Electrolyte distribution in rabbit superior cervical ganglion

Abstract— Superior cervical ganglia of the rabbit were removed and analysed for Na⁺, K⁺, Ca²⁺, Mg²⁺ and Cl^?. The mean electrolyte content in μmole/g wet wt. was as follows: Na⁺, 64.7 ± 1.3; K⁺, 65.1 ± 2.7; Ca²⁺, 3.71 ± 0.28; Mg²⁺, 3.70 ± 0.50; and Cl^?, 50.15 ± 2.26. Water content was 0.76 ± 0.01 ml/g wet wt. Extracellular space was 0.37 ± 0.01 ml/g, and the vascular space 0.0238 ± 0.0002. The mean resting potential of the rabbit superior cervical ganglion was – 68.6 mv. After correction for extracellular electrolyte content, the potential differences, E_Na, E_K and E_cl, were estimated to be +33.6 mv, –96.9 mv and -41.1 mv, respectively, in the ganglia. Permeability coefficients for K⁺, Na⁺, and Cl^? were estimated to be 1:0.06:0.02. Replacement of sodium in physiological saline solution by lithium results in a displacement of 94 per cent of the sodium content of the ganglion and 69 per cent of the potassium after 30 min of equilibration.     

What is the correct value for Na,K-ATPase activity in homogenates of cerebral cortex?

Activity of Na, K-ATPase in homogenates of fresh cerebral cortex of rats was compared with that of cortex frozen under different conditions. Activity yields after rapid in situ freezing of the exposed cerebral cortex were twice, higher (26.1 U) than in homogenates of the fresh cortex (13.3 U). Fresh brain kept on ice for 60 and 300 s and subsequently frozen in liquid nitrogen yielded activities comparable to those of the tissue frozen in situ (24.1 U and 24.9 U for 60 s and 300 s periods, respectively). Inhibition of Na, K-ATPase by 10⁻⁷ M vanadate was significantly stronger (38%) in homogenates of the fresh brain then in those of the cortex frozen in situ (28%). High Na, K-ATPase activity (47.6 U) in suspensions of synaptosomal membranes (SM) prepared from fresh cortical homogenates was only slightly inhibited by 10⁻⁷ M vanadate (12%). Various treatments of homogenates or SM suspensions, like increase of piston rotation speed, repeated freezing and thawing procedure or vigorous shaking did not significantly affect the enzyme activity. Mg-ATPase activity and its sensitivity to vanadate was also modified by tissue treatment but the effect was much less pronounced.     

PROPERTIES OF TYROSINE HYDROXYLASE IN PERIPHERAL NERVES

Approximately 80 per cent of tyrosine hydroxylase activity in bovine mandibular nerve and rabbit sciatic nerve was soluble, and the rest of the activity was particle-bound. The soluble enzyme in bovine mandibular nerve was isolated by ammonium sulphate fractionation (25–35 per cent saturation). The enzyme had a pH optimum at 5·9 in Tris-acetate buffer, and at 6·5 in Tris-HCl or phosphate buffer. The enzyme required a tetrahydropteridine cofactor. K_m values toward various tetrahydropteridines such as l -erythro-tetrahydrobiopterin (a probable natural cofactor), 2-amino-4-hydroxy-6-methyltetrahydropteridine, and 2-amino-4-hydroxy-6,7-dimethyltetrahydropteridine were 2 × 10⁻⁵m , 5 × 10⁻⁵m and 4 × 10⁻⁴m , respectively. The K_m value for tyrosine at 1 × 10⁻³m -2-amino-4-hydroxy-6-methyltetrahydropteridine as a cofactor was 5 × 10⁻⁵m . The enzyme activity was markedly stimulated with Fe²⁺ or catalase, but Fe²⁺ gave higher activity. The activity was inhibited with α, α′-dipyridyl, l -α-methyl-p-tyrosine, and various catecholamines. Among catecholamines, dopamine was the most potent inhibitor. l -5-Hydroxytryptophan was an inhibitor as potent as dopamine. Neither d -5-hydroxytryptophan nor 5-hydroxytryptamine inhibited the enzyme. The inhibition by l -5-hydroxytryptophan was partially competitive with tetrahydrobiopterin at concentrations higher than 9 × 10⁻⁵m , and partially uncompetitive at concentrations lower than 9 × 10⁻⁵m . The addition of heparin or lysolecithin did not affect enzyme activity with tetrahydrobiopterin as cofactor.     

