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Biomedical subjects

G Smagghe

Publications and source records attributed to G Smagghe.

At least 19 recordsLinked to original sources

Growth and mitogenic effects of arylphorin in vivo and in vitro.

In insects, developmental responses are organ- and tissue-specific. In previous studies of insect midgut cells in primary tissue cultures, growth-promoting and differentiation factors were identified from the growth media, hemolymph, and fat body. Recently, it was determined that the mitogenic effect of a Manduca sexta fat body extract on midgut stem cells of Heliothis virescens was due to the presence of monomeric alpha-arylphorin. Here we report that in primary midgut cell cultures, this same arylphorin stimulates stem cell proliferation in the lepidopterans M. sexta and Spodoptera littoralis, and in the beetle Leptinotarsa decemlineata. Studies using S. littoralis cells confirm that the mitogenic effect is due to free alpha-arylphorin subunits. In addition, feeding artificial diets containing arylphorin increased the growth rates of several insect species. When tested against continuous cell lines, including some with midgut and fat body origins, arylphorin had no effect; however, a cell line derived from Lymantria dispar fat body grew more rapidly in medium containing a chymotryptic digest of arylphorin.

Animals↗

Presence of angiotensin converting enzyme isoforms in larval lepidoptera (Spodoptera littoralis).

In this research the presence of angiotensin converting enzyme (ACE) in larvae of the lepidopteran Spodoptera littoralis was evaluated. Making use of the substrate Abz-FRK-(Dnp)P-OH and the specific inhibitor captopril at 10 microM, ACE activity was determined in a fluorescence assay for intact larvae, hemolymph, head, midgut and dorsal tissue. In dorsal tissue and hemolymph, ACE activity was highest. These data are consistent with a possible role for ACE in contractions of the dorsal vessel and metabolism of circulating peptide hormones in the hemolymph. After the presence of ACE was confirmed, a sequential procedure of anion exchange and size exclusion chromatography was applied to purify ACE from whole wandering larvae (last stage). With this procedure, three different ACE pools were collected that cleaved the fluorogenic substrate Abz-FRK-(Dnp)P-OH. Activity could be inhibited by a final concentration of 2.5 microM captopril. In addition, two out of three samples eluted at different salt concentration and thus ACE 1, 2 and 3 represent at least two different ACE isoforms. These data reveal that ACE is present in S. littoralis and that at least two out of three isolated ACE forms are truly isoforms.

Angiotensin-Converting Enzyme Inhibitors↗

ACE inhibitor captopril reduces ecdysteroids and oviposition in moths.

By using the selective ACE inhibitor captopril, we studied the effect of the angiotensin converting enzyme (ACE) on larval growth, metamorphosis, and reproduction in a lepidopteran species, the cotton leafworm, Spodoptera littoralis. Captopril was detrimental to adult formation and oviposition, and in female moths it elicited decreasing ecdysteroid levels, but increasing trypsin activities. Our results suggest that captopril downregulates oviposition by two independent pathways. Apparently, oviposition is influenced by a complex interaction of ACE, trypsin activity, and ecdysteroid levels.

Age Factors↗

Comparative toxicity of three ecdysone agonist insecticides against the Mediterranean flour moth.

The Mediterranean flour moth, Ephestia Kuehniella Zeller (Lepidoptera: Pyralidae), is an important pest in stored products worldwide, and is one of the major pests in flour mills in Algeria. Because environmental consideration, alternative approaches to neurotoxic insecticides, as well as safe, effective, and sound integrated pest management strategies are developed pest control agents such as the insect growth regulator (IGRs). Among these IGRs, the bisacylhydrazine derivatives are nonsteroidal ecdysterold agonists that mimic the action of moulting hormones and induce a precocious and incomplete moult in several insect orders. In topical bioassays using the pupae of E. kuehniella, three ecdysteroid agonists: RH-5849, the first bisaclhydrazine ecdysone agonist and two analogs, RH-5992 (tebufenozide) and RH-0345 (halofenozide), were evaluated on the reproduction under laboratory conditions. In a first series of experiments, the efficacy of these compounds was tested. These compounds exhibited insecticidal activity and the duration of pupal development was reduced with a dose-response relationship. Among the three tested compounds, tebufenozide (LD50 = 0.005 microg) appeared the most potent ecdysteroid agonist against E. kuehniella (RH-5849: LD50 = 0.05 microg and RH-0345: LD50 = 5.10 microg). In a second series of experiments, the effects of the ecdysone agonists (LD50) were investigated on the reproduction. Data showed that the three compounds affected growth of ovaries as evidenced by morphometric measurements of the ovaries from newly emerged adult females. In addition, the thickness of the chorion from basal oocytes was reduced only by RH-5992 and RH-0345. However, electron microscopic observations revealed that the three compounds had no significant effect on the fine structure of chorion. Finally, measurements of ovarian ecdysteroids' production by an enzyme immunoassay showed an increase in the hormonal amounts recorded in treated series compared to control series.

