Two oligosaccharides from Marsdenia roylei

Phytochemical analysis of dried twigs of Marsdenia roylei (family Asclepiadaceae) has resulted in the isolation of a trisaccharide, maryal, and a diglycoside, rolinose. Their structures were determined as O-beta-D-oleandropyranosyl-(1-->4)-O-beta-D-digitoxopyranosyl++ +-(1-->4)-D- cymaral and ethyl O-beta-D-oleandropyranosyl-(1-->4)-O-3-O-methyl-6-deoxy-beta-D- allopyranoside, respectively, by chemical degradation and spectroscopic methods.     

Fermentation parameters of Peptostreptococcus productus on gaseous substrates (CO,H2/CO2)

Intrinsic growth and substrate uptake parameters were obtained for Peptostreptococcus productus, strain U-1, using carbon monoxide as the limiting substrate. A modified Monod model with substrate inhibition was used for modeling. In addition, a product yield of 0.25 mol acetate/mol CO and a cell yield of 0.034 g cells/g CO were obtained. While CO was found to be the primary substrate, P. productus is able to produce acetate from CO₂ and H₂, although this substrate could not sustain growth. Yeast extract was found to also be a growth substrate. A yield of 0.017 g cell/g yeast extract and a product yield of 0.14 g acetate/g yeast extract were obtained. In the presence of acetate, the maximum specific CO uptake rate was increased by 40% compared to the maximum without acetate present. Cell replication was inhibited at acetate concentrations of 30 g/l. Methionine was found to be an essential nutrient for growth and CO uptake by P. productus. A minimum amount of a complex medium such as yeast extract (0.01%) is, however, required.     

Improved media for normal human muscle satellite cells: Serum-free clonal growth and enhanced growth with low serum

Summary We have developed a serum-free medium for clonal growth of normal human muscle satellite cells (HMSC). It consists of an optimized nutrient medium MCDB 120, plus a serum-free supplement, designated SF, that contains epidermal growth factor (EGF), insulin, dexamethasone, bovine serum albumin, and fetuin. Fibroblast growth factor was needed with dialyzed fetal bovine serum (dFBS) as the only other supplement, but in media containing SF, it was only slightly beneficial, and was omitted from the final medium without significant loss. Clonal growth of HMSC in MCDB 120 plus SF is as good as with 15% serum and 0.5% chicken embryo or bovine pituitary extract. However, growth is further improved by use of a doubly-supplemented (DS) medium containing both SF and 5% dFBS. Clonal growth of HMSC in the DS medium far exceeds that in previous media with any amount of serum, and monolayer growth is at least equal to that in conventional media with higher levels of serum. Cells grown in these media exhibit little differentiation, even when grown to high densities. However, they retain the capacity for extensive fusion and synthesis of increased creatine kinase when transferred to a serum-free differentiation-promoting medium, such as Dulbecco's modified Eagle's medium plus insulin. All experiments were done with clonal cultures of HMSC to insure that observed growth responses were always those of muscle cells. This research was supported by a grant from the Muscular, Dystrophy Association. Editor's statement This article describes the optimization of both the basal nutrient medium and growth factor requirements for human muscle cells in vitro. This system is critical for studies of normal muscle cell and molecular biology, as well as for understanding diseases of muscle such as Duchenne, Muscular Dystrophy.     

Supersaturation of soil solutions with respect to gypsum

The products of activities of calcium and sulphate were calculated for solutions of 75 glasshouse soils. The majority of these products was found to be higher than the solubility product of gypsum, thus indicating that these soil solutions were possibly supersaturated. In another investigation, soil solutions were examined to determine whether such high activity products could be really attributed to supersaturation. By means of ultracentrifuging of solutions of glasshouse soils, it could be established that the solutions were practically free of sulphate-bearing colloidal particles. Some solutions contained calcium-bearing colloidal particles, but the quantities of calcium contained in these particles were too small to substantially influence the calcium activity. Addition of gypsum crystals to soil solutions led to crystallization of so much calcium and sulphate that the products of the activities of calcium and sulphate dropped from values that can be listed as high to values approaching the solubility product of gypsum. The results obtained demonstrate the occurrence of supersaturation of soil solutions with respect to gypsum. It is further postulated that the presence of humic substances in the soil solution is responsible for this supersaturation. The possible occurrence of supersaturation with respect to gypsum in soils other than glasshouse soils is discussed.     

Follicular dynamics in mares treated with an equine pituitary extract

The follicular dynamics of 112 mares treated with an equine pituitary extract were studied. Follicles >10 mm in diameter at day 15 post-ovulation appeared to represent the follicles which were induced with pituitary extract to grow and ovulate. This was shown by the greater number of >10 mm follicles in mares which subsequently had higher ovulation rates and by the subsequent decrease in number of small follicles (<20 mm) which corresponded with the increase in number of large follicles (>/=20 mm). The difference in diameter (mm) between the largest and second largest follicle on day 15 post-ovulation was greater (P<0.05) for extract-treated mares which subsequently had single ovulations than for extract-treated mares which subsequently had multiple ovulations (7.7 +/-1.5 vs 2.8 +/-0.6). The observed ratio of bilateral to unilateral multiple ovulations was not different (P>0.1) from the expected ratio which was calculated on the assumption that side of ovulation occurred independently (59:19 vs 62:16, observed vs expected).     

