Effect of fusaric acid on prooxidant and antioxidant properties of the potato cell suspension culture

Cell suspension cultures of potato (Solanum tuberosum, cv. Tamasha) were treated with fusaric acid (FA), a nonspecific fungal toxin produced by Fusarium species to study the effects of FA on H₂O₂ generation, lipid peroxidation, and activities of antioxidant enzymes, i.e., superoxide dismutase (SOD), catalase, and ascorbate peroxidase (APX). The toxicity of various FA doses was evaluated from viability of cultured cells of S. tuberosum. The toxic concentration of FA (10⁻³ M) reduced cell viability by 32% after 48-h incubation and induced alkalinization of the medium; the nontoxic concentration of FA (10⁻⁶ M) had no effect on cell viability and pH of the culturing medium. The treatment of cells with FA caused rapid reversible accumulation of H₂O₂ in cells, promoted lipid peroxidation, and elevated the activity of antioxidant enzymes. The toxic FA concentration elevated the intracellular H₂O₂ content by 51–59% and stimulated lipid peroxidation rate by 35–40%. The nontoxic FA concentration raised the H₂O₂ content by 84–91% and enhanced lipid peroxidation rate by 18–24%. The addition of FA induced transient biphasic induction of the antioxidant enzymes; the action of toxic and nontoxic concentrations differed in terms of the response amplitudes and dynamics. The results confirm the well-known toxic impact of high doses of FA on the cultured cells, which is determined by membrane transport disorders. In addition, the results reveal that toxic and nontoxic concentrations of FA are able to induce pro- and antioxidant systems in the cultured cells of S. tuberosum.     

Kinetin-Mediated Prolongation of Viability in Recalcitrant Sal (Shorea robusta Gaertn. f.) Seeds at Low Temperature: Role of Kinetin in Delaying Membrane Deterioration during Desiccation-Induced Injury

The effects of kinetin (6-furfurylaminopurine) on viability during storage of recalcitrant sal (Shorea robusta Gaertn. f.) seeds at low temperature (15°C) were investigated. The freshly mature sal seeds showed an absolute loss of viability within 6–7 dah (days after harvest) when stored at ambient or at 15°C (control). Storage of these seeds at 15°C after kinetin (10 ppm) treatment prolonged the viability period up to 35 days with 20% germination. The kinetin-treated seeds exhibited 100% germination up to 10 days compared with 3 days in controls. Measurements of leachate conductivity, ·O⁻ ₂ and lipid peroxidation registered gradual increases from 0 dah onward to 35 dah with significantly low levels compared with controls. On the other hand, an enormous increase in superoxide dismutase activity was discernible for a longer duration (0–35 dah) in kinetin-treated seeds than in control seeds where it remained for 3 dah. The role of kinetin in prolonging seed viability by reducing the loss of leachates, lipid peroxidation, ·O⁻ ₂, and enhancing of superoxide dismutase is discussed. Received October 7, 1997; accepted January 27, 1998     

Lipopolysaccharides of polymyxin B-resistant mutants of Escherichia coii are extensively substituted by 2-aminoethyl pyrophosphate and contain aminoarabinose in lipid A

Lipopolysaccharides (LPS) of two polymyxin-resistant (pmr) mutants and the corresponding parent strain of Escherichia Coli were chemically analysed for composition and subjected to ³¹P-NMR (nuclear magnetic resonance) for assessment of phosphate substitution. Whereas the saccharide portions, fatty acids, and phosphate contents were similar in wild-type and pmr LPS, the latter contained two- to threefold higher amounts of 2-aminoethanol. The pmr LPS also contained 4-amino-4-deoxy-l -arabinopyranose (l -Ara_p4N), which is normally not a component of E. coli LPS. This aminopentose has been assigned to be linked to the 4′-phosphate of lipid A. Comparative ³¹P-NMR analysis of the de-O-acylated LPS of the wild-type and pmr strains revealed that phosphate groups of the pmr LPS were mainly (71-79%) diphosphate diesters, which accounted for only 20% in the wild-type LPS. Diphosphate monoesters were virtually nonexistent in the pmr LPS, whereas they accounted for 42% of all phosphates in wild-type LPS. In the lipid A of the pmr strains, the 4′-phosphate was to a significant degree (35%) substituted by l -Ara_p4N, whereas in the wild-type LPS the l -Ara_pN was absent. In the pmr lipid A₁ 2-aminoethanol was completely substituting the glycosidic pyrophosphate but not the glycosidic monophosphate, forming a diphosphate diester linkage at this position in 40% of lipid A molecules. In the wild-type LPS the glycosidic position of lipid A carried mostly unsubstituted monophosphate and pyrophosphate. Thus the polymyxin resistance was shown to be associated, along with the esterification of the lipid A 4′-monophosphate by aminoarabinose, with extensive esterification of diphosphates in LPS by 2-aminoethanol.     