Extracts of the Brown Alga Dictyota dichotoma (Hudson) J.V. Lamouroux Alleviate Salt Stress in Rice (Oryza sativa L.) During Germination

Algal extracts provide a safe regime for enhancing crop productivity under stressful conditions. The present study evaluated the efficiency of aqueous and ethanolic extracts of the brown alga Dictyota dichotoma in alleviation of salt stress on germination of rice seeds. Firstly, seeds were germinated using the aqueous extract of D. dichotoma at concentrations of 0, 5, 10, 20, and 50 g L⁻¹, prepared either at room temperature (RTE) or by boiling (BLE). The % germination of rice increased from 84% in non-treated seeds to 100% when treated with 20 g L⁻¹ BLE, although this treatment caused reduced uniformity of germination. Embryo growth was maximum at 20 g L⁻¹ of both extracts with superiority of BLE over RTE. In the second experiment, the effect of 20 g L⁻¹ aqueous and ethanolic extracts relative to a balanced nutrient supply on germination of seeds treated with 0, 40, 90, and 170 mM NaCl was investigated. Salinity reduced % germination with delayed onset but high uniformity of germination, whereas algal amendments counterbalanced the effect of salinity to a greater extent relative to the nutrient supply. Upon withdrawal of salt stress, seeds promptly recovered, with more efficient recovery of seeds exposed to 170 mM than from 90 mM NaCl. The lower recovery of salt-treated seeds compared with the control seed germination suggests that rice suffered from the toxic ion effect of salinity on embryo rather than from the osmotic effect. Extracts of D. dichotoma can enhance and also alleviate salinity stress on rice seed germination.

     

Evidence of cylindrospermopsin uptake and clearance in fish (Oreochromis niloticus) under laboratory conditions

Cylindrospermopsin (CYN) is a cyanotoxin that has raised serious concerns about public health in many parts of the world. It can bioaccumulate and affect the health of aquatic organisms, but despite this, few studies have been conducted on CYN uptake and clearance in fish. In this paper, the authors evaluate the uptake and clearance of CYN in the muscle tissue and viscera of juvenile tilapia (Oreochromis niloticus) after exposure to aqueous extracts and whole cells of Cylindrospermopsis raciborskii (CYN-producer). CYN blended with commercial fish food, and three experiments were conducted. In the first trial, fish food, and aqueous extracts containing 0.31 μg CYN g⁻¹ of food per day, was administered to tilapia for 15 days. In the second trial, fish were provided food and intact cells (5.4 μg CYN g⁻¹ of food per day) for 15 days. In the last trial, they were provided fish food and aqueous extracts (0.8 μg CYN g⁻¹ of food per day) for 12 days, and for the next 10 days, the animals were fed food without toxic cell extracts (to simulate a clearance period). The concentration of CYN in muscle tissue and viscera was analysed using ELISA. In the case of juvenile tilapia, the presence of CYN was higher in viscera than in muscle tissue, and the toxin remained in the tissues even after 10 days without the addition of contaminated food. The results suggest that tilapia represents a potential source of CYN transfer through the food web, and this shows the need for a continuous monitoring of this compound in organisms that are used for human and animal consumption.     

