Phytoecdisteroids from Serratula coronata when growing ducklings.

This article presents the results of comprehensive studies to analyze the effect of a mixture of phytoecdysteroids extracted from the juice of Serratula coronata L. on the productivity and vitality of ducklings when grown for meat, and the optimal doses of its inclusion in the diet of the bird are revealed. The methodological basis of this study was the earlier works of domestic and foreign scientists on the topic under study. In the studies, a mixture of ecdysteroids extracted from the juice of the Serratula coronata L. was used according to the method developed by a team of scientists of the Ufa Federal Research Center of the Russian Academy of Sciences (Patent RU 2151598). The object of the study was the young ducks of the cross breed "Agidel 34" of the Beijing breed. It was established that the use of phytoecdysteroids in the diets of ducklings at a dose of 1.0 mg/l of drinking water allowed to increase the safety of the livestock by 4.0%, live weight by 4.5% (p <  0.01), average daily live weight gain by 3.0-3.5%, gutted carcass weight - 7.1%. At the same time, feed costs per unit of production decreased by 2.0%, and the profitability of duck meat production increased by 5.2%.

Characterization and expression patterns of key ecdysteroid biosynthesis and signaling genes in a spider mite (Panonychus citri).

Ecdysteroids play a crucial role in regulating molting in the phylum of Arthropoda and much is known with members of the subphylum of Hexapoda including the Insecta. However, this is still unclear in key pests as spider mites belonging to the subphylum of Chelicerata that originated earlier in the Cambrian period. In this study, we investigated 14 key genes of ecdysteroid biosynthesis and signaling and their expression over the different developmental stages in the citrus red mite, Panonychus citri (Acari: Stigmaeidae). P. citri is an economically important and widespread pest of citrus crops and it has five developmental stages of egg, larva, protonymph, deutonymph and adult. Typically, the expression of the ecdysteroid-synthesizing Halloween gene Spook (PcSpo) followed a positive zigzag-like pattern with a peak in the first half of each developmental stage and a drop in the second half prior to the molting to the next stage. Similar to PcSpo, PcDib, PcSad, PcRXR2, PcE75 and PcHR38 showed a positive zigzag-like expression pattern, while that of PcE78, PcHR3 and PcFTZ-F1 was opposite that we called a negative zigzag-like pattern. Silencing of the PcSpo gene by RNAi showed that molting was inhibited. Interestingly, we could rescue these RNAi effects by supplementing ponasterone A (PonA) and not by 20E, which is indicative that mites use PonA rather than 20E as ecdysteroid hormone. Modeling of the ecdysteroid receptor (PcEcR) hormone binding cavity also predicted binding of PonA, but showed a steric hindrance for 20E. We believe our data provide insight into the evolution and expression patterns of key ecdysteroid biosynthesis and signaling genes in a distant, non-insect species, and can become a foundation to develop new targets for controlling important agricultural pests such as spider mites.

BBB penetration-targeting physicochemical lead selection: Ecdysteroids as chemo-sensitizers against CNS tumors.

The anticancer potential of ecdysteroids, especially their chemo-sensitizing activity has recently gained a substantial scientific interest. A comprehensive physicochemical profiling was performed for a set of natural or semi-synthetic ecdysteroids (N=37) to identify a lead compound against central nervous system (CNS) tumors. Calculated properties, such as lipophilicity (clogP), topological polar surface area (TPSA), brain-to-plasma ratio (clogBB) along with the measured blood-brain barrier specific in vitro permeability (logPe) were evaluated in parallel. Compounds with the highest CNS-availability predicted (clogBB>0.0 and logPe>-6.0) showed moderate to high lipophilicity (clogP=3.89-5.25), relatively low TPSA (94.45Å2), and shared a common apolar 2,3- and 20,22-diacetonide motif (25, 30-33). These ecdysteroids were selected for testing their capacity to sensitize SH-SY5Y neuroblastoma cells to vincristine. All of the five tested compounds exerted a remarkably strong, dose dependent chemo-sensitizing activity: at 2.5-10.0µM ecdysteroids increased the cytotoxic activity of vincristine one to three orders of magnitude in (e.g., from IC50=39.5±2.9nM to as low as 0.056±0.03nM). Moreover, analysis of the combination index (CI) revealed outstanding synergism between ecdysteroids and vincristine (CI50=0.072-0.444). Thus, based on drug-likeness, physchem character and in vitro CNS activity, compound 25 was proposed as a lead for further in vivo studies.

[Condition of protective intestinal microbiota populations under stress exposure in rats received different diets with bioactive food components].

