Hemolymph, removal

Small and Thacker 1994). So, the ability to detect chemicals released by dead conspecifics and to associate them with the potential for a new shell has been conserved in the transition from sea to land the attractant is contained in the crab hemolymph, its power being displayed immediately upon the hemolymph removal without the need of any proteases, as required with snail odors. A recent gas chromatography-mass spectrometry study (Schmidt et al. 2009) showed that it is the volatile alcohol 3-decanol found in the hemolymph of Clibanarius vittatus that stimulates shell investigation behavior in conspecifics. 

Attractive Compounds. The female-produced sex pheromone of the yellow mealworm beetle, Tenebrio molitor, is (R)-4-methyl- 1-nonanol [316] 163 (Scheme 18). Careful investigations on the biosynthesis of this compound [317] revealed that it is produced through a modification of normal fatty acid biosynthesis (Fig. 1, Fig. 2) propanoate serves as the starter, while formal chain elongation with acetate, propanoate, and acetate (accompanied by removal of the oxygens) produces 4-methylnonanoate which yields the pheromone alcohol after reduction. The structures and role of proteins that are present in the hemolymph or secreted by the tubular accessory glands of T. molitor, and that may carry lipophilic chemical messengers (like pheromones) are under investigation [318,319]. 

Dry the target under gentle vacuum and wash (also used for desalting if cmde hemolymph or digestion mixture) the sample preparation twice with 1 piL of 1 % TFA. Remove the washing solution after a few seconds using forced air, and dry the sample under vacuum. 

To verify in vitro results, we have also developed an in vivo assay for L. dispar PTTH (52, Thyagaraja et al, in preparation). This assay is comparable to larval assays develop for Af. sexta (56 and B. mori (58) and uses last instar female larvae. Neck ligation prior to day 7, the day when a small pre-peak in hemolymph ecdysteroid titer appears, blocks further development. Injection of PTTH-containing extracts posterior to the ligation, including post-embryonic egg extract, reinitiates development and subsequent pupation. Attempts to develop an in vivo assay comparable to the pupal-adult B. mori assay (5B were unsuccessful, since brain removal as close as 15 min post-pupal ecdysis failed to block L. dispar female adult development (Thyagaraja et a/., unpublished results). 

In diapausing females of the Savio strain of L. migratoria. the basal activity of the CA is totally inhibited in vivo, as established by the arrest of oocyte development and absence of JH in the hemolymph (27). However, these same glands are immediately activated when they are incubated in vitro (i. e., denervated and removed from their natural milieu), indicating that their maturation has not been retarded. In vivo denervation of the glands (NCA-I transection) in diapausing females elicits elevated JH titer and induces oocyte development, indicating that this short-term inhibition is axonally imposed. 

The function of apoLp-IIl is to facilitate transport of lipid from sites of storage in the fat body to sites of utilization in certain metabolic situations, e.g., flight. The triacylglycerol stores of the fat body are converted to DG, which leaves the fat body and becomes associated with preexisting HDLp in the hemolymph. In the process, HDLp is converted to LDLp and several molecules of apoLp-III become associated with LDLp. LDLp moves to the flight muscle, where the DG is hydrolyzed by a lipoprotein lipase. As the DG is removed, LDLp is converted back to HDLp and apoLp-III dissociates. The HDLp and apoLp-III then cycle back to the fat body to carry more DG (see Section V for details). 

Eggs from Hyalophora cecropia (Telfer, 1960), Samina cynthia (Chino et al., 1977), M. sexta (Kawooya et al., 1988), and R. prolixus (Gondim et al., 1989b) have been shown to contain a very high-density lipophorin, VHDLp-E, which is derived from HDLp-A in the hemolymph by a receptor-mediated process (Kawooya et al., 1988 Telfer and Pan, 1988 Kulakosky and Telfer, 1990) this represents the only known exception to the generalization that lipophorin delivers its lipids to tissues without internalization. The conversion of HDLp-A to VHDLp-E involves removal of DG, which is catalyzed by a lipoprotein lipase found in the yolk body of the egg (Van Antwerpen and Law, 1992). However, in spite of the presence of lipophorin in the egg, 90% of the lipid in the egg is... 

Gabacullne was injected at 1000 yg/g (=35 mM hemolymph concentration) ethyl 4-aminooxybutyrate at 12.5 yg/g, half the LD50 (=0.4 mM hemolymph concentration). Cockroaches were then injected with [14c(U)]GABA (79,600 dpm, abdominal route) and placed in closed flasks containing 0.2 ml N KOH as a CO2 trap. At 2, 5, and 20.5 h the traps were removed for measurement of 1 C02. 

The transferrins are a class of iron-binding glycoproteins which have been found in the blood serum of a variety of vertebrates and are presumably also present in moth hemolymph [see (74) and references therein], These proteins would appear to mediate the absorption and distribution of iron (75), and could play some role in the movement of carbon dioxide in the body (76). Proteins of similar characteristics, lactoferrin and conalbumin, have been isolated from milk and avian egg white respectively. It has been proposed (77) that conalbumin could prevent bacterial contamination of the egg yolk by removing free iron. A similar bacteriostatic action could be performed by lactoferrin in milk, and it is possible that this protein is involved in controlling the intestinal flora in infants (78). 

Fig. 21.6 Effects of emersion on hemolymph CHH (circles, solid line). Lobsters were removed from the water and placed in an incubator at ambient temperature (13.0°C). Hemolymph was sampled from these animals at various time points. Controls were matched siblings that were left immersed at 13°C and sampled at the same time points. Lobsters ranged in wet weight from 100 to 145 g. Means standard deviations are shown. Control data are represented by the triangles and dashed line. Asterisks indicate significant differences from immersed controls at P < 0.01 (double asterisk) and atP < 0.001 (triple asterisk). Modified from Chang, Keller, and Chang (1998) with permission from Elsevier...

For detection and for removal of linamarin from foods (Phaseolus, Cassava) and animal fodder (flaxseed) by enzymatic cleavage and extraction processes, see Lit.. Linamarin and lotaustralin occur in the hemolymph of caterpillars of the butterfly Zygaena trifolii. ... 

Pharmacokinetics of JHE. When high titers of purified recombinant JHE from K virescens were injected into M, sexta, a rapid decline of this JHE in the hemolymph was noted (61), JHE was removed by a saturable process, possibly by receptor mediated endocytosis, and there was no difference in clearance rate between different larval stadia of M, sexta and H. virescens. As noted above, it is possible that the recombinant virus-expressed JHE may be removed before having a significant biological effect. However, baculovirus expressed JHE reaches very high activity levels in vivOy up to 20 or 40 times the maximum titers seen during the course of normal larval development in some cases (Table II 51). Despite this fact, precocious development of larvae infected with the various recombinant viruses is not seen. 

Young adult flies are decapitated, the wings removed, and the body mounted on a pin through the thorax (Kernan et al. 1994). One electrode is placed over the cut end of a thoracic bristle (humeral or anterior notopleural macrochaete bristle). A second reference electrode is inserted into the thorax so that it contacts the hemolymph. 

---