SODIUM: SOME EFFECTS ON BLUEGREEN ALGAL GROWTH1

The growth of heterocystous bluegreen algae in various concentrations of sodium, was examined in axenic culture as well as in situ studies. Anabaena cylindrica Lemm. with no Na⁺ added, suffered from decreased rates of acetylene reduction, ¹⁴C, assimilation, excretion of organic C as well as lower concentrations of chlorophyll a and particulate organic C compared to cultures supplied with 5, 10, and 50 mg Na⁺·l⁻¹ Sodium deficient algae released, extracellularly a higher percentage of previously fixed C as organic C. No differences in any parameter measured were demonstrable among cultures grown with 5, 10, and 50 mg Na⁺·l⁻¹ High nitrate concentrations (20 mg NO₃⁻·l⁻¹) resulted in decreased rates of acetylene reduction and heterocyst numbers in. Na sufficient, and Na deficient cultures: however, decreased, cellular Na content at high NO₃⁻ levels occurred only in N deficient, cultures. Higher percentages of excreted organic C occurred with increasing NO₃⁻ concentrations in Na deficient cultures. Sodium enrichment of natural bluegreen populations with the addition of 50, 100, and 200 mg Na⁺·l⁻¹ elicited neither a stimulatory nor an inhibitory response in photosynthetic C fixation. In contrast, the addition of small amounts of Na⁺ (5 mg·l⁻) resulted in increased C fixation. However, since the Na. concentration of the lake water, at ca. 5 mg Na⁺·l⁻¹, was sufficient for growth of the bluegreens present, sodium, is not assumed to be limiting under most natural conditions. No increase in in situ acetylene reduction rates occurred with additions of sodium.     

Synergy between <Emphasis Type="Italic">Salvia officinalis</Emphasis> L. and some preservatives

The aim of the present study is to investigate the antibacterial activity of Salvia officinalis L. aqueous extracts and its synergistic action with preservatives sodium nitrite, sodium benzoate and potassium sorbate in vitro against selected food spoiling bacteria. Synergy was assessed by the checkerboard assay method and quantitatively represented by the FIC index. Synergistic action was established for aqueous extract/ sodium benzoate, aqueous extract/ potassium sorbate, aqueous extract/ sodium nitrite combinations. Synergy was detected in relation to: Agrobacterium tumefaciens, Bacillus subtilis and Proteus sp. Synergy was established at plant extract and preservative concentrations corresponding up to 1/8 MIC values.     

Sodium relations in Chenopodiaceae: a comparative approach

Sodium relations of 15 species of Chenopodiaceae were studied in seedlings grown on quartz sand at 10 mol m^?3 of sodium and potassium. Uptake of sodium and potassium into whole plants and shoots was followed over 2 weeks. High alkali ion uptake rates were found in all species. The apparent selectivity of alkali ion uptake showed a continuous variation between species, from nearly perfect sodium exclusion to negligible cation selection. K/Na ratios above 6 were found in the shoots of eight species. For most of these plants above ground sodium concentrations were highest in the hypocotyls. However, in Chenopodium hybridum (shoot K/Na = 10) and C. urbicum (shoot K/Na = 17) above ground sodium concentrations were lowest in hypocotyls and highest in leaves, as in those species accumulating larger amounts of sodium. These differences are discussed with respect to the underlying mechanisms of ion regulation.     

UPTAKE OF [3H-METHYL]CHOLINE BY MICROSOMAL, SYNAPTOSOMAL, MITOCHONDRIAL AND SYNAPTIC VESICLE FRACTIONS OF RAT BRAIN

Abstract— Microsomal, mitochondrial, synaptosomal and synaptic vesicle fractions of rat brain took up [³H-methyl]choline by a similar carrier-mediated transport system. The apparent K_m for the uptake of [³H-methyl]choline in these subcellular fractions was about 5 × 10^?5 M. Choline uptake was also observed in microsomal fractions prepared from liver and skeletal muscle. Virtually identical kinetic properties for [³H-methyl]choline transport were found in the synaptosomal fractions prepared from the whole brain, cerebellum or basal ganglia. Countertransport of [³H-methyl]choline from the synaptosomal fraction was demonstrated against a concentration gradient. HC-3 was a competitive inhibitor of the uptake of [³H-methyl]choline in brain microsomal, synaptosomal and mitochondria] fractions with respective values for K_i of 4.0, 2.1 and 2.3 × 10^?5 M. HC-15 was a competitive inhibitor of the transport of [³H-methyl]choline in the synaptosomal fraction, with a K_i of 1.7 × 10^?4 M. Upon entry into the microsomal fraction, 74 per cent of the radioactivity could be recovered as unaltered choline, 10 per cent as phosphorylcholine, 1.5 per cent as acetylcholine and 2.5 per cent as phospholipid. Choline acetyltransferase (EC 2.3.1.6) was assayed with [¹⁴C]acetylCoA in synaptosomal fractions prepared from basal ganglia and cerebellum, and in the 31,000 g supernatant fraction of a rat brain homogenate. Enzyme activity was 11-fold greater in the synaptosomal fraction from the basal ganglia than in that from the cerebellum. HC-3 did not inhibit choline acetyltransferase and there was no evidence for acetylation of HC-3. Our findings suggest that choline uptake is a ubiquitous property of membranes in the CNS and cannot serve to distinguish cholinergic nerve endings and their synaptic vesicles.     

