Effect of Two Wheat Cultivars Differing in Hydroxamic Acid Concentration on Detoxification Metabolism in the AphidSitobion avenae

Hydroxamic acids (Hx) are wheat secondary metabolites conferring resistance for cereals against aphids. The activity of five enzymatic systems were evaluated in the aphid Sitobion avenae reared on the high-Hx wheat cultivar Chagual and the low-Hx wheat cultivar Huayún for 10 generations. Enzyme solutions were prepared from aphid homogenates and assayed for mixed function oxidases (including cytochrome P-450 monooxygenases and NADPH cytochrome c reductase), glutathione S-transferases, esterases, and catalase. Specific activities per aphid individual of cytochrome P-450 monooxygenases, NADPH cytochrome c reductase, glutathione S-transferases, and esterases were significantly increased in wheat cultivars relative to oat (only marginal increase of esterases in Chagual). Aphids fed on cv. Huayún showed an overall higher induction of enzymatic systems than those fed on cv. Chagual. Comparison of these results with reported effects of Hx on detoxifying enzymes in other insects, including aphids, support the hypothesis that these enzymatic pathways play an important role in the detoxification of toxic host-plant secondary metabolites.     

Evaluation of Dimboa Analogs as Antifeedants and Antibiotics Towards the Aphid Sitobion avenae in Artificial Diets

A total of 25 compounds including benzoxazinones, benzoxazolinones, and N-glyoxylamide derivatives were tested as antifeedants and antibiotics towards the aphid Sitobion avenae in diet bioassays. The antifeedant and mortality indexes increased with the presence of electron-donating groups in the 7 position of the benzoxazinone moiety, the replacement of the oxygen atom by sulfur in the heterocyclic ring, the presence of a hemiacetal instead of an acetal at C-2 of the benzoxazine moiety (and hence the possibility of ring opening), and the presence of a hydroxyl group at C-4 of the benzoxazine moiety (hydroxamic acid) instead of a hydrogen atom (lactam). The results support earlier hypotheses on the chemical bases for the mode of action of these compounds.     

Hydroxamic acid glucosides in honeydew of aphids feeding on wheat

DIMBOA glucoside (2-O-/gb-D-glucopyranosyl-4-hydroxy-7-meth-oxy-1,4-benzoxazin-3-one), the main hydroxamic acid (Hx) in intact wheat plants, was detected in the honey dew ofRhopalosiphum padi feeding on seedlings of six wheat cultivars that differed in their concentration of Hx, suggesting that the chemical circulates in the phloem. Neither the aglucone (DIMBOA) nor its main breakdown product were found in any of the honeydew samples. Honey dew production by aphids caged on seedlings of the wheat cultivars and DIMBOA glucoside concentrations in the honeydew followed biphasic curves when plotted against Hx concentration, suggesting passive ingestion of the chemical from the phloem at low Hx concentrations and limited ingestion due to feeding deterrency by Hx in mesophyll cells at high Hx concentrations. The presence of plant toxins such as Hx glucosides in the phloem sap, the main ingesta of aphids, and in the mesophyll cells, has major implications for plant defense, through a feeding deterrent effect during stylet penetration, and deterrency (antixenosis) along with antibiosis during feeding.     

Effect of DIMBOA,an aphid resistance factor in wheat,on the aphid predatorEriopis connexa Germar (Coleoptera: Coccinellidae)

DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one), a secondary metabolite found in cereal extracts, confers resistance in wheat to aphids. Its effect on beneficial organisms was tested on larvae of the aphid predatorEriopis connexa Germar. Larvae were fed until pupation on artificial diets to which different concentrations of DIMBOA (2–200g/g diet) were added, as well as on aphids that had been feeding on wheat seedlings with different DIMBOA levels (140–440 g/g fresh tissue). In diets, the effect of DIMBOA was greatest on survival of third-instar larvae and on the duration of the second and fourth instars. When aphids were provided as food, those that had fed on a wheat cultivar with an intermediate DIMBOA level led to a significantly longer larval duration in the predator than did those that fed on either low or high DIMBOA cultivars. Shortest predator development times were obtained with aphid prey that had fed on high DIMBOA seedlings. Higher DIMBOA levels in the plant appear to reduce aphid feeding rates (and rates of DIMBOA ingestion), decreasing aphid survival and minimizing the effect of the toxin on the predator.On leave from Departamento de Entomologia, Universidad Nacional Agraria La Molina, Apartado aéreo 456, Lima, Perú.     