Philomena oncorhynchi: Effect of hormones on maturation in anadromous sockeye, Oncorhynchus nerka

Adult sockeye salmon, Oncorhynchus nerka (Walbaum), treated with salmon pituitary extract in July survived for up to 47 days and developed nuptial colours in both male and female fish during this period, whereas no such characteristics appeared in untreated control fish. Furthermore, results showed that 20 of 50 adult female Philomena oncorhynchi recovered from the 15 treated fish contained fully developed tailed larvae in the uterus as compared with only one of 44 female P. oncorhynchi from 31 untreated control fish. None of 51 worms from 13 stilboestrol treated adult sockeye showed development in utero beyond an elongated embryo, nor did the host fish develop nuptial colours. P. oncorhynchi and its immature sockeye host (approx. 1 year prior to spawning) remained unaffected by any of three separate hormone treatment experiments using injection of salmon pituitary extract, injection of 17-β-estradiol, and stilboestrol mixed in the food.     

Purification of thymocyte growth peptide (TGP) from sheep thymus. Relationship to FTS/thymulin

Thymocyte growth peptide (TGP) initiates DNA synthesis in immature thymocytes and has previously been characterized as an acidic peptide isolated from calf thymus. We now report the isolation of TGP from sheep thymus and show it to be a nonapeptide with a large N-terminal blocking moiety characterized by high UV absorbance. The amino acid composition is identical to FTS, consisting of 2 Gly, 2 Ser, 2 Glx, 1 Ala, 1 Lys, 1 Asx. In contrast to FTS, TGP is acidic with an apparent isoelectric point of 4.2 and a high UV absorbance at 270–280 nm. Reverse phase chromatography of TGP at an acidic pH results in a change of the molecule and the appearance of two new compounds TGP-A and TGP-B, both with less than 50% of the original TGP activity. Full activity could be restored by the addition of ZnCl₂ to TGP-A. Both TGP-A and B have some amino acid composition and high UV absorbance as native TGP. We propose that TGP consists of a non-peptide moiety bound to the N-terminal of the nonapeptide Glu-Ala-Lys-Ser-Gln-Gly-Gly-Ser-Asn and that the active molecule is stabilized by Zn²⁺.     

Turnover of organic nitrogen in soils and its availability to crops

The root development of barley seedlings grown for one week in an aerated nutrient solution was studied in the presence of dissolved organic matter from an aqueous chestnut leaf litter extract. In particular, the different effects of low and high molecular weight fractions (small molecules: molecular weight <1000; large molecules: >10,000) of the leaf litter extract were examined. In the presence of large molecules root growth was inhibited, an irregular root tip morphology was observed, and Ca and Mg concentrations in the shoots were lower than in control plants. These phytotoxic effects were not caused by the formation of an impermeable layer of large molecules on the root surfaces that lower accessibility for nutrient cations as inferred from voltammetric experiments. A germination assay using spruce seeds, however, indicated allelochemical effects of large molecules, which exhibit a higher aromaticity than the small molecules as indicated by spectroscopic characterisation. In the growth experiments with small molecules, no influence on the root development of barley was evident, but an increase of Ca and Mg in the shoots was detected. During these growth experiments, a large amount of the small molecules, mainly simple phenols and amino acids, disappeared from the nutrient solution. The loss of small molecules was most likely the effect of mineralisation.Abbreviations DOC dissolved organic carbon - DOM dissolved organic matter - LLE leaf litter extract - MW molecular weight - HMDE hanging mercury drop electrode     

The effect of ethylene, octanoic acid and a plant-derived smoke extract on the germination of light-sensitive lettuce seeds

The effects of an aqueous plant-derived smoke extract, octanoic acid and ethylene on germination of light-sensitive Grand Rapids lettuce seeds were investigated. The smoke extract brought about a concentration dependent increase in germination and a complete inhibition of germination at high concentrations. Octanoic acid could not induce germination. Ethylene at concentrations over 5 L L^–1 increased lettuce seed germination, but not to the same degree as smoke. Aqueous smoke in combination with ethylene showed a synergistic effect on germination at suboptimal smoke concentrations. At high smoke concentrations the effect of ethylene was almost completely inhibited. Octanoic acid in combination with ethylene brought about a higher level of germination than with ethylene alone, but only at the highest concentration of octanoic acid tested (1 mM). Standardized hexane and dichloromethane-partitioned smoke extracts and octanoic acid were subjected to TLC separation. The R _f -fractions in the smoke lanes showing activity in the lettuce seed bioassay did not correspond to the R _f -value of octanoic acid. As aqueous smoke can withstand autoclaving and can be separated by TLC and HPLC without loosing activity it is unlikely that the activity of aqueous smoke is linked to ethylene. It thus appears that the active compound in smoke is neither octanoic acid nor ethylene.Abbreviations TLC thin layer chromatography - HPLC High performance liquid chromatography     

Extra-weak chemiluminescence of drugs XV. Method for determining stability of Toki-shakuyaku-san extract granules

The purpose of the study was to evaluate the quality of Toki-shakuyaku-san extract granules (TJ-23) using chemiluminescence (CL). A linear relationship was obtained between the log value of the CL of TJ-23 and the reaction temperature. An excellent correlation (r= 0.999) was found between the slope of this curve (ΔA) and the colour intensity due to the browning reaction occurring at the early stage of the Maillard reaction.