehydrogenase in human quadriceps muscle.

Synopsis In serial cross-sections of human skeletal muscles stained for either NADH-tetrazolium reductase (NADH-TR) or -glycerophosphate dehydrogenase (-GPD), a linear relation was found between the total content of enzyme in a cell (expressed as the thickness of the section) and the absorbance of the formazan reaction product formed. Little variation (<4.8%) was found in the concentration of formazan (absorbance per unit thickness) when the same cell was measured in serial cross-sections of various thicknesses (2–10 m) along a longitudinal distance of at least 200 m along the cell. The reduction in enzyme activity was found to be negligible after aqueous preincubation. A maximum of 10–12% of the formazan produced in the NADH-TR reaction might be the result of nothing dehydrogenase activity, whereas this unspecific reaction might account for up to 20% of the formazan deposited in the -GPD reactions after 30 min incubation. The diffusion of Nitro BT into the tissue during the incubation period was found to be unhindered. The rates of formazan production decreased with increasing incubation time, especially in the -GPD reaction in both fibre types. The ratio of the mean absorbance of the formazan in type I fibres to that in type II fibres (in the same section) was 1.41 (coefficient of variation, 2.5%) in the NADH-TR reaction and 0.68 (coefficient of variation, 3.8%) in the -GPD reaction. These values were not affected either by variations in the incubation time (5–40 min) or by the thickness of the section (2–8 m). The concentrations of NADH-TR and -GPD seem to be constant along the length of the muscle fibre. The histochemical reactions reported, together with measurements of the thickness of the sections, seem suitable for the microphotometric quantification of the two enzymes in single fibres of human skeletal muscles.     

Host plant discrimination within Cruciferae: Feeding responses of four leaf beetles (Coleoptera: Chrysomelidae) to glucosinolates, cucurbitacins and cardenolides

Feeding responses of four Chrysomelidae to six less acceptable plants and to compounds from them were investigated by means of leaf disc tests. Significant differences were found between responses of different species, and plants containing potent feeding inhibitors were always rejected. Cucurbitacins are potent feeding inhibitors to Phyllotreta nemorum, and this species does not eat Iberis species containing these compounds. Cardenolides are potent feeding inhibitors to P. undulata, P. tetrastigma and Phaedon cochleariae, and these three species do not eat the cardenolide containing Cheiranthus and Erysimum.Six different glucosinolates all proved to be stimulatory when applied to pea leaf discs. Although the glucosinolates differed somewhat in their ability to stimulate feeding, no correlation is found between content of glucosinolates and acceptability of the investigated plants. Application of sinigrin to Iberis and Cheiranthus did not improve their acceptability. The presence of glucosinolates is necessary for feeding to occur, but it is less important which glucosinolates are present.Cardenolides and cucurbitacins are suggested to be a second generation of protective compounds in Cruciferae, glucosinolates being the first.

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Zusammenfassung Der Einfluss einiger sekundärer Pflanzenstoffe aus Cruciferen auf die Futteraufnahme von vier Chrysomeliden, die auf dieser Pflanzenfamilie vorkommen, wurde mittels Blattscheiben-Tests untersucht. Cucurbitacine sind starke Frasshemmstoffe für Phyllotreta nemorum, weniger starke Hemmstoffe für P. undulata und schwache Hemmstoffe für P. tetrastigma und Phaedon cochleariae. Iberis-Arten, die Cucurbitacine enthalten, werden von P. nemorum und P. undulata abgelehnt, von den beiden anderen Arten aber akzeptiert. Cardenolid-Glykoside vom Strophanthidin-Typ sind starke Frasshemmstoffe für P. undulata, P. tetrastigma und Phaedon cochleariae. Diese Arten lehnen Cheiranthus-und Erysimum-Arten, die solche Stoffe enthalten, ab. Die Futteraufnahme von P. nemorum wird von diesen Stoffen nicht beeinflusst; P. nemorum akzeptiert Cheiranthus- und Erysimum-Arten.Futteraufnahme fand bei Abwesenheit von Senfölglukosiden nicht statt. Sechs verschiedene Senfölglukoside waren alle imstande, das Aufnehmen von Erbsen-Blattscheiben zu stimulieren. Gewisse Unterschiede in der stimulierenden Wirkung der einzelnen Glukoside wurden gefunden. Das Vorkommen bestimmter Glukoside und die Akzeptabilität der Pflanzen zeigten aber keine Korrelation. Anwesenheit oder Abwesenheit von Frasshemmstoffen beeinflusst die Akzeptabilität der Pflanzenarten mehr als die Anwesenheit bestimmter Senfölglukoside.Wenn Senfölglukoside als eine erste Generation von Abwehrstoffen in Cruciferen aufgefasst werden, können Cucurbitacine in Iberis und Cardenolid-Glykoside in Cheiranthus und Erysimum als eine zweite betrachtet werden.

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The Danish Natural Science Research Council supported the research.     

Characterisation of the endospermal trypsin inhibitor of barley

A trypsin inhibitor was isolated from grains of two row barley (cv. Proctor). The purified protein was identical with the corresponding inhibitor of a six row barley (cv. Pirkka); both proteins showed, a Pi of 7.4. The N-terminal amino acid was phenylalanine and an arginine residue was involved in the active site. Effects of substrate concentration showed that the inhibition was noncompetitive with a K_i of about 0.9 × 10^?7M. An enzyme-inhibitor complex was demonstrated by disc electrophoresis.