Semipreparative enantioseparation of Tröger base derivatives by HPLC

We describe the enantioseparation of functionalized derivatives of the Tr?ger base by HPLC on commercially available chiral stationary phases. Cellulose-derived Chiralcel OJ and brush-type Whelk O1 are demonstrated to be complementary to each other in their scope. On the basis of the results obtained, the separation of selected compounds was successfully transferred onto semipreparative columns. We believe that the availability of enantiopure functionalized derivatives of the Tr?ger base will stimulate the further use of this interesting molecular scaffold in molecular recognition, asymmetric catalysis, and related areas of research and technology.     

Sex differences in reproductive investment: maternal care reduces escape response capacity in the whelk Buccinum undatum

We evaluated the effect of reproductive state and biochemical status of male and female whelks Buccinum undatum on their escape responses (foot contortions) when touched by their major predator, the sea star Leptasterias polaris. Gonad growth in males was not associated with a decrease in their general energetic and metabolic capacity, nor in their capacity for escape responses, but did reduce the rate of foot contortions during recuperation from exhaustive exercise. Similarly, the energetic status of females was good when the ovary was mature, suggesting that oogenesis did not require somatic reserves. On the other hand, the energetic status and muscle metabolic capacities of females dropped with egg laying. This decrease was associated with a reduced capacity for escape responses. Thus, egg laying resulted in (1) major drops in the carbohydrate and protein contents of the foot, (2) decreased activities of several glycolytic enzymes (LDH, ADH and APK) in the foot muscle, (3) a sharp drop in the digestive gland index, (4) a decrease in the number of foot contortions and (5) a decreased ability to recover from exhaustive escape exercise. Reproductive costs are much greater for females than males (as females must produce protective egg capsules, search for egg-laying sites and lay the capsules). Females have greater escape capacities than males, except directly after egg laying when their energetic reserves are virtually depleted. The greater capacity of females for escape responses may attenuate the risks associated with their greater boldness in stealing food from their predator L. polaris, particularly prior to egg laying.     

Calcium dependency and the effect of calcium antagonists on molluscan proboscis smooth muscles from the whelk,Buccinum undatum

In all four proboscis muscles of the whelk Buccinum undatum, the potassium-induced depolarization response was acutely dependent upon extracellular calcium, being eliminated in calcium-free conditions. The responses to acetylcholine were found to be partly dependent upon intracellular calcium. Responses to the peptides phenylalanine-methionine-arginine-phenylalanine-NH₂ and phenylalanine-leucine-arginine-phenylalanine-NH₂ were much more resistant to calcium-free conditions and appeared to engage the excitation-contraction coupling mechanism by mobilizing stored intracellular calcium. Sucrose-gap studies of radular retractor muscles showed that the organic calcium “antagonist” nifedipine enhanced potassium-induced depolarization responses, initiating spike-like action potentials and associated fast twitch activity. The inorganic calcium antagonist gadolinium exerted concentration-dependent inhibitory actions on these muscles. Basal tonus and fast twitch activity in response to potassium-induced depolarization were eliminated as were the spike-like action potentials of the membrane electrical response. The inorganic calcium “antagonist” cadmium greatly enhanced potassium-induced contractures in all four muscles, and on its own it induced tonic force and fast twitches in all the muscles. It seems likely that cadmium may have displaced stored intracellular calcium to induce myofilament activation. While these molluscan smooth muscles appear to possess calcium channels with fast and slow characteristics, their behaviour and pharmacological manipulation is very different from their more well known mammalian transient and long-lasting channel counterparts.     