Lipid-to-gibberellin metabolic switching in Fusarium moniliforme via the action of sesamol

Sesamol (3,4-methylenedioxyphenol) at 2.5 mM inhibited growth of Fusarium moniliforme by about 40% and lipid accumulation by 35%. Gibberellin (GA₃) accumulation was increased by 20-fold, to 63 mg g^–1 biomass, in the presence of sesamol indicating that the acetyl-CoA destined for fatty acid biosynthesis was now being switched into secondary metabolite (GA₃) accumulation. Synthesis of other metabolites from acetyl-CoA, such as bikaverin and carotenoids, though were not increased in the presence of sesamol. Metabolic switching is therefore feasible by judicious use of selected inhibitors that can thus block primary metabolic routes but which do not affect secondary metabolites.     

Characterization of sulphatide-containing lipoproteins in rat brain

—(1) Water-soluble [³⁵S]sulphatide is found in the 105,000 g supernatant (SN) of rat brain after intraperitoneal injection of Na₂³⁵SO₄. This labelled sulphatide has a density between those of free lipid and free protein. (2) Fractionation of SN by preparative acrylamide gel electrophoresis indicates that the [³⁵S]sulphatide is not distributed among all SN proteins, but is associated with certain specific proteins. One of the isolated [³⁵S]sulphatide-containing proteins appears homogeneous by analytical acrylamide gel electrophoresis at several pH values. (3) Comparison of the turnover of [³⁵S]sulphatide in microsomes, SN, and myelin indicates that these three subcellular fractions behave as distinct metabolic pools, which meet the requirements for a precursor-product relationship between microsomes and SN and between SN and myelin. (4) These results suggest that sulphatide, synthesized in the microsomes, is transported to the myelin membrane as water-soluble sulphatide containing Iipoproteins in SN.     

Lipid formation by Cryptococcus albidus in nitrogen-limited and in carbon-limited chemostat cultures

Summary Cryptococcus albidus var. Albidus CBS 4517 was grown in nitrogen-limited and in carbon-limited chemostat cultures. The effect of growth rate and limiting nutrient on lipid accumulation and fatty acid composition was investigated.The maximum lipid content in the biomass was, in both cultivation systems, observed at the lowest dilution rate (growth rate) tested. At this dilution rate, D=0.31 h^-1, cells from the nitrogen-limited culture contained 41% (w/w) lipid and cells from the carbon-limited culture 37%. These results indicate the ability of C. albidus, unlike other oleaginous yeasts, to accumulate lipid also in carbon-limited chemostats.The yield of lipid from carbon source was about the same at D=0.031 h^-1 in nitrogen-limited (Y _L/S=0.16 g/g) as in carbon-limited (Y _L/S=0.17 g/g) cultures and decreased with increasing growth rates. In the nitrogen-limited culture, the lipid productivity was about constant at low growth rates (0.031–0.056 h^-1) and a slight decrease was observed at D=0.08 h^-1, while the specific lipid productivity, q _L, increased to 27.5 mg/g per hour. In the carbon-limited culture, however, lipid productivity increased with increasing growth rates and reached its maximum value near _max, whereas q _L was about constant at 20 mg/g per hour.The fatty acid composition was influenced by the specific growth rate in nitrogen-limited as well as in carbon-limited cultures, although the changes were more pronounced during carbonlimitation. A decrease in the degree of unsaturation (/mole) was also observed with increasing lipid content in the cells.     