Resistance of cereals to aphids: The interaction between hydroxamic acids and UDP-glucose transferases in the aphidSitobion avenue (Homoptera: Aphididae)

UPD-glucose transferases are found in the cytosolic and microsomal fractions of the grain aphidSitobion avenae F. Gel filtration and SDSPAGE revealed that the microsomal fraction contained several forms of the enzyme. The molecular weights of the three most active fractions might be 68,000, 66,000, and 36,500. There was a negative correlation between the enzymes' activity in extracts of aphids and the concentration of DIMBOAaglucone in the winter wheat variety fed on by the aphid. A strong inhibition of the activity of the UPD-glucose transferases was observedin vitro at a concentration of DIMBOA as low as 0.01 mM. There was a greater activity of the enzymes in aphids fed on seedlings of susceptible than on moderately resistant wheat cultivars. Prolonged feeding on resistant cultivars resulted in a further reduction in the activity of the aphid's enzymes. The significance for cereal aphids of the role of their UDP-glucose tranferases in the detoxification of plant allelochemicals and adaptation to resistant varieties of cereals is discussed.     

Effect of Previous Exposure to Hydroxamic Acids in Probing Behavior of Aphid Sitobion fragariae on Wheat Seedlings

We hypothesized that aphids after previous exposure to hydroxamic acids (Hx), a family of secondary plant compounds deleterious to aphids, are able to reduce their subsequent exposure to them. This hypothesis was tested by evaluating the time to produce salivation into a sieve element (SSE) by the aphid Sitobion fragariae on seedlings of two wheat cultivars of Triticum aestivum differing in their concentration of Hx. The total time to produce a first SSE was significantly longer in the high-Hx cultivar; however, the subsequent, second SSE (first SSE after interruption of probing) in this cultivar was significantly reduced, reaching the level observed in the low-Hx plants. Therefore, a strategy to reduce the exposure to secondary compounds was observed only in the second SSE in high-Hx plants. When the experimental plant was replaced by a new unattacked plant after the first SSE, aphids did not change the behavior described, thus excluding an aphid-induced plant susceptibility. The number of cell punctures and accumulated duration was not affected by previous exposure to Hx, either in low or high Hx cultivars. Total time and pathway time but not cell punctures, seem to be the variables affected by previous exposure to Hx.     

DIMBOA Glucoside, a Wheat Chemical Defense, Affects Host Acceptance and Suitability of Sitobion avenae to the Cereal Aphid Parasitoid Aphidius rhopalosiphi

The influence of hydroxamic acids (Hx), plant secondary metabolites associated with aphid resistance in wheat, on the host acceptance and suitability of the aphid Sitobion avenae to the cereal aphid parasitoid Aphidius rhopalosiphi was evaluated. Aphids showed a reduction in mean relative growth rate and in body size in the wheat cultivar with higher Hx level. Reduction in aphid size was related to a decreased success in avoiding parasitoid oviposition. A minor increase in A. rhopalosiphi developmental time was observed in aphids feeding on the higher Hx cultivar. Experiments with different concentrations of DIMBOA glucoside, the main Hx in wheat, in artificial diets showed an increase in parasitoid developmental time at the highest concentration, with no change in other performance variables. The evidence is discussed in relation to the compatible utilization of host-plant resistance and biological control in integrated pest management.     

Presence of a hydroxamic acid glucoside in wheat phloem sap,and its consequences for performance ofRhopalosiphum padi (L.) (Homoptera: Aphididae)

Phloem sap of wheat seedlings differing in whole leaf hydroxamic acid (Hx) concentrations was collected by cutting stylets of feeding aphids. DIMBOA-glucoside was the only Hx-related product found. Concentration of DIMBOA-glucoside in phloem sap showed a tendency to be negatively correlated with aphid performance.     

Ginsenoside Rd Attenuates Mitochondrial Permeability Transition and Cytochrome c Release in Isolated Spinal Cord Mitochondria: Involvement of Kinase-Mediated Pathways

