Pharmacophore based approach to design inhibitors in Crustaceans: An insight into the molt inhibition response to the receptor guanylyl cyclase.

The first set of competitive inhibitors of molt inhibiting hormone (MIH) has been developed using the effective approaches such as Hip-Hop, virtual screening and manual alterations. Moreover, the conserved residues at 71 and 72 positions in the molt inhibiting hormone is known to be significant for selective inhibition of ecdysteroidogenesis; thus, the information from mutation and solution structure were used to generate common pharmacophore features. The geometry of the final six-feature pharmacophore was also found to be consistent with the homology-modeled MIH structures from various other decapod crustaceans. The Hypo-1, comprising six features hypothesis was carefully selected as a best pharmacophore model for virtual screening created on the basis of rank score and cluster processes. The hypothesis was validated and the database was virtually screened using this 3D query and the compounds were then manually altered to enhance the fit value. The hits obtained were further filtered for drug-likeness, which is expressed as physicochemical properties that contribute to favorable ADME/Tox profiles to eliminate the molecules exhibit toxicity and poor pharmacokinetics. In conclusion, the higher fit values of CI-1 (4.6), CI-4 (4.9) and CI-7 (4.2) in conjunction with better pharmacokinetic profile made these molecules practically helpful tool to increase production by accelerating molt in crustaceans. The use of feeding sub-therapeutic dosages of these growth enhancers can be very effectively implemented and certainly turn out to be a vital part of emerging nutritional strategies for economically important crustacean livestock.

Study of in vitro endotoxin neutralization effect and antibacterial activity of limulus anti-lipopolysaccharide factor simulated peptide REMP2 / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the role of REMP2 derived from limulus anti-lipopolysaccharide factor in neutralizing endotoxin in vitro and its antibacterial activity.</p><p><b>METHODS</b>(1) REMP2 and PMB in the concentrations of 100.00, 10.00, 1.00, 0.10, 0.01 micromol/L were respectively mixed with LPS (lEU/mL), with PMB as positive control. The LPS concentrations in different specimens were determined by routine method, and the neutralizing percentage was respectively calculated. (2) After adding isotonic saline (NS), the final concentrations of REMP2 and PMB were 10, 20, 40, 80 micromol/L, and the concentration of LPS was 100 microg/L. The murine monocytic macrophages were stimulated with LPS, then cultured with REMP2 and PMB, with NS in culture as negative control. The content of tumor necrosis factor (TNF)-alpha was determined by ELISA kit. (3) The morphologic changes of Escherichia coli. was observed under electron microscope at 10, 20 and 40 minutes after addition of REMP2 to Escherichia coli suspension (with terminal concentration of REMP2 at 40 micromol/L).</p><p><b>RESULTS</b>There were no significant difference in endotoxin-neutralizing percentages between PMB and REMP2 in concentrations of 0.10, 10.00, 100.00 micromol/L (P > 0.05). The contents of TNF-alpha were 1175 +/- 162, 859 +/- 122, 645 +/- 142, 489 +/- 102 ng/L, respectively,after treatment of 10, 20, 40, 80 micromol/L REMP2, which were obviously lower than that of NS (3463 +/- 218 ng/L, P < 0.01). Under transmission electron microscope, the outer and interior membranes of Escherichia coli were obscure and rough, bacterial bodies were swollen with vacuoles in cytoplasm after treatment with REMP2.</p><p><b>CONCLUSION</b>REMP2 has ability of neutralizing endotoxin and also antibacterial activity.</p>

Effect of Varroa destructor on different haemocyte count, total haemolymph protein on larvae, pupae and adults of Apis mellifera drones

The effect of infestation with Varroa destructor Oud. [Acarina: Varroidae] on the heamolymph of the different stages in honey bee drons were studied. It decreased the total haemocyte count [THC] by 47.4%, 13.04% and 65% for infested larvae, pupae and adults, respectively. Differential haemocyte count [DHC], haemocyte surface area, corpora allata surface area [CA] and total haemolymph protein [THP] were decreased also due to infestation with varroa mite

Current progress in shrimp endocrinology-a review.