Potassium Loss during Galvanotaxis of Slime Mold

The posterior reticulated regions of the plasmodia of the slime mold, Physarum polycephalum, whose migration has been oriented by direct current (3.0 to 5.0 µa/mm² in the agar substrate), contain 30 per cent less potassium than the advancing non-reticulated region. The anterior regions have the same potassium concentration as that of the controls, approximately 32 meq/kg wet weight. Differences in potassium concentration between anterior and posterior regions of control plasmodia, not oriented by electric current, are less than 5 per cent. Sodium, in contrast to potassium, is generally less concentrated in the anterior than in the posterior regions of electrically oriented plasmodia, but sodium concentrations are extremely variable. No significant difference in protein concentration was found between oriented and control plasmodia. Thirty-five per cent of the total potassium, but none of the sodium, is found in acidified ethanol precipitates from plasmodial homogenates. Potassium, but not sodium, appears to be closely associated with processes which differentiate anterior from posterior in an oriented plasmodium.     

Inhibition of the spoilage yeast Pichia occidentalis in a wheat germ diet for mass rearing of Drosophila suzukii

A wheat germ diet was initiated for the mass rearing of Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) and subsequent production of its parasitoid Trichopria drosophilae Perkins (Hymenoptera: Diapriidae). However, the main problem of the diet was the microbial contamination affecting the batches of good pupae production. Therefore, the concentration of four antimicrobials was optimized through the Taguchi orthogonal array for the inhibition of microbial contamination, identified as Pichia occidentalis (Kurtzman et al.) Kurtzman et al. The optimized Taguchi-selected-antimicrobials concentrations were 1.33 g l⁻¹ of potassium sorbate, 1.33 g l⁻¹ of propyl paraben, 1.33 g l⁻¹ of sodium propionate, and 0.16 g l⁻¹ of triclosan. When the antimicrobials were added to the diet, colony-forming units of P. occidentalis were inhibited by two orders of magnitude. Such inhibition means that the D. suzukii females produced 61.1 and 79.3% more pupae and adults, respectively, than the diet with no antimicrobials. These results increase the potential of the wheat germ diet as an artificial diet for mass rearing of D. suzukii and T. drosophilae.     

Subcellular distribution of potassium, sodium, magnesium, calcium and chloride in cerebral cortex

The subcellular distributions of potassium, sodium, magnesium, calcium and chloride have been determined for rabbit cerebral cortex. After homogenization and differential centrifugation, the following percentages of ions were associated with the particulate fraction (nuclear, mitochondrial, synaptic vesicles, and microsomal): (a) 19% of the total potassium; (b) 22% of the total sodium; (c) 77% of the total calcium; (d) 69% of the total magnesium; and (e) less than 2% of the total chloride. However, the sum of the potassium and sodium content in each of the particulate fractions was greater than the sum of the calcium and magnesium content. After hypo-osmotic shock of the crude mitochondrial fraction (M_T), more sodium than potassium (μmol/g wet wt.) was associated with the mitochondrial (M₁) and synaptic vesicle (M₂) fractions. The molar ratio of sodium to potassium was 1·4 for M₁ and 4·5 for M₂. The association of ²²Na⁺ with the particulate fractions was further studied by the method of equilibrium dialysis. The data from both types of experiments indicate that a large fraction of the sodium in cortical tissue appears to be in a bound state.     