Ginsenoside Rd (Rd), one of the main active ingredients in Panax ginseng, has multifunctional activity via different mechanisms and neuroprotective effects that are exerted probably via its antioxidant or free radical scavenger action. However, the effects of Rd on spinal cord mitochondrial dysfunction and underlying mechanisms are still obscure. In this study, we sought to investigate the in vitro effects of Rd on mitochondrial integrity and redox balance in isolated spinal cord mitochondria. We verified that Ca²⁺ dissipated the membrane potential, provoked mitochondrial swelling and decreased NAD(P)H matrix content, which were all attenuated by Rd pretreatment in a dose-dependent manner. In contrast, Rd was not able to inhibit Ca²⁺ induced mitochondrial hydrogen peroxide generation. The results of Western blot showed that Rd significantly increased the expression of p-Akt and p-ERK, but had no effects on phosphorylation of PKC and p38. In addition, Rd treatment significantly attenuated Ca²⁺ induced cytochrome c release, which was partly reversed by antagonists of Akt and ERK, but not p-38 inhibitor. The effects of bisindolylmaleimide, a PKC inhibitor, on Rd-induced inhibition of cytochrome c release seem to be at the level of its own detrimental activity on mitochondrial function. Furthermore, we also found that pretreatment with Rd in vivo (10 and 50 mg/kg) protected spinal cord mitochondria against Ca²⁺ induced mitochondrial membrane potential dissipation and cytochrome c release. It is concluded that Rd regulate mitochondrial permeability transition pore formation and cytochrome c release through protein kinases dependent mechanism involving activation of intramitochondrial Akt and ERK pathways.     

Detoxification of cereal plant allelochemicals by aphids: Activity and molecular weights of glutathioneS-transferase in three species of cereal aphids

The activity and molecular weights of glutathioneS-transferase were studied in three species of cereal aphid:Metopolophium dirhodum, Sitobion avenae, andRhopalosiphum padi. The highest level of glutathioneS-transferase activity was recorded in extracts fromM. dirhodum and the lowest fromS. avenae, and extracts of larvae were more active than those from adults. The activity of this enzyme was higher in extracts ofS. avenae andR. padi previously fed on a moderately resistant wheat variety than on a susceptible variety. Gel filtration followed by SDS-PAGE revealed three protein bands in the active fractions. The first had a molecular weight of 28,500 and the second of 27,500 and were present in all three species of aphid. The third protein differed in the three species, having a molecular weight of 26,000 inS. avenae, 25,500 inM. dirhodum, and 24,000 inR. padi.     

Homogeneous Nature of Malaysian Marine Fish Epinephelus fuscoguttatus (Perciformes; Serranidae): Evidence Based on Molecular Markers,Morphology and Fourier Transform Infrared Analysis

Taxonomic confusion exists within the genus Epinephelus due to the lack of morphological specializations and the overwhelming number of species reported in several studies. The homogenous nature of the morphology has created confusion in the Malaysian Marine fish species Epinephelus fuscoguttatus and Epinephelus hexagonatus. In this study, the partial DNA sequence of the 16S gene and mitochondrial nucleotide sequences of two gene regions, Cytochrome Oxidase Subunit I and III were used to investigate the phylogenetic relationship between them. In the phylogenetic trees, E. fuscoguttatus was monophyletic with E. hexagonatus species and morphology examination shows that no significant differences were found in the morphometric features between these two taxa. This suggests that E. fuscoguttatus is not distinguishable from E. hexagonatus species, and that E. fuscoguttatus have been identified to be E. hexagonatus species is likely attributed to differences in environment and ability to camouflage themselves under certain conditions. Interestingly, this finding was also supported by Principal Component Analysis on Attenuated Total Reflectance–Fourier-transform Infrared (ATR-FTIR) data analysis. Molecular, morphological and meristic characteristics were combined with ATR-FTIR analysis used in this study offer new perspectives in fish species identification. To our knowledge, this is the first report of an extensive genetic population study of E. fuscoguttatus in Malaysia and this understanding will play an important role in informing genetic stock-specific strategies for the management and conservation of this highly valued fish.     

Interactions of <Emphasis Type="Italic">Bacillus mojavensis</Emphasis> and <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> with a Benzoxazolinone (BOA) and its Transformation Product,APO

The benzoxazolinones, specifically benzoxazolin-2(3H)-one (BOA), are important transformation products of the benzoxazinones that can serve as allelochemicals providing resistance to maize from pathogenic bacteria, fungi, and insects. However, maize pathogens such as Fusarium verticillioides are capable of detoxifying the benzoxazolinones to 2-aminophenol (AP), which is converted to the less toxic N-(2-hydroxyphenyl) malonamic acid (HPMA) and 2-acetamidophenol (HPAA). As biocontrol strategies that utilize a species of endophytic bacterium, Bacillus mojavensis, are considered efficacious as a control of this Fusarium species, the in vitro transformation and effects of BOA on growth of this bacterium was examined relative to its interaction with strains of F. verticillioides. The results showed that a red pigment was produced and accumulated only on BOA-amended media when wild type and the progeny of genetic crosses of F. verticillioides are cultured in the presence of the bacterium. The pigment was identified as 2-amino-3H-phenoxazin-3-one (APO), which is a stable product. The results indicate that the bacterium interacts with the fungus preventing the usual transformation of AP to the nontoxic HPMA, resulting in the accumulation of higher amounts of APO than when the fungus is cultured alone. APO is highly toxic to F. verticillioides and other organisms. Thus, an enhanced biocontrol is suggested by this in vitro study.     