Animals↗

Activity of kk-42 in combinated treatment with RH-0345 or 20-hydroxyecdysone on morphometric measurements and free ecdysteroid in eggs of mealworms.

RH-0345 (halofenozide), a bisacylhydrazine derivative, is a nonsteroidal ecdysteroid agonist that mimics the action of the moulting hormones, while KK-42, an imidazole compound, is a potent inhibitor of ecdysteroid biosynthesis. Recent experiments with Tenebrio molitor suggested that the reduction of ecdysteroid titer, leading to a reduction of reproductive capacity, is due to a direct and rapid action of KK-42 on ecdysterold biosynthesis. Moreover, RH-0345 could partly restore the effects on reproductive events induced by KK-42. On the other hand, RH-0345 was found to affect growth and development of ovaries in a manner similar to that 20-hydroxyecdysone (20-E) and increased the ecdysteroid production. Therefore, the present study evaluates KK-42 applied topically (10 microg/insect) in combination with RH-0345 or 20-E in order to obtain more information on the mode of action of this compound on reproduction in T. molitor. In a first series of experiments, the compounds were assayed on morphometric measurements of freshly laid eggs. Results showed that both the weight and the volume of eggs increase significantly in series treated by KK-42 followed by 20-hydroxyecdysone as compared to controls and treated series by KK-42 followed by RH-0345. Data from enzyme immunoassay measurements revealed that KK-42 applied before 20-E was found to reduce significantly the amounts of free ecdysteroids in eggs comparatively to the others series.

Animals↗

The angiotensin converting enzyme inhibitor captopril reduces oviposition and ecdysteroid levels in Lepidoptera.

The role of angiotensin converting enzyme (ACE, peptidyl dipeptidase A) in metamorphic- and reproductive-related events in the Egyptian cotton leafworm, Spodoptera littoralis (Lepidoptera, Noctuidae) was studied by using the selective ACE inhibitor captopril. Although oral administration of captopril had no effect on larval growth, topical administration to new pupae resulted in a large decrease of successful adult formation. Oviposition and overall appearance of adults emerging from treated larvae did not differ significantly from those emerging from non-treated larvae. In contrast, topical or oral administration of captopril to newly emerged adults caused a reduction in oviposition. By evaluating the effect of captopril on ecdysteroid titers and trypsin activity, we revealed an additional physiological role for ACE. Captopril exerted an inhibitory effect on ecdysteroid levels in female but not in male adults. Larvae fed a diet containing captopril exhibited increased trypsin activity. A similar captopril-induced increase in trypsin activity was observed in female adults. In male adults, however, captopril elicited reduced levels of trypsin activity. Our results suggest that captopril downregulates oviposition by two independent pathways, one through ecdysteroid biosynthesis regulation, and the other through regulation of trypsin activity. Apparently, fecundity is influenced by a complex interaction of ACE, trypsin activity, and ecdysteroid levels.

Angiotensin-Converting Enzyme Inhibitors↗

The hemolymph of caterpillars Spodoptera littoralis: physico-chemical properties and ionic composition compared to culture media.

In this paper, we determined some physico-chemical properties like osmotic pressure, pH and electrical conductivity of the hemolymph from caterpillars of Spodoptera littoralis (Lepidoptera: Noctuidae) during the last larval instar. It was of interest that we observed an increase in osmotic pressure with the increase in age in the last instar that may concur with the start of histolysis at metamorphosis. These physicochemical properties were then compared to those of Grace's and modified Grace's tissue culture medium. In addition, concentrations of the cations Na, K, Ca and Mg, and the anions Cl, NO3, PO4 and SO4 were determined in the insect hemolymph of S. littoralis. The cations K and Mg reached high values with a percent of about 52% of the total amount of cations. The concentration of sodium was low. The total sum of the anions consisted about 56 meq/1, and this allows to neutralise about 45 % of the total cations.

Animals↗

Reproductive effects in German cockroaches by ecdysteroid agonist RH-0345, juvenile hormone analogue methoprene and carbamate benfuracarb.

Blatta germanica is the more prevalent cockroach species in Algeria. In the present study, we tested the effect on reproduction in B. germanica of two insect growth regulators, RH-0345, a benzoylhydrazine analogue that mimics the action of 20-hydroxyecdysone, and methoprene, one of the most commercially important juvenile hormone analogues, and a novel carbamate insecticide, benfuracarb. The compounds were applied topically (10 and 20 microg/insect for RH-0345, and 1 and 10 microg/insect for methoprene) or orally administrated (at 2% for benfuracarb) on newly emerged females and evaluated on reproductive events during the adult life (2, 4 and 6 days). Treatment with RH-0345 and benfuracarb reduced significantly the number of oocytes, the size and the volume of the basal oocyte during the experimental period. Methoprene distorted the ovarian development since it caused a significant reduction in the number of oocytes at 2, 4 and 6 days for the two tested doses, and an increase in oocyte size at 2, 4 and 6 days with 1 microg and a decrease with 10 microg. In a second series of experiments, the effects of these compounds were assayed on the ovarian proteins. Data from biochemical analysis revealed that RH-0345 and benfuracarb reduced the ovarian amounts of proteins, while treatment with methoprene increased it during the sexual maturation.