The effects of handling procedures,breed differences and treatment with lithium and dexamethasone on some blood parameters in sheep

Fifteen aged Merino and 15 aged Border Leicester ewes each divided into 3 groups of 5 for infusion with lithium chloride, lithium chloride plus dexamethasone and normal saline, and then subjected to 3 jugular venous blood samplings, 1 h apart, in a 3 × 3 × 2 experimental design involving times × treatments × breeds.The blood samples were examined for packed cell volumes, plasma and erythrocyte sodium and potassium concentrations, and plasma calcium concentrations.There were significant changes in packed cell volumes (PCV) 39 v. 30%; 39 v. 31%), plasma sodium concentrations (151 v. 149 mmol l⁻¹; 151 v. 148 mmol l⁻¹) and plasma potassium concentrations (5.3 v. 4.6 mmol l⁻¹; 5.3 v. 4.7 mmol l⁻¹) between Times 0 and 1 and between Times 0 and 2, respectively. There were no significant changes in plasma calcium or erythrocyte sodium or potassium concentrations associated with times. The evidence suggests that the times-effects were caused by different methods of handling the sheep immediately prior to each blood sampling, and this is discussed. The fall in PCV was greater than that recorded by other authors.There were highly significant (P < 0.01) breed differences in PCV (36 v. 31%), plasma calcium concentrations (2.0 v. 2.2 mmol l⁻¹) and erythrocyte potassium concentrations (10.1 v. 15.0 mmol l⁻¹) for Merino and Border Leicester ewes, respectively. There were no significant breed differences in plasma potassium or erythrocyte sodium concentrations.The mean plasma potassium concentration of 5.08 mmol l⁻¹ for the lithium-treated sheep was significantly higher than the means of 4.67 and 4.77 mmol l⁻¹ for lithium plus dexamethasone and saline-treated groups, respectively. There was no significant difference between the latter two means, and there were no significant treatment effects for any of the other blood constituents.     

Rapid Sample Processing for Detection of Food-Borne Pathogens via Cross-Flow Microfiltration

This paper reports an approach to enable rapid concentration and recovery of bacterial cells from aqueous chicken homogenates as a preanalytical step of detection. This approach includes biochemical pretreatment and prefiltration of food samples and development of an automated cell concentration instrument based on cross-flow microfiltration. A polysulfone hollow-fiber membrane module having a nominal pore size of 0.2 μm constitutes the core of the cell concentration instrument. The aqueous chicken homogenate samples were circulated within the cross-flow system achieving 500- to 1,000-fold concentration of inoculated Salmonella enterica serovar Enteritidis and naturally occurring microbiota with 70% recovery of viable cells as determined by plate counting and quantitative PCR (qPCR) within 35 to 45 min. These steps enabled 10 CFU/ml microorganisms in chicken homogenates or 10² CFU/g chicken to be quantified. Cleaning and sterilizing the instrument and membrane module by stepwise hydraulic and chemical cleaning (sodium hydroxide and ethanol) enabled reuse of the membrane 15 times before replacement. This approach begins to address the critical need for the food industry for detecting food pathogens within 6 h or less.     

Quantitative histochemistry of nicotinamide adenine nucleotides in retina of monkey and rabbit

The levels of NADP⁺, NADPH, NAD⁺ and NADH were measured in the different layers of retinas from rabbit and monkey. Samples (0.1 μg) were dissected from frozen-dried sections. The sum of oxidized and reduced forms was obtained by analysis of samples diluted several thousand fold in 0.02 n -NaOH at 0°. The reduced forms were measured by analysis of the same alkaline preparation after heating to destroy NADP⁺ and NAD⁺. All assays were made at 1:100,000 tissue dilution by enzymic cycling, which is capable of measuring 10⁻¹⁴ moles of nucleotides. Profiles of nicotinamide adenine nucleotide levels werecomparable in monkey and rabbit. Both total NADP and NAD were lowest in the outer segments of the retina and highest in the inner layers. NADP of the outer layers (1-2b) was oxidized to a high degree. This was particularly striking for layer 2b, which is rich in mitochondria. In the inner layers the fraction of NADPH rose to 0.7 of the total NADP. NAD on the contrary was highly oxidized in all ten layers of the retina. Three aspects of these results seem significant: (1) The profile for NADP was not related to the distribution of any of four major NADP-requiring dehydrogenases or their sum; (2) the ratio of total NADP/NADPH in the mitochondrial layer was much higher than expected from studies with isolated mitochondria; and (3) the amount of total NADP was surprisingly high in non-mitochondrial layers.     