Chemical and Behavioral Evidence for a Third Pheromone Component in a North American Population of the Black Cutworm Moth, Agrotis ipsilon

The sex pheromone of Agrotis ipsilon had been previously identified as a blend of (Z)-7-dodecenyl acetate (Z7–12:Ac) and (Z)-9-tetradecenyl acetate (Z9–14:Ac). A synthetic blend of Z7–12:Ac and Z9–14:Ac (30 g:10 g) is effective in attracting males in the field. In several countries, addition of (Z)-11-hexadecenyl acetate (Z11–16:Ac) to the previously identified blend increases male captures, but this had not been demonstrated in North America. We found Z11–16:Ac, in addition to Z7–12:Ac and Z9–14:Ac, from pheromone gland extracts of females from North America. The mean ratio of Z7–12:Ac, Z9–14:Ac, and Z11–16:Ac produced by individual females was 70.5:14.2:15.3, respectively. In Kentucky, addition of Z11–16:Ac (60 g) to a two-component blend of Z7–12:Ac and Z9–14:Ac significantly increased the trap capture rate in the field. Traps baited with this three-component blend were 3.3 times (1995) and 4.6 times (1996) more effective in capturing male A. ipsilon than the two-component blend. This improved effectiveness resulted in detection of A. ipsilon in 60% more of the sampling periods in the two years. In the wind tunnel, males flew upwind and contacted a rubber septum loaded with a three-component blend including Z11–16:Ac significantly more frequently than they did to any two-component blend. These results demonstrate that Z11–16:Ac is a pheromone component in this North American population of A. ipsilon.     

Spodoptera exigua Oviposition and Larval Feeding Preferences for Pigweed, Amaranthus hybridus, over Squaring Cotton, Gossypium hirsutum, and a Comparison of Free Amino Acids in Each Host Plant

The beet armyworm, Spodoptera exigua (Hübner), can utilize a number of different host plants for oviposition and larval development, but some host plants are preferred over others. This study, using cage choice tests and olfactometer assays, demonstrates the beet armyworm's preference for pigweed, Amaranthus hybridus L., over cotton, Gossypium hirsutum L. Cage and olfactometer choice assays indicated that olfaction plays an important role in host selection by ovipositing females. First instars exhibited no feeding preference, but the more mobile third instars showed a significant feeding preference for excised pigweed leaves. The higher quantities and more diverse accumulations of free amino acids in pigweed might, in conjunction with other physiochemical and mechanical attractant and deterrent factors in the two-plant species system, play roles in the tendency of the beet armyworm to select pigweed over cotton and in providing a superior array of easily absorbed amino acids as compared to cotton.     

Nest Liquid Resources of Several Cavity Nesting Bees in the Genus Centris and the Identification of a Preservative, Levulinic Acid

Twig-nesting species of bees in the genus Centris including C. bicornuta, C. analis, C. vittata, and C. nitida, found in the dry forest of Guanacaste Province of Costa Rica, provision their nests with pollen and nectar, rather than pollen and oil as reported for other Centris species. The liquid contents of the nests of these four species were found to contain sugars including 66–75% fructose, 25–33% glucose, and a trace of sucrose. The sugar concentration averaged 47.2%, slightly higher than most flower nectars. No tri-, di-, or monoglycerides, the main components of the flower oil of Byrsonima crassifolia, were detected in the nest provisions. Although these four Centris species are also known to collect oil from B. crassifolia, the oil appears to be used for activities other than nest provisioning. The liquid nest contents did have a slight goat-like odor, suggesting the presence of short-chain fatty acids, and were found to contain a small amount (less than 1%) of three fatty acids. Two of these, butanoic and octanoic acid, were found in trace amounts and are responsible for the goat-like odor. A third was identified as levulinic acid, which made up about 99% of the nest fatty acid contents. This fatty acid had little odor, but may be important as a fungicidal agent. Attempts to determine the source of the fatty acids, were not successful.     