Inhibition of xanthine oxidase — xanthine — iron mediated lipid peroxidation by eugenol in liposomes

The effect of eugenol on xanthine oxidase (XO) xanthine(X)-Fe⁺³-ADP mediated lipid peroxidation was studied in liver microsomal lipid liposomes. Eugenol inhibited the lipid peroxidation in a dose dependent manner as assessed by formation of thiobarbituric acid reactive substances. When tested for its effect on XO activity per se, (by measuring uric acid formation) eugenol inhibited the enzyme to an extent of 85% at 10 µm concentration and hence formation of O₂ also However, the concentration of eugenol required for XO inhibition was more in presence of metal chelators such as EDTA, EGTA and DETAPAC, but not in presence of deferoxamine, ADP and citrate. The antiperoxidative effect of eugenol was about 35 times more and inhibition of XO was about 5 times higher as compared to the effect of allopurinol. Eugenol did not scavenge O₂ generated by phenazine methosulfate and NAD but inhibited propagation of peroxidation catalyzed by Fe²⁺ EDTA and lipid hydroperoxide containing liposomes. Eugenol inhibits XO-X-Fe⁺³ ADP mediated peroxidation by inhibiting the XO activity per se in addition to quenching various radical species. (Mol Cell Biochem 166: 65-71, 1997)     

Effect of temperature on water transport through aquaporins

The mean effective water self-diffusion coefficient in maize root segments under the effect of aquaporin blocker (mercuric chloride, 0.1 mM) was measured using the spin-echo NMR method with pulsed magnetic field gradient within the temperature range from 10 to 35 °C. HgCl₂ caused the reduction in water diffusion by 30 % as compared to the control samples. Temperature dependences of water self-diffusion coefficients showed two linear regions with different values of Q₁₀ and activation energy, E_a. As the temperature reduced from 20 to 10 °C, E_a values calculated from the Arrhenius plots were close to those of bulk water (20 ± 3 kJ mol⁻¹) and slightly changed for the sample pretreated HgCl₂. Within the temperature range from 25 to 35 °C the slope of temperature dependences became steeper and E_a values were 31 ± 3 kJ mol⁻¹ for the control and 40 ± 4 kJ mol⁻¹ for the treated sample. In the vicinity of 20 °C, the temperature dependence of water diffusion via the mercury-sensitive water channels showed extreme value. In the region, the specific area of the mercury-sensitive aquaporins was 0.004 % of the total cell surface area. The data indicate that water transfer via aquaporins is sensitive to temperature, and the contributions of the transmembrane pathways (aquaporins, lipid bilayer) differ in different temperature ranges.     

Permeability of Boric Acid Across Lipid Bilayers and Factors Affecting It

Boron enters plant roots as undissociated boric acid (H₃BO₃). Significant differences in B uptake are frequently observed even when plants are grown under identical conditions. It has been theorized that these differences reflect species differences in permeability coefficient of H₃BO₃ across plasma membrane. The permeability coefficient of boric acid however, has not been experimentally determined across any artificial or plant membrane. In the experiments described here the permeability coefficient of boric acid in liposomes made of phosphatidylcholine was 4.9 × 10⁻⁶ cm sec⁻¹, which is in good agreement with the theoretical value. The permeability coefficient varied from 7 × 10⁻⁶ to 9.5 × 10⁻⁹ cm sec⁻¹ with changes in sterols (cholesterol), the type of phospholipid head group, the length of the fatty acyl chain, and the pH of the medium. In this study we also used Arabidopsis thaliana mutants which differ in lipid composition to study the effect of lipid composition on B uptake. The chs1-1 mutant which has lower proportion of sterols shows 30% higher B uptake compared with the wild type, while the act1-1 mutant which has an increased percentage of longer fatty acids, exhibited 35% lower uptake than the wild type. Lipid composition changes in each of the remaining mutants influenced B uptake to various extents. These data suggest that lipid composition of the plasma membrane can affect total B uptake. Received: 15 October 1999/Revised: 11 February 2000     