Administration, Topical↗

In vitro effects of RH-0345 and KK-42 on ecdysteroid levels and cuticle synthesis by pupal integument of mealworms.

In this study, the effects of two insect growth regulators (KK-42 and RH-0345), applied either alone or in combination, were evaluated on the ecdysteroid production in vitro. Integument explants from the abdominal sternites of newly ecdysed pupae of mealworms, Tenebrio molitor, were cultured and the amounts of ecdysteroids, released into the culture medium, were determined by an enzyme immunoassay (EIA) after various intervals of incubation. In combined treatments, explants were cultured for the first two days in a medium added with the first compound and then transferred in new medium containing the second compound. EIA measurements showed that RH-0345, either alone or followed by KK-42, resulted in higher amounts of ecdysteroids as compared to controls. In contrast, KK-42 alone caused a significant reduction. But, when KK-42 was followed by RH-0345, the ecdysteroid amounts were equal to controls. In another series of experiments, the effects on cuticle secretion were examined. Only in the case that explants were treated with RH-0345, either alone or followed by KK-42, new cuticle synthesis was observed.

Analysis of Variance↗

Topical toxicity of two acetonic fractions of Trichilia havanensis Jacq. and four insecticides to larvae and adults of Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae).

The toxicity of botanical origin compounds such as two acetonic fractions of the seed kernels of the Meliacea Trichilia havanensis Jacq with insecticide properties (azadirone (F12) and the mixture F18 [1,7+3,7-di-O-acethylhavanensin (4:1)], three insecticides commercially available (imidacloprid, natural pyrethrins+PBO, triflumuron) and phloxine B, were tested in the laboratory. Topical bioassays using third instar and newly emerged adults of the lacewing Chrysoperla carnea (Stephens) at the maximum field recommended rate in Spain for commercials and at 1,000 ppm of active ingredient for T. havanensis acetone fractions and phloxine-B, were carried out. Imidacloprid and triflumuron were very toxic to third instar larvae inhibiting adult emergence, being the rest of insecticides harmless. Fecundity and fertility were not affected by the non-toxic compounds. Concerning adults, only imidacloprid and natural pyrethrins killed them significantly 24 hours after treatment. Phloxine B, triflumuron and T. havanensis fractions were harmless and did not cause any effect on fecundity and fertility with the exception of triflumuron, which reduced considerably the egg hatch. It can be concluded that T. havanensis acetonic fractions and phloxine B were non-toxic to larvae and adults of C. carnea when treated topically, whereas triflumuron, natural pyrethrins and imidacloprid affected one or more of the evaluated parameters under our conditions.

Acetone↗

Comparative effects of some ecdysteroid agonists in mealworms: ecdysteroid amounts and protein analysis in developing ovaries under in vivo conditions.

Some ecdysteroid agonists (RH-0345, RH-5849, RH-5992) were tested topically (10 microg/female) on newly emerged adult females of mealworms under in vivo conditions. In a first series of experiments the compounds were assayed on ovarian ecdysteroid production measured at day 2 and 4 following treatment, corresponding to the beginning and the end of vitellogenesis. All tested compounds resulted in a significant increase in ovarian ecdysteroid amounts. RH-5992 and RH-0345 were more active than RH-5849. In a second series of experiments, the activity of the compounds was investigated on ovarian proteins. The ecdysteroid agonists caused a significant reduction of protein amounts in the ovaries, and electrophoretic separation of ovary extracts from treated series showed that one or two bands were missing. These observations may an interference with the vitellogenesis process.

Animals↗

Action of brassinosteroids in the cotton leafworm Spodoptera littoralis.

The two native plant hormones 24-epibrassinolide and 24-epicastasterone showed 50% competition for binding at IC(50) of 1-3.6 microM with [(3)H]ponasterone A using cultured imaginal wing discs from last-instar larvae of the cotton leafworm, Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae). However, culture of imaginal wing discs in different concentrations of brassinosteroids, even up to 100 microM, demonstrated no induction of evagination. In contrast, 20E and the non-steroidal agonist RH-5992 competed respectively about 23- and 42-fold more effectively with labeled ponasterone A, and their ability (EC(50)) to induce disc evagination in vitro was 158 and 87 nM, respectively. Injection of 10 microg of brassinosteroids in newly-moulted last-instar larvae did not cause mortality above controls; higher mortalities were scored when brassinosteroids were injected late in the last instar.

Animals↗