DNA Barcoding of an Assembly of Montane Andean Butterflies (Satyrinae): Geographical Scale and Identification Performance.

DNA barcoding is a technique used primarily for the documentation and identification of biological diversity based on mitochondrial DNA sequences. Butterflies have received particular attention in DNA barcoding studies, although varied performance may be obtained due to different scales of geographic sampling and speciation processes in various groups. The montane Andean Satyrinae constitutes a challenging study group for taxonomy. The group displays high richness, with more of 550 species, and remarkable morphological similarity among taxa, which renders their identification difficult. In the present study, we evaluated the effectiveness of DNA barcodes in the identification of montane Andean satyrines and the effect of increased geographical scale of sampling on identification performance. Mitochondrial sequences were obtained from 104 specimens of 39 species and 16 genera, collected in a forest remnant in the northwest Andes. DNA barcoding has proved to be a useful tool for the identification of the specimens, with a well-defined gap and producing clusters with unambiguous identifications for all the morphospecies in the study area. The expansion of the geographical scale with published data increased genetic distances within species and reduced those among species, but did not generally reduce the success of specimen identification. Only in Forsterinaria rustica (Butler, 1868), a taxon with high intraspecific variation, the barcode gap was lost and low support for monophyly was obtained. Likewise, expanded sampling resulted in a substantial increase in the intraspecific distance in Morpho sulkowskyi (Kollar, 1850); Panyapedaliodes drymaea (Hewitson, 1858); Lymanopoda obsoleta (Westwood, 1851); and Lymanopoda labda Hewitson, 1861; but for these species, the barcode gap was maintained. These divergent lineages are nonetheless worth a detailed study of external and genitalic morphology variation, as well as ecological features, in order to determine the potential existence of cryptic species. Even including these cases, DNA barcoding performance in specimen identification was 100% successful based on monophyly, an unexpected result in such a taxonomically complicated group.

Study of the in vivo and in vitro cardiovascular effects of four new analogues of ketanserin: implication of 5-HT2A and alpha1 adrenergic antagonism in their hypotensive effect.

The in vivo and in vitro cardiovascular effects of the novel 5-HT2A/alpha1/H1 antagonist ketanserin analogues QF 0303B, QF 0307B, QF 0311B, QF 0313B were studied in anaesthetized normotensive rats (ANR) and in isolated rubbed rat aorta (IRRA). In ANR, 0.2 mg x kg(-1) i.v. of each compound produced a rapid, remarkable but short-lasting fall in mean arterial blood pressure (MAP) accompanied by bradycardia. All compounds significantly modified the pressor effects induced by 5-hydroxytryptamine (5-HT) and noradrenaline (NA). In IRRA, the compounds inhibited NA- and 5-HT-induced contractions in a competitive fashion. Furthermore, the analogues displayed lower H1-antagonist activity than ketanserin. Compounds tested showed low 5-HT2B affinity and no activity at muscarinic, nicotinic, or 5-HT3 receptors, nor any marked ability to produce smooth muscle relaxation via calcium entry blockade. There is a significant correlation between hypotension reached and inhibition of the 5-HT-induced pressor responses (but not for NA). A certain degree of correlation was observed between hypotensive effect endurance vs. alpha1-adrenoceptor blockade (but not for serotonin). These results indicate that in this series the brief hypotensive activity in ANR is attributed to a 5-HT1A receptor blockade and the duration of the effect is better attributed to an alpha1 adrenoceptor blockade.

Effects of aqueous extracts of six marine microalgae on smooth muscle contraction in rat duodenum and vas deferens.

We report the effects of aqueous extracts of six marine microalgae on responses to various contractile agents in isolated rat duodenum (acetylcholine, calcium chloride, barium chloride) and vas deferens (noradrenaline). Our results suggest that only extracts of Phaeodactylum tricornutum, Skeletonema costatum, and Chlorella stigmatophora may have pharmacological potential. Extracts of these three species had spasmolytic effects on responses to the three contractile agents in duodenum. In vas deferens, P. tricornutum and C. stigmatophora inhibited the contractions induced by noradrenaline, whilst S. costatum showed a non-dose-dependent potentiation. With the aim of elucidating their mechanisms of action, we are attempting to isolate the active principles present in these extracts.