Effect of the mobile phase on the retention behavior of optical isomers of the mandelic acid derivative methyl 2-phenyl-2-(tetrahydropyranyloxy) acetate by chiral HPLC

Conditions for separation of enantiomers of a mandelic acid derivative, methyl 2-phenyl-2-(tetrahydropyranyloxy) acetate (the analyte) were studied. Because of the presence of two chiral carbons, the analyte consists of four stereoisomers stable at ambient temperature. Chiral HPLC of the analyte resulted in four peaks, using an (S,S)-Whelk-O1 column with the mobile phase consisting of hexane and the t-butyl methyl ether (TBME). It was found that TBME dramatically changed the retention of the isomers, though it produced the best enantioseparation on (S,S)-Whelk-O1. The amount of TBME in the mobile phase influenced the degree of retention shift; 5% (v/v) TBME gave a bigger shift than 8% (v/v) and 10% (v/v). 2-Propanol did not produce the same results. The chiral separation was also tried on cellulose tris (3, 5-dimethyl phenylcarbamate) (CDMPC), but only three peaks were seen, indicating some but not full enantiomer resolution.     

The relation between excitation-contraction coupling and fine structure of a molluscan muscle,the radular retractor of the whelk,Buccinum undatum

The Buccinum radula is of the rachiglossate type with two outer rows of fierce hook-like attack teeth and a medial row of straight sharp-pointed shredding teeth. Individual cells of the radular retractor muscle are 10–12 m in diameter and separated at the closest by gaps of only 40 nm, providing areas of potential electrical contact. The cell membranes are heavily invested with long finger-like invaginations, associated with sarcoplasmic reticular cisternae, and surface caveolae; the latter are associated with the numerous dense body membrane attachment plaques found in this muscle. The radular retractor muscle possesses a significant sarcoplasmic reticulum of peripheral cisternae and deeper vesicles associated with mitochondria. The surface caveolae may result from myofilament force exerted via attachment plaques at the cell membrane, while deeper invaginations may constitute a rudimentary transverse tubular system to relay surface depolarization to associated sarcoplasmic reticular cisternae inducing calcium release to effect excitation-contraction coupling. The radular retractor muscle possesses the usual thick paramyosin and thin actin myofilaments, the latter associated with dense bodies and attachment plaques presumably to transduce force to the cell membrane. The mitochondria are unusually large and packed into dense central clusters surrounded by large deposits of glycogen granules. The nerve endings on the radular retractor muscle fibres show four different types of transmitter vesicle, presumably related to the four kinds of agonist action in this muscle, cholinergic, serotonergic, peptidergic and purinergic. All nerve endings have mixed vesicle populations, clear evidence of co-transmission. In this muscle we see a modification of usual smooth muscle structure to effect fast sustained contractions, an ultrastructural configuration functionally designed for the muscle's central role in the feeding cycle.Abbreviations ABRM anterior byssus retractor muscle - EC coupling excitation-contraction coupling - RP radular protractor muscle - RR radular retractor muscle - SR sarcoplasmic reticulum - T-system transverse tubular system     

Feeding physiology of the carnivorous gastropod Thais clavigera (Kuster): do they eat “soup”?

The ability to feed on suspended and dissolved organic nutrients may have been retained in predatory gastropods during evolution. The carnivorous muricid neogastropod Thais clavigera feeds on prey by boring through their shells, followed by extracellular digestion and suction of the nutrient-rich fluid of the prey's body tissues. This study reports on the effect of feeding on suspended and soluble organic nutrients (SSONs) on the survival, growth, and various physiological activities including scope for growth and glycogen stores of T. clavigera. Juvenile T. clavigera of similar shell length (23.8±1.7 mm) were either starved, fed with mussel Septifer virgatus, fed with SSONs from homogenized mussel flesh (S. virgatus), or fed with both mussels and SSONs, and kept in artificial seawater (salinity: 30‰) for 50 days. Ingestion of SSONs by the animals was significant. Feeding with the “soup” (i.e., SSONs) reduced tissue wastage and improved condition index of the snails. T. clavigera fed in this manner were intermediates between the starved and the mussel-fed groups in terms of mortality, growth, food consumption, respiration, scope for growth, and glycogen content measurements. Furthermore, T. clavigera fed with both mussels and SSONs exhibited an identical energy requirement and similar values of various physiological measurements as that of those fed solely on mussel flesh. Feeding of SSONs contributed >10% of the overall energy requirement when both SSONs and mussel prey were available. The results indicate that energy from suspended and dissolved organic nutrients can contribute to the maximization of energy input in T. clavigera, which may favor better survivorship and thus lifetime fitness.