Flow injection chemiluminescence determination of 6‐mercaptopurine based on a new system of potassium permanganate–thioacetamide–sodium hexametaphosphate

A novel chemiluminescence method for the determination of 6‐mercaptopurine was established based on 6‐mercaptopurine inhibition of the chemiluminescence emission of potassium permanganate–thioacetamide–sodium hexametaphosphate system. The peak height was proportional to log 6‐mercaptopurine concentration in the range 7.0 × 10^?10 to 1.0 × 10^?7 g/mL and the detection limit was 1.9 × 10^?11 g/mL (S/N = 3). The relative standard deviation was 1.5% for the determination of 8.0 × 10^?8 g/mL 6‐mercaptopurine (n = 11). The proposed sensor was successfully applied to the analysis of 6‐mercaptopurine in human serum samples. Copyright © 2009 John Wiley & Sons, Ltd.     

PRODUCTIVITY OF PODOSTEMUM CERATOPHYLLUM IN THE NEW RIVER,VIRGINIA

Productivity of Podostemum ceratophyllum, the dominant aquatic macrophyte in the New River, was measured at four sites representing soft- and hardwater reaches of the river. Available dissolved inorganic carbon (DIC) was 4–5 times greater in the hardwater reach. The difference in available DIC was reflected in standing crop and productivity of P. ceratophyllum. Maximum standing crops of P. ceratophyllum at the two hardwater sites (Sites 1 and 2) were 244.8 ± 30.7 g ash-free dry wt (AFDW) m^−-2 and 193.8 ± 18.7 g AFDW m^−-2 compared to 128.5 ± 14.9 g AFDW m^−-2 and 101.3 ± 6.9 g AFDW m^−-2 for the softwater sites (Sites 3 and 4). Productivity, based on differences in standing crops, was: Site 1, 1.08 ± 0.12 g C m^−-2 d^−-1; Site 2, 0.86 ± 0.08 g Cm^−-2d^−-1; Site 3,0.58 ± 0.06 g C m^−-2 d^−-1; Site 4,0.45 ± 0.03 g C m^−-2 d^−-1. Corresponding values for productivity as ¹⁴C uptake were: 2.77 ± 0.44 g C m^−-2 d^−-1; 2.10 ± 0.45 g C m^−-2 d^−-1; 0.34 ± 0.04 g C m^−-2 d^−-1; 0.28 ± 0.03 g C m^−-2 d^−-1. Productivity/biomass (P/B) based on ¹⁴C uptake and standing crop revealed that P. ceratophyllum productivity was inhibited at the softwater sites perhaps due to carbon limitation. Because of its abundance and its high productivity, P. ceratophyllum is hypothesized to contribute significantly to the New River organic matter budget.     

Effect of sodium in the nutrient medium on the incidence of potassium-deficiency symptoms in tomato plants

Summary Tomato plants (Lycopersicon esculentum cv. Amberley Cross) were grown in sand culture and were fed with four concentrations of potassium nitrate in combination with two levels of sodium nitrate. After six weeks the plants were scored for the presence and absence of a symptom of potassium deficiency, namely, marginal chlorosis and/or necrosis in the young, fully-expanded leaves. These leaves were also analysed for K and Na. Marginal chlorosis and/or necrosis occurred in plants given a nutrient solution containing 0.5 meq K/I or less and supplied with either of the sodium nitrate levels. However, the symptoms occurred more frequently in plants receiving the lower level of sodium nitrate. The laminae on plants receiving the lower concentration of sodium nitrate had a 50 per cent incidence of chlorisis and/or necrosis when the tissue potassium content was 0.74 per cent of the dry wt, while those laminae on plants receiving the higher level did not show a 50 per cent incidence until their potassium fell to 0.64 per cent of the dry wt.     

