Efficacy of dispersed fibrous carbon sorbent in treatment of malignant fungating wounds.

AIM: To evaluate the influence of dispersed fibrous carbon sorbent (DFCS) on malignant fungating wounds (MFWs) in Guerin's carcinoma-bearing rats with an assessment of wound bacterial microflora. MATERIALS AND METHODS: The study was performed on female Wistar rats inoculated subcutaneously with Guerin T8 carcinoma into the upper interscapular region. On day 20, the tumors were infected by injecting the suspension of S. aureus 8325-4. After 24 h, gauze dressings were applied daily to the wounds in the control group and DFCS in the experimental one. The state of the wounds was assessed by visual control with photo monitoring, odor control on a verbal rating scale and microbiological analysis of qualitative and quantitative characteristics of wound microbiota. RESULTS: On the 3rd day after the start of dressings, the number of S. aureus of 8325-4 strain was significantly higher in the wounds of rats in control group than experimental one (5.14 ± 0.27 vs 3.43 ± 0.33 lg CFU/ml). The total number of Staphylococci in the DFCS-treated wounds did not differ statistically from the number of S. aureus 8325-4 strain whereas in control ones were higher by an order of magnitude. On the 10th day the total number of Staphylococci and S. aureus 8325-4 in particular, remained consistently high in the wounds of the control rats, while was almost five orders of magnitude lower and represented mainly by S. aureus 8325-4 in the experimental ones. The total number of aerobic and optionally anaerobic microorganisms was significantly lower as well. A greater variety of microorganisms in the gauze-covered wounds, as well as an their increased number were accompanied by enhancement of the wound odor from "noticeable" to "strong". There was an appearance of a "barely noticeable" odor in only one animal from the experimental group. CONCLUSION: The study has demonstrated the ability of DFCS to control substantially the bacterial microflora as well as malodor of MWFs in vivo. The results obtained can contribute to solving the problem of improving the quality of palliative care for patients with malignant and other chronic wounds.

Hygiene-Eliciting Brood Semiochemicals as a Tool for Assaying Honey Bee (Hymenoptera: Apidae) Colony Resistance to Varroa (Mesostigmata: Varroidae).

Despite numerous interventions, the ectoparasitic mite Varroa (Varroa destructor Anderson and Trueman [Mesostigmata: Varroidae]) and the pathogens it vectors remain a primary threat to honey bee (Apis mellifera Linnaeus [Hymenoptera: Apidae]) health. Hygienic behavior, the ability to detect, uncap, and remove unhealthy brood from the colony, has been bred for selectively for over two decades and continues to be a promising avenue for improved Varroa management. Although hygienic behavior is expressed more in Varroa-resistant colonies, hygiene does not always confer resistance to Varroa. Additionally, existing Varroa resistance selection methods trade efficacy for efficiency, because those achieving the highest levels of Varroa resistance can be time-consuming, and thus expensive and impractical for apicultural use. Here, we tested the hypothesis that hygienic response to a mixture of semiochemicals associated with Varroa-infested honey bee brood can serve as an improved tool for predicting colony-level Varroa resistance. In support of our hypothesis, we demonstrated that a mixture of the compounds (Z)-10-tritriacontene, (Z)-8-hentriacontene, (Z)-8-heptadecene, and (Z)-6-pentadecene triggers hygienic behavior in a two-hour assay, and that high-performing colonies (hygienic response to ≥60% of treated cells) have significantly lower Varroa infestations, remove significantly more introduced Varroa, and are significantly more likely to survive the winter compared to low-performing colonies (hygienic response to <60% of treated cells). We discuss the relative efficacy and efficiency of this assay for facilitating apiary management decisions and selection of Varroa-resistant honey bees, as well as the relevance of these findings to honey bee health, pollination services, and social insect communication.

Stock-specific chemical brood signals are induced by Varroa and Deformed Wing Virus, and elicit hygienic response in the honey bee.