Biosynthesis of lipids by Rhodosporidium toruloides ATCC 10788

The limiting amount of nitrogen required to trigger lipid accumulation in the oleaginous yeast Rhodosporidium toruloides ATCC 10788 was studied, batchwise, by subjecting washed mid-exponentially grown cells to nitrogen at levels of 10⁻² M down to 10⁻⁴ M per g l⁻¹ of lean cells (2–5% fat content) in a mineral medium where glucose was present at 35 g l⁻¹. The results showed that lipid accumulation always started sometime after nitrogen reached a level of 3 × 10⁻⁵ M and the specific initial lipid productivity was constant. Furthermore, the cells were subjected to nine combinations of temperature and pH, from (25° C, pH 4.5) to (35° C, pH 7.5) in the mineral medium supplemented with 0.5 g l⁻¹ of yeast extract and 1 g l⁻¹ (NH₄)₂SO₄. As was expected, lipid content in the cells was higher at 25° C, but pH around 6.0–7.5 slightly enhanced the effect of lower temperature. The effect of pH was also noticed to affect the size of changes in the temporal profiles of the oil's fatty acid distribution prior to nitrogen depletion, whereas no significant difference in the fatty acid composition of the oil was shown after exhaustion of nitrogen from the medium for all combinations of temperature and pH.     

Effect of temperature and pH on lipid accumulation by Trichoderma reesei

Summary The fungal micro-organism Trichoderma reesei was grown in batch culture with excess glucose at pH values between 2.7 and 4.5 and temperatures between 25°C and 35°C. A maximum lipid concentration of 16.9% of the cell dry weight was achieved at pH 3.2 and a temperature of 27°C. Lipid concentration was shown to be correlated with a calculated maximum specific growth rate (µ _mc ) and the maximum lipid value occurred at µ _mc = 0.10 h^–1. Fatty acid analysis was carried out and found to change with changing pH and temperature. Palmitic (16:0) acid and unusually high proportions of stearic acid (18:0) were commonly present. A conversion of fatty acids to palmitoleic acid (16:1) occurred following an unidentified nutrient limitation other than nitrogen depletion after 70 h of culture. Offprint requests to: D. E. Brown     

Taxonomic description of Pseudomonas rhizovicinus sp. nov., isolated from the rhizosphere of a desert shrub Haloxylon ammodendron

A Gram-negative aerobic bacterium, strain M30-35 ^T, was isolated from the rhizosphere of Haloxylon ammodendron in Tengger desert, Gansu province, northwest China. Our previous research indicated that strain M30-35 ^T can promote the growth of ryegrass (Lolium perenne L.). In this study, strain M30-35 ^T was subjected to a polyphasic taxonomic study. Phylogenetic analysis of the 16S rRNA gene and two other housekeeping genes (gyrB, rpoD) showed that strain M30-35 ^T is a member of Pseudomonas anguilliseptica group. The average nucleotide identity (ANI) scores for strains KMM 3042 ^T and FR1439^T were 76.5% and 83.7%, respectively, and DNA-DNA hybridization (DDH) were 21.6% and 26.6%, respectively, and the rates were less than the threshold range for species determination. The dominant cellular fatty acids of strain M30-35 ^T were C_16:0 (22.7%), summed feature 3 (C_16:1ω7c and/or C_16:1ω6c; 18.5%), summed feature 8 (C_18:1ω7c and/or C_18:1ω6c; 23.1%). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phospholipid and aminophospholipid and the predominant respiratory quinone was ubiquinone (Q9). On the basis of above data, it can be concluded that strain M30-35 ^T represents a novel species in the genus Pseudomonas, for which the name Pseudomonas rhizovicinus sp. nov. is proposed. The type strain is M30-35 ^T (=?MCCC 1K03247^T?=?KCTC 52664 ^T).