Antagonism between jasmonate- and salicylate-mediated induced plant resistance: effects of concentration and timing of elicitation on defense-related proteins,herbivore, and pathogen performance in tomato

The jasmonate (JA) and salicylate (SA) signaling pathways in plants provide resistance to herbivorous insects and pathogens. It is known that these pathways interact, sometimes resulting in antagonism between the pathways. We tested how the timing and concentration of elicitation of each pathway influenced the interaction between the jasmonate and salicylate pathways measured in terms of five biochemical responses and biological resistance to caterpillars and bacteria. The salicylate pathway had a stronger effect on the jasmonate pathway than did the reverse. The negative signal interaction was generated by two distinct paths in the plant. A negative interaction in the biochemical expression of the two pathways was most consistent in the simultaneous elicitation experiments compared to when the elicitors were temporally separated by two days. Herbivore bioassays with Spodoptera exigua also consistently reflected an interaction between the two pathways in the simultaneous elicitation experiments. The negative signal interaction reducing biological resistance to the herbivore was also demonstrated in some temporally separated treatment combinations where attenuation of the biochemical response was not evident. Concentration of the elicitors had an effect on the pathway interaction with consistent biochemical and biological antagonism in the high concentration experiments and inconsistent antagonism in the low concentration experiments. The bacterial pathogen, Pseudomonas syringae pv. tomato (Pst), consistently showed reduced lesion development on plants with SA responses activated and, in some experiments, on JA-elicited plants. Resistance to Pst was not reduced or enhanced in dual-elicited plants. Thus, signal interaction is most consistent when elicitors are applied at the same time or when applied at high doses. Signal interaction affected the herbivore S. exigua, but not the pathogen Pst.     

Effects of Water Deficit Stress, Shade, Weed Competition, and Kaolin Particle Film on Selected Foliar Free Amino Acid Accumulations in Cotton, Gossypium hirsutum (L.)

Leaves of cotton plants, Gossypium hirsutum L., stressed by water deficit, reduced daylight, and weed competition, or treated with a kaolin wettable powder formulation were analyzed for levels of 17 free amino acids (FAAs) using reversed-phase high-performance liquid chromatography. Water deficit stress resulted in heightened free proline levels (49.9-fold, P < 0.001) that were correlated with diffusive resistance (seconds per centimeter). Five other FAAs increased, and the amounts of total free essential (for insect growth and development) amino acids and total FAAs also increased (P 0.05). Cotton grown in 50% shade accumulated significantly more free arginine than control plants. In a small-plot weed competition assay, four FAAs increased and three FAAs decreased in association with weed competition, but because free proline levels were not altered and free arginine levels increased, other stresses aside from water deficit, possibly including shading by tall weeds, appear to have caused the changes. In a small-plot kaolin particle film assay, five FAAs were lower in cotton foliage sprayed weekly with kaolin. Because free proline was unaffected and free arginine was lower, it is possible that kaolin's reflectivity heightened light reception. The responses of free proline and arginine to the treatments used in these assays demonstrate that types and degrees of some stresses to cotton can be characterized by accumulations of certain FAAs. The study also demonstrates how some FAA levels can indicate degrees of cotton stress resulting from weed competition and from kaolin particle film application. Porometry and leaf water potential measurements assisted in corroborating some findings of the study.     

From Terpenoids to Aliphatic Acids: Further Evidence for Late-Instar Switch in Osmeterial Defense as a Characteristic Trait of Swallowtail Butterflies in the Tribe Papilionini