One problem in aquaculture is obtaining brood because many commercially important species are incapable of spontaneous maturation under artificial conditions. Commercial shrimp hatcheries commonly use eyestalk ablation to stimulate gonadal maturation in shrimps. Research has been conducted on the inhibition of reproductive maturation by hormones originating in the eyestalk glands and on other endocrine sources (e.g.,brain, thoracic ganglion, ovary, mandibular organ, androgenic gland and Y-organs) to determine their roles. Alternate techniques for acceleration of gonad maturation through the use of synthetic hormones or neurotransmitters may benefit aquaculture. Neurohormones and neuroregulators have been shown to accelerate gonadal maturation but an effective delivery technique must be developed for use in a large-scale aquaculture operation.

Paradoxical effects of elastase inhibitor guamerin on the tissue repair of two different wound models: sealed cut and exposed tongue wounds

Innate elastase inhibitors are known to be putatively involved in the regulation of tissue inflammation by inhibiting polymorphonuclear leukocyte (PMN) derived proteinases. The aim of this study was to evaluate affects of leukocyte elastase suppression and PMN infiltration on wound healing in mouse by administering the recombinant elastase inhibitor guamerin (rEIG) in two different wound models; 1) impaired pin-punctured dorsal mucosa of anterior tongue wound, 60 mice, treated with saline containing rEIG that were fed ad libitum and 2) stable linear excisional cutaneous wound, 40 mice, covered with fibrin sealant containing rEIG. The progress of healing was analyzed by histological methods. The tongue wounds treated with rEIG became edematous around the pin-punctured tongue wound, and influx of inflammatory cells and PMN into the underlying stromal tissue were seen rapidly after wounding and peaked between 2-4 days. Whereas the control mice showed almost no wheal formation in the pin-punctured wound, a far lesser levels of PMN infiltration, and almost complete wound closure in 4 days. In the other model, the liner excisional cutaneous wound treated with fibrin sealant containing rEIG showed early wound constriction, lesser degree of inflammatory cells influx, and complete reepithelialization in 4-5 days, whereas the wound of control mice with the fibrin sealant alone showed contrary delayed reepithelialization, greater degree of inflammatory cell infiltration, and consequencial formation of greater granulation tissue at wound site. Taken together, these data suggest paradoxical effects of rEIG on the wound healing where in the wound exposed to infiltrating milieu of microorganisms in the oral cavity, the rEIG aggravates the wound healing by interfering with other innate defensive factors and extended greater flux of PMNs to inflamed wound site, while in the wound enclosed by fibrin, the rEIG accelerated wound healing by inhibiting the inflammation-generated proteases and the acute inflammatory reaction.

Research on TALF expression in Escherichia coli / 生物工程学报

The expression of cDNA encoding Tachyleus auti-lipoposaccharide (LPS) factor, which is of interest for use as a potential inhibitor of the common core subunit of Gram-negative bacterial endotoxin. First, the TALF gene was inserted into expression vectors pGEX-4T-2, pET22b and pET28a to construct recombinant expression plasmids. The recombinant plasmids were transformed to E. coli BL21 (DE3) and the expression of TALF was examined. Results show that TALF in pET22b and pET28a vectors can't be expressed. Only the fusion protein GST-TALF was expressed in E. coli BL21 existing as inclusion bodies. From 1 liter of culture, about 4mg of fusion protein GST-TALF with 91% purity was finally obtained. No apparent bactericidal activity and LPS neutralizing activity of the fusion protein GST-TALF were found. After digested with thrombin, the fusion protein GST-TALF exhibited strong bactericidal activity and LPS neutralizing activity.