     

Effect of daytime CO2 supplement on productivity and biochemical composition of Scenedesmus armatus under outdoor cultivation

The cultivation of Scenedesmus armatus was carried out under outdoor Thailand climate conditions. The highest actual growth rate occurred at around 9:00 a.m. to 3:00 p.m., with a wide pH range of 6.4 to 11. The supply of CO₂ had slight influence on growth characteristics but did exert some observable effects on nutritional accumulations. Adding CO₂ from 2 to 15% by volume in the aeration (0.2 vvm) caused an increase in lipid and protein from 19.8 to 25.6 and 37.8 to 48.2% w/w, respectively, whereas carbohydrate decreased from 42.5 to 26.2% w/w. Scenedesmus armatus cultivated with 2% CO₂-enriched air provided the highest the average of the average biomass productivity of 91.25 mg L^?1 d^?1, which corresponded to a CO₂ fixation of 165 mg CO₂ L^?1 d^?1 with the average lipid, protein, and carbohydrate productivities of 22.24, 38.34, and 30.67 mg L^?1 d^?1.     

Optimization of growth and fatty acid composition of a unicellular marine picoplankton,Nannochloropsis sp., with enriched carbon sources

A unicellular marine picoplankton, Nannochloropsis sp., was grown under CO₂-enriched photoautotrophic or/and acetate-added mixotrophic conditions. Photoautotrophic conditions with enriched CO₂ of 2800 l CO₂ l^–1 and aeration gave the highest biomass yield (634 mg dry wt l^–1), the highest total lipid content (9% of dry wt), total fatty acids (64 mg g^–1 dry wt), polyunsaturated fatty acids (35% total fatty acids) and eicosapentaenoic acid (EPA, 20:53) (16 mg g^–1 dry wt or 25% of total fatty acids). Mixotrophic cultures gave a greater protein content but less carbohydrates. Adding sodium acetate (2 mM) decreased the amounts of the total fatty acids and EPA. Elevation of CO₂ in photoautotrophic culture thus enhances growth and raises the production of EPA in Nannochloropsis sp.     

Optimization of lipid production in the oleaginous yeastApiotrichum curvatum in wheypermeate

Summary Lipid production of the oleaginous yeastApiotrichum curvatum was studied in wheypermeate to determine optimum operation conditions in this medium. Studies on the influence of the carbon to nitrogen ratio (C/N-ratio) of the growth medium on lipid production in continuous cultures demonstrated that cellular lipid content in wheypermeate remained constant at 22% of the cell dry weight up to a C/N-ratio of about 25. The maximal dilution rate at which all lactose is consumed in wheypermeate with excess nitrogen was found to be 0.073 h^-1. At C/N-ratios higher than 25–30 lipid content gradually increased to nearly 50% at C/N=70 and the maximal obtainable dilution rate decreased to 0.02 h^-1 at C/N=70. From these studies it could be derived that maximal lipid production rates can be obtained at C/N-ratios of 30–35 in wheypermeate. Since the C/N-ratio of wheypermeate normally has a value between 70 and 101, some additional nitrogen is required to optimize the lipid production rate. Lipid production rates ofA. curvatum in wheypermeate were compared in four different culture modes: batch, fed-batch, continuous and partial recycling cultures. Highest lipid production rates were achieved in culture modes with high cell densities. A lipid production rate of nearly 1 g/l/h was reached in a partial recycling culture. It was calculated that by using this cultivation technique lipid production rates of even 2.9 g/l/h may be reached when the supply of oxygen can be optimized.Nomenclature C/N-ratio carbon to nitrogen ratio of the growth medium (g/g) - C/N_crit C/N-ratio at which there is just enough nitrogen to allow all carbon source to be converted to biomass - D dilution rate=volume of incoming medium per unit time/volume of medium in the culture vessel (h^-1) - D_max maximum dilution rate (h^-1) - DW cell dry weight - L lipid yield (g storage lipid/g carbon source) - specific growth rate (h^-1) - _max maximum specific growth rate (h^-1) - Q_L lipid production rate (g/l/h) - Y_i molecular fraction of carbon substrate that is converted to storage carbohydrate (C-mol/C-mol) - Y_ls maximal amount of storage lipid that can be produced per mol carbon source (C-mol/C-mol)     

The effects of maternal dietary treatments with natural PPAR ligands on lipid metabolism in fetuses from control and diabetic rats