Effects of extracts of Tetraselmis suecica and Isochrysis galbana on the central nervous system.

In the course of our continuing pharmacological studies of marine microalgae, we look now at Tetraselmis suecica Stewart (Prasinophyceae) and Isochrysis galbana Demart (Haptophyceae), assessing the effects which aqueous extracts of these algae exert upon the central nervous system. To this end, we have tested for effects upon the following: spontaneous motor activity, body temperature, motor coordination, dextroamphetamine-induced hypermotility, exploratory conduct, muscle relaxation, catalepsy, conditioned avoidance response, oxotremorine-induced cholinergic syndrome, and pentylenetetrazole-induced convulsions. Our results show that aqueous extracts of both microalgae display action profiles which prove difficult to categorize by standard pharmacological criteria.

A study of glaucine-induced relaxation of rat aorta.

The vasorelaxant effect of glaucine, the major alkaloid of Platycapnos spicata (L.) Bernh., was studied. At concentrations of 10 microM-0.3 mM in normal Krebs solution it was almost equieffective in relaxing K(+)-induced and noradrenaline-induced tension in rat aortic rings without endothelium, with IC50 values of 160 +/- 16 microM and 90 +/- 14 microM respectively. In experiments in a calcium-free medium, 10 microM glaucine strongly inhibited noradrenaline-induced contractions. Glaucine (0.3 mM) did not affect basal uptake of 45Ca, but induced uptake was reduced to 100% (K+) and 97.7% (noradrenaline) of the basal value. These results suggest that glaucine has an intracellular effect and also acts on the cell membrane by blocking voltage-dependent and receptor-operated calcium channels.

Antiserotoninergic activity of 2-aminoethylbenzocyclanones in rat aorta: structure-activity relationships.

The antiserotoninergic activity at the serotonin receptor subtype 2 (5-HT2) of seven new 2-aminoethylbenzocyclanones was determined with respect to serotonin-induced contractions in rat aorta and compared with that of ketanserine (pA2 = 8.87). Competitive antagonism was observed in six compounds (6.72 < or = pA2 < or = 8.12). Three-dimensional structures and molecular electrostatic potential distributions of ketanserine and 2-aminoethylbenzocyclanones were analyzed. Several molecular features correlated with the rank of antiserotoninergic activity. In the case of the cyclanone fragment, the rank of activity was associated with the degree of planarity of the bicyclic system. The steric and electrostatic effects due to the loss of planarity were analyzed. In the case of the amino moiety, activity was associated with a particular spatial pattern defined by the amino nitrogen, the aromatic system, and molecular electrostatic potential minima generated by the oxygen atom.

Activity on the central nervous system of Himanthalia elongata; Part II.

We report the effects on the central nervous system (CNS) and on analgesic activity of a fraction (F2) obtained from a Himanthalia elongata extract. The fraction was assayed for effects on spontaneous locomotor activity, d-amphetamine-induced hypermotility, motor coordination, muscular relaxation, rectal temperature, sodium pentobarbital-induced hypnosis, and pentylenetetrazole-induced convulsions. Analgesic activity was evaluated using the hot plate test and the Randall-Selitto test (1). The fraction caused significant reductions in spontaneous locomotor activity, hypermotility, rectal temperature, and motor coordination and postponed pentylenetetrazole-induced death, but no effect was noted on muscle relaxation or the duration of sodium pentobarbital-induced sleep. The fraction exhibited central analgesic effects in the hot plate and Randall-Selitto tests.

Effects of Chlorella stigmatophora extract on the central nervous system.

At the level of the central nervous system an aqueous extract of the marine microalga Chlorella stigmatophora (Butcher) (Chlorophyceae) showed anti-dopaminergic-type activity in assays of its effects on spontaneous motor response, rectal temperature, motor coordination, amphetamine-induced hypermotility, exploratory conduct, muscle relaxation, catalepsy, conditioned avoidance responses, oxotremorine-induced cholinergic syndrome, and pentylenetetrazole-induced convulsions.

Effects of Skeletonema costatum extracts on the central nervous system.