Effects of crustacean hyperglycemic hormones from Carcinus maenas and Orconectes limosus on blood and muscle glucose and glycogen concentration of Chasmagnathus granulata

The effects of purified crustacean hyperglycemic hormones (CHH) from Carcinus maenas or Orconectes limosus, and of eyestalk extract of Chasmagnathus granulata on the blood and muscle glucose and glycogen concentration of Chasmagnathus granulata were investigated. Different groups of animals (at least 7 animals per group) were injected with CHH from either C. maenas or O. limosus CHH dissolved in saline (16 pmol/animal) or crude eyestalk extract of C. granulata (1 eyestalk equivalent/animal). All injections had a volume of 10 microliters. Blood and muscle glucose and glycogen concentrations were determined immediately before the injections and after 30, 60 and 120 min. CHH administration from both species, as well as eyestalk extract, resulted in marked hyperglycemia. However, their effects were different. CHH from C. maenas also caused a decrease in the glycogen concentration of blood (from 89.8 +/- 4.3 to 76.6 +/- 3.1 mg/100 ml) and muscle (from 7.9 +/- 0.8 to 4.0 +/- 0.7 mg/g) and glucose concentration of muscle (from 2.4 +/- 0.3 to 1.2 +/- 0.2 mg/g). CHH from O. limosus caused an increase of glycogen concentration of muscle (from 4.9 +/- 1.1 to 9.0 +/- 1.1 mg/g). The injection of eyestalk extract resulted also in a decrease of hemolymph glycogen (from 157.7 +/- 20.6 to 30.2 +/- 7.7 mg/100 ml). Therefore, C. granulata may have different receptors for CHH in its different tissues, and/or in the same tissue, which act through different metabolic pathways to achieve the same final result, i.e., hyperglycemia

The hemolymph of chasmagnathus granulata dana,1851 (Decapoda - grapsidae) as a target tissue ofthe crustacean hyperglycemic hormone

The effect of crustacean hyperglycemic hormone (CHH) was investigated on the hemolymph of Chasmagnathus granulata, a meso-supralitoral crab from southern Brazil. Serum glucose increased significantly (P®0.05) after incubation of total hemolymph in the presence of the eyestalk extract of a member of the same species. Also glucose uptake from blood serum, not affected by eyestalk extract (P¼0.05) was observed after incubation of total hemolymph in the presence of glucose.The results that the hemolymph may be a target tissue of CHH and that this hormone may act by mobilizing carbohydrate reserves possibly from hematocytes.

Effects of azadirachtin in Rhodnius prolixus: data and hypotheses

The effects of azadirachtin A, a tetranortriterpenoid from the neem tree Azadirachta indica J., on both development and interaction between Trypanosoma cruzi, the causative agent of Chagas' disease, and its vector Rhodnius prolixus were studied. Given through a blood meal, a dose-rsponse relationship of azadirachtin was established using antifeedant effect and ecdysis inhibition as effective parameters. A singlo dose of azadirachtin A was able to block the onset of mitosis in the epidermis and ecdysteroid titers in the hemnolymph, determined by radioimmuneassay, were too low for an induction of ecadysis. The survival of T. cruzi was also studied in R. prolixus treated with the drug. If the trypomastigotes were fed in presence of azadirachtin A the number of parasites drastically decreased. If the drug was applied after infection of the bug with T. cruzi, the parasite was still abolished from the gut. If the insect was pretreated with azadirachtin A before infection the same observation was obtained. A single dose of azadirachtin A was enough for a permanent resistance of the insect host against its reinfection with T. cruzi and for blocking the ecdysis for a long time. The effects of azadirachtin A on the hormonal balance of the host and growth inhibition of the parasite will be discussed on the basis of the present results.

Effects of proallatotoxins (precocenes) on the development and reproduction of Rhodnius prolixus: some data

Proallatotoxins, and particularly preconcenes, are exceptionally promising models for studying Rhodnius prolixus physiology and for comparison with other natural compounds with anti-hormonal activities. Effects of preconcenes on feeding, development and reproduction of R. prolixus are being detailed. The precocenes reveal significant effects on feeding, moulting cycle (inducing precocious metamorphosis and ecdysial stasis), and reproduction of these insect. The mechanism of action of proallatotoxins was discussed based on the corpus allatum cytotoxic effect and on the ecdysteroid biosynthesis in prothoracic glands and ovaries. Further studies of these compounds on R. prolixus are need and will hopefully reveal other unesplored points regarding the action of the proallatotoxins on insects.