Maternal diabetes impairs fetal development and growth. We studied the effects of maternal diets enriched in unsaturated fatty acids capable of activating peroxisome proliferator-activated receptors (PPARs) on the concentrations of 15deoxyΔ^12,14PGJ₂ (15dPGJ₂), lipid mass, and the de novo lipid synthesis in 13.5-day fetuses from control and diabetic rats. Diabetes was induced by neonatal streptozotocin administration (90 mg/kg). Rats were treated with a standard diet supplemented or not with 6% olive oil or 6% safflower oil from days 0.5 to 13.5 of gestation. Fetuses from diabetic rats fed with the standard diet showed reduced 15dPGJ₂ concentrations, whereas maternal treatments with olive and safflower oils increased 15dPGJ₂ concentrations. Fetuses from diabetic rats showed increased concentrations of phospholipids and increased synthesis of triglycerides, phospholipids, cholesterol and free fatty acids. Diabetic rat treatments with olive and safflower oils reduced phospholipids, cholesterol, and free fatty acid concentrations and the de novo lipid synthesis in the fetuses. These effects were different from those observed in fetuses from control rats, and seem not to involve PPARγ activation. In conclusion, olive oil- and safflower oil-supplemented diets provide beneficial effects in maternal diabetes, as they prevent fetal impairments in 15dPGJ₂ concentrations, lipid synthesis and lipid accumulation.     

Effects of photoperiod,salinity and pH on cell growth and lipid content of Pavlova lutheri

The aim of the present work was to study the effects of photoperiod, salinity and pH on growth and lipid content of Pavlova lutheri microalgae for biodiesel production in small-scale and large-scale open-pond tanks. In a 250-mL flask, the cultures grew well under 24 h illumination with maximum specific growth rate, μ _max , of 0.12 day^?1 and lipid content of 35 % as compared to 0.1 day^?1 and 15 % lipid content in the dark. The salinity was optimum for the cell growth at 30–35 ppt, but the lipid content of 34–36 % was higher at 35–40 ppt. Algal growth and lipid accumulation was optimum at pH 8–9. Large-scale cultivation in 5-L and 30-L tanks achieved μ _max of 0.13–0.14 day^?1 as compared to 0.12 day^?1 in small-scale and 300L cultures.     

Biosignatures present in a hydrothermal massive sulfide from the Mid-Atlantic Ridge

Mid‐ocean spreading and accompanying hydrothermal activities result in huge areas with exposure of minerals rich in reduced chemicals – basaltic and peridotitic rocks as well as metal sulfide precipitates – to the oxygenated seawater. Oxidation of Fe and S present in these rocks provides an extensive long‐term source of energy to lithotrophs. Investigation of lipid biomarkers and their carbon isotope ratios from a massive iron sulfide of an inactive sulfide mound or inactive chimney sampled at the western flank of the Turtle‐Pits hydrothermal field (Mid‐Atlantic Ridge, 5°S) revealed a unique lipid distribution. The bacterial fauna appears to be dominated by chemolithotrophs with a distinct lipid composition mainly comprising of iso‐branched fatty acids and nonisoprenoidal dialkyl glycerol diethers partially including the very rare macrocyclic cores with 30–35 carbon atoms (including 13,16‐dimethyloctacosane and 5,13,16‐trimethyloctacosane). The Bacteria are accompanied by most likely hydrogen/CO₂‐dependent methanogenic Archaea (e.g. Methanococcus) as well as other Archaea with a different life style (e.g. Ferroplasma). Alike some of the bacterial lipids the archaeal lipids predominantly consist of macrocyclic diethers including one C₄₀ and one C₄₁ isoprenoid. Structural homologues of the latter are so far only reported from a methanogenic archaeum and a Pleistocene sulfur deposit. Compound‐specific analyses of the stable isotope ratios revealed δ¹³C values for the majority of bacterial and archaeal lipid components of about 0‰ (vs. VPDB), indicative for chemolithoautotrophically fixed carbon which is, for distinct pathways, accompanied by only negligible fractionations. However, the presence of methanogenic Archaea is indicated by ¹³C‐depleted isoprenoidal lipids (δ¹³C ~ –50‰) characteristic for certain CO₂‐reducing methanogens synthesizing lipids via acetyl CoA.     