We report the effects of aqueous extracts of the microalga Skeletonema costatum Greve (Cleve) (Bacillariophyceae) on spontaneous motor activity, rectal temperature, motor coordination, amphetamine-induced hypermotility, exploratory behaviour, muscle relaxation, catalepsy, conditioned avoidance responses, oxotremorin-induced cholinergic syndrome, and pentylenetetrazole-induced convulsions. The S. costatum extract at dosages of 310 and 620 mg/kg works like an antidopaminergic drug with anticholinergic properties, that does not induce catalepsy and with a notable muscle relaxing activity.

Effects of Phaeodactylum tricornutum and Dunaliella tertiolecta extracts on the central nervous system.

Continuing work on the effects of aqueous extracts of the microalgae Phaeodactylum tricornutum Bohlin (Bacillariophyceae) and Dunaliella tertiolecta Butcher (Chlorophyceae) on the central nervous system, we report their effects on spontaneous motor activity, rectal temperature, exploratory behaviour, muscle relaxation, catalepsy, and conditioned avoidance responses. Both extracts showed activity as a CNS depressant and a potential muscle relaxant, the latter more marked in the case of P. tricornutum.

Effects of Himanthalia elongata on the central nervous system of mice.

The effects of a Himanthalia elongata extract were studied on the CNS of mice. A rich protein solution obtained from the seaweed was assayed for myorelaxant, anticonvulsant and analgesic activity and for its effects on spontaneous locomotor activity, amphetamine-induced hypermotility, exploratory behaviour, barbiturate-induced sleep, and body temperature. Very significant reductions in spontaneous motor activity, hypermotility and exploratory behaviour were found. The extract prolonged barbiturate-induced sleep and postponed pentylenetetrazol-induced death. Weak myorelaxant, hypothermic and analgesic effects were also observed, showing that H. elongata can depress the CNS.

Effects of Phaeodactylum tricornutum and Dunaliella tertiolecta extracts on the central nervous system.

We report the effects of aqueous extracts of the microalgae Dunaliella tertiolecta Butcher (Chlorophyceae) and Phaeodactylum tricornutum Bohlin (Bacillariophyceae) on oxotremorin-induced cholinergic symptoms, amphetamine-induced hypermotility, pentylenetetrazole-induced convulsions, and Rota-rod test performance. Both extracts significantly reduced rectal temperature, spontaneous motor activity, and time on the Rota-rod and increased the number of mice falling off the rod before 180 s, but neither protected against pentylenetetrazole-induced convulsions or against oxotremorin-induced tremor, salivation, and diarrhoea.

Hydralazine-relaxing effect on rat aorta is not mediated through changes in ATPase activity.

Hydralazine, a hypotensive agent, induces relaxation on smooth musculature. Several mechanisms related to membrane processes have been proposed to explain its relaxing action. In the present paper, the effects of hydralazine on ATPase activity in rat aorta have been studied. Hydralazine (10(-4)-5 X 10(-3) M concentration-dependently relaxed the isolated rat aortic arterial strips under norepinephrine-, serotonin- and K+-contractures. 5-Hydroxytryptamine contractures were more sensitive to the effects of hydralazine than noradrenaline- and potassium-contractures. We found that hydralazine does not modify ATPase activity on rat aorta homogenates. On the other hand, ATPase activity of rat aorta homogenates is dependent on divalent cations Ca2+ and Mg2+, and a little Na, K-ATPase activity was found.

Effects of Lythrum salicaria extracts on hyperglycemic rats and mice.

Ether extracts of Lythrum salicaria stems and flowers exhibited significant hypoglycemic activity in rats with glucose- and epinephrine-induced hyperglycemia when the animals were given oral doses equivalent to 10 g/kg of starting material. These extracts were also found to be active in alloxan- and streptozotocin-diabetic rats and alloxan-diabetic mice. While both stem and flower extracts reduced the elevated gamma-glutamyl transpeptidase activity induced by streptozotocin in rats, only the stem extract reduced the elevated lactic dehydrogenase activity also seen. The flower extract clearly accentuated the elevated levels of aspartate aminotransferase induced by streptozotocin.

Effects of Lythrum salicaria in normoglycemic rats.

Several extracts from stem and flower of Lythrum salicaria were evaluated for their hypoglycemic activity in rats. Results indicate that ether extracts from stem or flowers at a dose equivalent to 10 g/kg of crude plant material caused a significant reduction in blood glucose 4 h after oral administration and that this effect was associated with an increase of circulating insulin. The effects of extracts on lipid and carbohydrate metabolism were also studied.