THE CONVULSANT ACTION OF METHYLDITHIOCARBAZINATE: A COMPARISON WITH OTHER SULPHUR-CONTAINING HYDRAZIDES

—The convulsant action of methyldithiocarbazinate (MDTC), thiocarbohydrazide (TCH) and thiosemicarbazide (TSC) has been studied in mice. The relationship between dose and time to convulsions indicated that MDTC has a dual action and is more potent than TSC. Pretreatment of mice with pyridoxal phosphate (0.25 mmol/kg) protected against convulsions and death produced by low doses of MDTC or TCH, and low or high doses of TSC. Pretreatment with pyridoxine hydrochloride (0.25 mmol/kg) protected mice against TSC but not against TCH. It protected against low doses of MDTC (0.12 mmol/kg), but shortened the latency to convulsions after intermediate doses of MDTC (0.37 mmol/kg). Glutamate decarboxylase activity (GAD, EC 4.1.1.15) in whole brain homogenates from mice killed at the onset of seizures, was significantly reduced by all 3 drugs at all doses. This inhibition did not exceed 30% after any dose of TSC or TCH, but was 64% in mice killed 4 min after the injection of MDTC (0.98 mmol/kg). The addition of pyridoxal phosphate to brain homogenates abolished GAD inhibition after MDTC but not after TCH. In vitro brain GAD was 50% inhibited by 10⁻⁴m -MDTC, 18% by 10⁻⁴m -TSC and 8% by 10 ⁻⁴m -TCH. Kinetic studies suggested that at low concentrations MDTC inhibits by competing with pyridoxal phosphate. At the onset of convulsions the cerebral content of pyridoxal phosphate was reduced after low or high doses of TSC (0.27 and 2.2 mmol/kg) and after high doses of MDTC (0.98 mmol/kg). All three drugs (at 10⁻⁵−10⁻⁴m ) inhibited pyridoxal phosphokinase (EC 2.7.1.35) in vitro. Short latency convulsions after MDTC (0.37–0.98 mmol/kg) very probably arise from inhibition of cerebral GAD, due to competition for coenzymic sites and/or unavailability of coenzyme. Long-latency convulsions after MDTC (0.12–0.37 mmol/kg) are comparable to those seen after TSC (0.27–2.2 mmol/kg) and may depend on a mechanism additional to inhibition of GAD.     

Antibiofilm Activity of Invasive Plants against Candida albicans: Focus on Baccharis halimifolia Essential Oil and Its Compounds

The extracts of five invasive plants were investigated for antifungal and antibiofilm activities against Candida albicans, C. glabrata, C. krusei, and C. parapsilosis. The antifungal activity was evaluated using the microdilution assay and the antibiofilm effect by measurement of the metabolic activity. Ethanol and ethanol-water extracts of Reynoutria japonica leaves inhibited 50 % of planktonic cells at 250 μg mL⁻¹ and 15.6 μg mL⁻¹, respectively. Ethanol and ethanol-water extracts of Baccharis halimifolia inhibited >75 % of the mature biofilm of C. albicans at 500 μg mL⁻¹. The essential oil (EO) of B. halimifolia leaves was the most active (50 % inhibition (IC₅₀) at 4 and 74 μg mL⁻¹against the maturation phase and 24 h old-biofilms of C. albicans, respectively). Oxygenated sesquiterpenes were the primary contents in this EO (62.02 %), with β-caryophyllene oxide as the major component (37 %). Aromadendrene oxide-(2), β-caryophyllene oxide, and (±)-β-pinene displayed significant activities against the maturation phase (IC₅₀=9–310 μ mol l⁻¹) and preformed 24 h-biofilm (IC₅₀=38–630 μ mol l⁻¹) of C. albicans with very low cytotoxicity for the first two compounds. C. albicans remained the most susceptible species to this EO and its components. This study highlighted for the first time the antibiofilm potential of B. halimifolia, its EO and some of its components.