LIPID CLASS,CAROTENOID, AND TOXIN DYNAMICS OF KARENIA BREVIS (DINOPHYCEAE) DURING DIEL VERTICAL MIGRATION1

The internal lipid, carotenoid, and toxin concentrations of Karenia brevis (C. C. Davis) Gert Hansen and Moestrup are influenced by its ability to use ambient light and nutrients for growth and reproduction. This study investigated changes in K. brevis toxicity, lipid class, and carotenoid concentrations in low‐light, nitrate‐replete (250 μmol quanta · m^?2 · s^?1, 80 μM NO₃); high‐light, nitrate‐replete (960 μmol quanta · m^?2 · s^?1, 80 μM NO₃); and high‐light, nitrate‐reduced (960 μmol quanta · m^?2 · s^?1, <5 μM NO₃) mesocosms. Reverse‐phase HPLC quantified the epoxidation state (EPS) of the xanthophyll‐cycle pigments diadinoxanthin and diatoxanthin, and a Chromarod Iatroscan thin layer chromatography/flame ionization detection (TLC/FID) system quantified changes in lipid class concentrations. EPS did not exceed 0.20 in the low‐light mesocosm, but increased to 0.65 in the high‐light mesocosms. Triacylglycerol and monogalactosyldiacylglycerol (MGDG) were the largest lipid classes consisting of 9.3% to 48.7% and 37.3% to 69.7% of total lipid, respectively. Both lipid classes also experienced the greatest concentration changes in high‐light experiments. K. brevis increased EPS and toxin concentrations while decreasing its lipid concentrations under high light. K. brevis may mobilize its toxins into the surrounding environment by reducing lipid concentrations, such as sterols, limiting competition, or toxins are released because lipids are decreased in high light, reducing any protective mechanism against their own toxins.     

Autoradiography of Serotonin 5-HT1A Receptor-Activated G Proteins in Guinea Pig Brain Sections by Agonist-Stimulated [35S]GTPγS Binding

Abstract: G protein activation mediated by serotonin 5-HT_1A and 5-HT_1B/D receptors in guinea pig brain was investigated by using quantitative autoradiography of agonist-stimulated [³⁵S]GTPγS binding to brain sections. [³⁵S]GTPγS binding was stimulated by the mixed 5-HT_1A/5-HT_1B/D agonist L694247 in brain structures enriched in 5-HT_1A binding sites, i.e., hippocampus (+140 ± 14%), dorsal raphe (+70 ± 8%), lateral septum (+52 ± 12%), cingulate (+36 ± 8%), and entorhinal cortex (+34 ± 5%). L694247 caused little or no stimulation of [³⁵S]GTPγS binding in brain regions with high densities of 5-HT_1B/D binding sites (e.g., substantia nigra, striatum, central gray, and dorsal subiculum). The [³⁵S]GTPγS binding response was antagonized by WAY100635 (10 µM) and methiothepin (10 µM). In contrast, the 5-HT_1B inverse agonist SB224289 (10 µM) did not affect the L694247-mediated [³⁵S]GTPγS binding response, and the mixed 5-HT_1B/D antagonist GR127935 (10 µM) yielded a partial blockade. The distribution pattern of the [³⁵S]GTPγS binding response and the antagonist profile suggest the L694247-mediated response in guinea pig brain to be mediated by 5-HT_1A receptors. In addition to L694247, 8-hydroxy-2-(di-n-propylamino)tetralin, and flesinoxan also stimulated [³⁵S]GTPγS binding; their maximal responses varied between 46 and 52% compared with L694247, irrespective of the brain structure being considered. Sumatriptan, rizatriptan, and zolmitriptan (10 µM) stimulated [³⁵S]GTPγS binding in the hippocampus by 20–50%. Naratriptan, CP122638, and dihydroergotamine stimulated [³⁵S]GTPγS binding to a similar level as L694247 in hippocampus, lateral septum, and dorsal raphe. It appears that under the present experimental conditions, G protein activation through 5-HT_1A but not 5-HT_1B/D receptors can be measured in guinea pig brain sections.