We compared the chemical compositions of the osmeterial secretions of fourth and fifth (last) instars of eight swallowtail species of the tribe Papilionini. Four species (Papilio demoleus, P. polytes, P. paris, and P. macilentus) are Asian Rutaceae-feeding swallowtails. The other four (Chilasa epicydes, C. agestor, P. troilus, and P. glaucus) represent more distant clades within the Papilionini and species with larval hosts in other plant families. We conducted a quantitative analysis for six species, but only qualitative analysis for P. glaucus and C. agestor. In all eight species, regardless of larval host plant, secretions of the fourth instar principally consisted of mono- and sesquiterpene hydrocarbons, whereas those of the fifth instar comprised aliphatic acids and their esters. Consistent with earlier findings, our results suggest that this “heterogeneous” pattern of osmeterial chemistry, not seen in other tribes, may characterize the Papilionini as a whole. Unlike those of most Papilio species, the fourth and fifth instars of Chilasa species resemble each other in body coloration. Thus, the heterogeneous osmeterial pattern is not necessarily associated with color change in papilionid larvae. The major terpenoids identified in fourth instar larval secretions from the six species were α-pinene, sabinene, β-myrcene, limonene, β-phellandrene, (Z)-β-ocimene, (E)-β-ocimene, p-mentha-1,4(8)-diene, β-elemene, β-caryophyllene, (E)-β-farnesene, (3Z,6E)-α-farnesene, (Z)-α-bisabolene, germacrene-A, (E)-α-bisabolene, and germacrene-B. The profiles for individual species differed both qualitatively and quantitatively from one another, and certain species also secreted methyl 3-hydroxy-n-butyrate and oxygenated sesquiterpenes in relatively large proportions. Secretions from fifth instars were composed of varying proportions of isobutyric, 2-methylbutyric, and acetic acids, and methyl and ethyl (minor) esters of both isobutyric and 2-methylbutyric acids. The heterogeneity of osmeterial chemistry in the tribe Papilionini may represent fine-tuning of chemical defense in response to shifting predation pressures as the larvae age and grow.     

Improvement of the growth of Arthrospira (Spirulina) platensis from Toliara (Madagascar): Effect of agitation, salinity and CO2 addition

Arthrospira (Spirulina) platensis Toliara isolated from alkaline and salt lakes in the south-western area of Madagascar is a potential source of proteins that could efficiently fight against food deficiency in developing countries like Madagascar. Up to now, productivity in this country has been low, so a better understanding of the growth conditions of this species is needed to improve its production. Growth experiments were undertaken in bubble columns at laboratory scale. The influence of agitation of the culture, medium salinity (ranging from 13 to 35 g L⁻¹) and CO₂ addition (ranging from 0 to 2%, v/v) on growth and protein content was examined. Because Arthrospira cells are fragile, a bubble column without additional mixing gave the best growth. Arthrospira (Spirulina) platensis showed higher specific growth rate (μ_max) and protein content for lower salinity. Addition of 1% of CO₂ improved the productivity by near 60%. The feasability of semi-continuous culture was demonstrated and optimal culture conditions led to a mean productivity of 0.22 ± 0.03 g L⁻¹ d⁻¹, a mean specific growth rate of 0.015 ± 0.002 h⁻¹ and a protein content of 53 ± 2% of total dry weight.     

Evidence for Inhibitory Perturbations on the Amplitude,Gating, and Hysteresis of A-Type Potassium Current,Produced by Lacosamide,a Functionalized Amino Acid with Anticonvulsant Properties

Lacosamide (Vimpat^®, LCS) is widely known as a functionalized amino acid with promising anti-convulsant properties; however, adverse events during its use have gradually appeared. Despite its inhibitory effect on voltage-gated Na⁺ current (I_Na), the modifications on varying types of ionic currents caused by this drug remain largely unexplored. In pituitary tumor (GH₃) cells, we found that the presence of LCS concentration-dependently decreased the amplitude of A-type K⁺ current (I_K(A)) elicited in response to membrane depolarization. The I_K(A) amplitude in these cells was sensitive to attenuation by the application of 4-aminopyridine, 4-aminopyridine-3-methanol, or capsaicin but not by that of tetraethylammonium chloride. The effective IC₅₀ value required for its reduction in peak or sustained I_K(A) was calculated to be 102 or 42 µM, respectively, while the value of the dissociation constant (K_D) estimated from the slow component in I_K(A) inactivation at varying LCS concentrations was 52 µM. By use of two-step voltage protocol, the presence of this drug resulted in a rightward shift in the steady-state inactivation curve of I_K(A) as well as in a slowing in the recovery time course of the current block; however, no change in the gating charge of the inactivation curve was detected in its presence. Moreover, the LCS addition led to an attenuation in the degree of voltage-dependent hysteresis for I_K(A) elicitation by long-duration triangular ramp voltage commands. Likewise, the I_K(A) identified in mouse mHippoE-14 neurons was also sensitive to block by LCS, coincident with an elevation in the current inactivation rate. Collectively, apart from its canonical action on I_Na inhibition, LCS was effective at altering the amplitude, gating, and hysteresis of I_K(A) in excitable cells. The modulatory actions on I_K(A), caused by LCS, could interfere with the functional activities of electrically excitable cells (e.g., pituitary tumor cells or hippocampal neurons).