The health of the honey bee Apis mellifera is challenged by the ectoparasitic mite Varroa destructor, and the numerous harmful pathogens it vectors. Existing pesticide-based Varroa controls are not sustainable. In contrast, one promising approach for improved honey bee health is the breeding of hygienic bees, capable of detecting and removing brood that is parasitized or diseased. In three experiments we find evidence to support the hypothesis that stock-specific chemical brood signals are induced by Varroa and Deformed Wing Virus, and elicit hygienic response in the honey bee. By collecting, analyzing, and running bioassays involving mite-infested and control brood extracts from three honey bee breeding stocks we: 1) found evidence that a transferrable chemical signal for hygienic behavior is present in Varroa-infested brood extracts, 2) identified ten stock-specific hydrocarbons as candidates of hygienic signaling, and 3) found that two of these hydrocarbons linked to Varroa and DWV were also elevated in brood targeted for hygienic behavior. These findings expand our understanding of honey bee chemical communication, and facilitate the development of improved hygienic selection tools to breed honey bees with greater resistance to Varroa and associated pathogens.

Backbone flexibility of extended metal atom chains. Ab initio molecular dynamics for Cr3(dpa)4X2 (X = NCS, CN, NO3) in gas and crystalline phases.

Recently published static DFT and CASSCF/CASPT2 calculations depicted extremely flat Potential Energy Surfaces (PESs) for the Cr-Cr flexibility of Cr3(dpa)4X2 (X = NCS-, CN-, NO3-) extended metal atom chains (EMACs) (M. Spivak, et al., Dalton Trans., 2017, 46, 6202). We herein explore the thermal and crystal packing effects on the structure of EMACs using ab initio molecular dynamics (MD). Car-Parrinello DFT-based simulations of the isolated molecules show that thermal energy favors asymmetric arrangements of the Cr3 chain due, in part, to the bending of the axial ligands (X) and the increased X-Cr distance, both of which weaken X → Cr σ-donation. This effect is even more prominent in the crystalline phase due to the interaction between the axial ligands of neighboring molecules in the unit cell. This could explain the typical discrepancies between the experimental and theoretical characterization of Cr3 EMACs observed in the literature.

A multiconfigurational approach to the electronic structure of trichromium extended metal atom chains.

Density functional theory, Complete Active Space Self-Consistent Field (CASSCF) and perturbation theory (CASPT2) methodologies have been used to explore the electronic structure of a series of trichromium Extended Metal Atom Chains (EMACS) with different capping ligands. The study is motivated by the very different structural properties of these systems observed in X-ray experiments: the CN--capped example has a symmetric Cr3 unit while for the NO3--capped analogue the same unit has two very different Cr-Cr bond lengths. Density functional theory fails to locate an unsymmetric minimum for any of the systems studied, although the surface corresponding to the asymmetric stretch is very flat. CASPT2, in contrast, does locate an unsymmetric minimum only for the NO3--capped system, although again the surface is very flat. We use the Generalized active space (GASSCF) technique and effective Hamiltonian theory to interrogate the multi-configurational wavefunctions of these systems, and show that the increase in the σ-σ* separation as the chain becomes unsymmetric plays a defining role in the stability of the ground state and its expansion in terms of configuration state functions.

A Comparison of Honey Bee-Collected Pollen From Working Agricultural Lands Using Light Microscopy and ITS Metabarcoding.

Taxonomic identification of pollen has historically been accomplished via light microscopy but requires specialized knowledge and reference collections, particularly when identification to lower taxonomic levels is necessary. Recently, next-generation sequencing technology has been used as a cost-effective alternative for identifying bee-collected pollen; however, this novel approach has not been tested on a spatially or temporally robust number of pollen samples. Here, we compare pollen identification results derived from light microscopy and DNA sequencing techniques with samples collected from honey bee colonies embedded within a gradient of intensive agricultural landscapes in the Northern Great Plains throughout the 2010-2011 growing seasons. We demonstrate that at all taxonomic levels, DNA sequencing was able to discern a greater number of taxa, and was particularly useful for the identification of infrequently detected species. Importantly, substantial phenological overlap did occur for commonly detected taxa using either technique, suggesting that DNA sequencing is an appropriate, and enhancing, substitutive technique for accurately capturing the breadth of bee-collected species of pollen present across agricultural landscapes. We also show that honey bees located in high and low intensity agricultural settings forage on dissimilar plants, though with overlap of the most abundantly collected pollen taxa. We highlight practical applications of utilizing sequencing technology, including addressing ecological issues surrounding land use, climate change, importance of taxa relative to abundance, and evaluating the impact of conservation program habitat enhancement efforts.

Probiotic strains of lactobacilli and bifidobacteria alter pro- and anti-inflammatory cytokines production in rats with monosodium glutamate-induced obesity.