The evaluation of the levels of major colon microbiota populations (lactobacilli, bifidobacteria, enterobacteria) was carried out in two 15-days experiments on Wistar rats, exposed to stress factor (electric shock) and fed with different diets with the addition of biologic active micronutrients [extract from the leaves of Serratula coronata L. and Enzymatic hydrolyzate of the mussels meat (EHMM)]. In the first experiment animals were fed with a common vivarium diet. In the experimental group the water extract from leaves of Serratula coronata L. as a phytoecdysteroid source (5 mg per 1 kg body weight) was added to water. In the second experiment rats received balanced semisynthetic diet. In the diet of the experimental group the part of the protein (casein) was replaced by the peptides from EHMM. During the experiment the animal body weight was measured. On the 14th day of the experiment the animals were subjected to stress stimulation [electrodermal stimulation on paws (electric current 0.4 mA for 8 seconds)]. On the last day of the experiment the animals were euthanized by decapitation and micro-ecological research of protective microbiota populations in the cecal contents was carried out. The relative body weight increase was recorded in both experiments. In the second experiment in animals receiving EHMM this index (68.2 ± 3.0%) was considerably higher than in the control group and in the experimental group receiving no EHMM (57.2 ± 4.0 and 59.7 ± 2.8% respectively). The results of the microecological study showed different effect of diets with biologically active micronutrients on the population levels of lactobacilli. In the experiment with common vivarium diet no significant changes of the levels of the studied colon microbiota populations had been recorded in the rats of control group compared with rats of experimental group, exposed to stress factor but received no extract from Serratula coronata L. The decrease of the levels of lactobacilli by the end of the experiment was observed in the experimental group of rats received water extract from the leaves of Serratula coronata L (content of lactobacilli 7.76 ± 0.17 lg CFU/g) compared to those in control group and experimental group of rats received no extract (8.4 ± 0.09 and 8.69 ± 0.07 lg CFU/g respectively). Feeding with the balanced semisynthetic diet with the addition of EHMM or without it had a positive effect on the levels of lactobacilli and their balance with the aerobic component of the Enterobacteriaceae. There was a trend toward increased levels of lactic acid bacteria in the experimental group received EHMM (9.16 ± 0.12 lg CFU/g) compared with the contents in the control group and in the experimental group exposed to stress factor without adding EHMM in the diet (8.74 ± 0.34 and 8.79 ± 0.23 lg CFU/g, respectively). The conclusion about the positive (protective) effect of a semisynthetic diet enriched with peptides from EHMM was made based on the comparison of indicators that reflect the status of non-specific resistance of the organism: the integral criterion of weight gain and the levels of major colon microbiota populations of laboratory animals.

[Biochemical parallels of cellular adaptive reactions at chronic low-intensity irradiation and action of phitoecdysteroid preparation serpisten].

Comparison of action of chronic γ-irradiation at a dose of 22.6 cGy and the serpisten substance containing phitoecdysteroids at small doses of 5 and 50 mg/kg on biochemical indicators in erythrocytes and tissues of white not purebred mice is given. It is established that in both cases there is an increase of minor fractions of cardiolipin and phosphatidic acid, lysophosphatidilcholin and a share of phospholipids as part of common lipids. Course administration of serpisten to rats at the total doses of 12 and 30 mg/kg leads to an increase in tissues of thermal shock proteins of family 70 (Hsp70 and Hsc70). Similarity of action of ecdysteroid preparations and the influence of stress factors of physical nature of low intensity (gamma radiation at a small dose) have been detected in mice, which manifest themselves in some chain links of lipid peroxidation processes as well as an increase in biosynthesis of thermal shock proteins of family 70 (Hsp70 and Hsc70) in rats at administration of serpisten.

Study of behavioral parameters during learning of rats in an operant feeding task and evaluation of biochemical indexes after dietary consumption of the phytoecdysteroid extract.

Experiments on rats were performed to study the process of operant feeding learning, locomotor activity, oxygen consumption, and concentrations of corticosterone, ß-endorphin, and prostaglandin E in blood serum after dietary treatment with the phytoecdysteroid extract. Administration of phytoecdysteroids was followed by the improvement of learning and increase in oxygen consumption and locomotor activity. Locomotor activity and oxygen consumption in the majority of control rats and phytoecdysteroid-treated animals were shown to be interrelated with the total locomotor activity and goal-directed operant behavior, respectively. No significant differences were found in the concentration of hormones in blood serum from animals of the control and treatment groups.

The metabolism of 20-hydroxyecdysone in mice: relevance to pharmacological effects and gene switch applications of ecdysteroids.

Ecdysteroids exert many pharmacological effects in mammals (including humans), most of which appear beneficial, but their mechanism of action is far from understood. Whether they act directly and/or after the formation of metabolites is still an open question. The need to investigate this question has gained extra impetus because of the recent development of ecdysteroid-based gene-therapy systems for mammals. In order to investigate the metabolic fate of ecdysteroids in mice, [1α,2α-(3)H]20-hydroxyecdysone was prepared and injected intraperitoneally to mice. Their excretory products (urine+faeces) were collected and the different tritiated metabolites were isolated and identified. The pattern of ecdysteroid metabolites is very complex, but no conjugates were found, in contrast to the classical fate of the (less polar) endogenous vertebrate steroid hormones. Primary reactions involve dehydroxylation at C-14 and side-chain cleavage between C-20 and C-22, thereby yielding 14-deoxy-20-hydroxyecdysone, poststerone and 14-deoxypoststerone. These metabolites then undergo several reactions of reduction involving, in particular, the 6-keto-group. A novel major metabolite has been identified as 2ß,3ß,6α,22R,25-pentahydroxy-5ß-cholest-8(14)-ene. The formation of this and the other major metabolites is discussed in relation to the various effects of ecdysteroids already demonstrated on vertebrates.

[Biological effects of phytoecdysteroids and chronic low-dose irradiation].

The influence of serpistene in dose of 5 and 50 mg/kg on chronic low-dose gamma-irradiation (22.6 cGy) effects on cytogenetic (abnormal sperm cell, marrow bone micronucleus) and function and morphology (thyroid and adrenal glands) parameters of mice was estimated. The serpistene modifies effects of gamma-irradiation depends on the administration regime and a dose of the substance. The most expressive radioprotective effect on endocrine organs after serpistene prophylactic administration was found. The prophylactic dose was 5 mg/kg for adrenal gland and both doses--for thyroid gland. The most expressive radioprotective effect on marrow bone cells after serpistene therapeutic administration in a dose of 5 mg/kg was found. The most expressive antimutagenic effect on somatic and germinal cells of prophylactic and therapeutic administration in a dose of 50 mg/kg was found.