The aim of this study was to investigate the effect of probiotic strains of Lactobacillus casei IMV B-7280, Bifidobacterium animalis VKL, B. animalis VKB on the pro- and anti-inflammatory cytokines production in Wistar male rats with monosodium glutamate (MSG)-induced obesity. It was established that neonatal administration of MSG to rats leads to increasing levels of the interleukin (IL)-1ß and IL-12, and to decreasing ofthe IL-4, IL-10 and tumor growthfactor (TGF)-ß levels in the bloodserum. After administration of the B. animalis VKL - B. animalis VKB - L. casei IMV B-7280 composition to obese rats the level of the IL-lP in blood serum wasn't differ from that in the obese rats, that didn't receive of the probiotic bacteria. But there was no statistically signifcant difference comparing with intact rats. The level of the IL-12B p4O in blood serum was decreased under influence of the B. animalis VKL - B. animalis VKB - L. casei IMV B-7280 composition (18.9%, p < 0.05) and B. animalis VKL (10.5%, p < 0.05) compared with obese rats, not receiving probiotic bacteria, but remained higher than in intact animals. After administration to obese rats ofthe B. animalis VKL - B. animalis VKB - L. casei IMV B-7280 composition the levels ofthe IL-4, IL- 10 and TGF-ß increased in blood serum comparing with obese rats, not receiving probiotic bacteria. The level of the IL-10 also increased under influence of the B. animalis VKB, and IL-4 - under influence of the L. casei IMVB-7280. Our results suggest that these probiotic bacteria and probiotic composition are able to down-regulation the inflammation in rats with MSG-induced obesity but the strongest anti-inflammatory effects have probiotic composition. The ability of lactobacilli and bifdobacteria to alter the pro- and anti-inflammatory cytokines production, opens perspectives to create new treatments for obesity and metabolic syndrome based on probiotics.

OBTAINING OF THE TRANSGENIC HELIANTHUS TUBEROSUS L. PLANTS, CALLUS AND "HAIRY" ROOT CULTURES ABLE TO EXPRESS THE RECOMBINANT HUMAN INTERFERON ALPHA-2b GENE.

This work is the first to our knowledge to describe the successful attempt of Agrobacterium rhizogenes-mediated transformation of topinambour in order to obtain the transgenic H. tuberosus plants, callus and "hairy" root cultures. The plasmid vectors contained the sequence of interferon gene fused with Nicotiana plumbagenifolia L. calreticulin apoplast targeting signal driven by 35S CaMV promoter or root-specific Mll promoter. Nearly 75% isolated Ri-root lines and callus cultures were proved (by PCR analysis) to contain HuINFa-2b transgene. We also managed to obtain H. tuberosus transgenic plants through somatic embryogenesis on the transgenic "hairy" root culture. The obtained transgenic H. tuberosus cultures exhibited high-level antiviral activity that ranged from 2000 to 54500 IU/g FW that makes this crop considered a promising source of recombinant interferon alpha 2b protein.

[THE INFLUENCE OF NANODISPERSE CERIUM DIOXIDE ON ONTOGENETIC CHANGES OF ANTIOXIDANT SYSTEM IN THE MUCOSA OF THE STOMACH AND COLON IN RATS].

It was established that with age the content of lipid peroxidation products increased in the mucosa of the stomach: Diene conjugates by 30%, products which react to thiobarbituric acid by 285% and Schif bases by 181%. Nanodisperse cerium dioxide (NCD) reduced the content of lipid peroxidation in the gastric mucosa in old rats: Diene conjugates by 43 %, products which react to thiobarbituric acid by 51% and Schif bases by 44% relative to the control group of rats given age. Similarly, it was established that the content of Diene conjugates increased by 40%, products which react to thiobarbituric acid by 114% and Schif bases by 132% in the mucosa of the colon of old rats. NDC significant reduced the content of products which react to thiobarbituric acid by 69% and Schyf basics by 132%. In the stomach superoxide dismutase (by 43%) and catalase activity (by 24%) decreases with age, while in the colon superoxide dismutase activity increases (by 43%). In the colon NCD significant decreased superoxide dismutase (by 34%) and catalase activity (by 21%) relative to controls. Thus, the NDC restores lipid peroxidation in the gastric mucosa and colon, in which develops oxidative stress with age.

[Effects of mono-, poly- and composite probiotics on the ulceration caused by restraint stress].

It was studied the effect of probiotic strains of Bifidobacterium animalis VKL, Bifidobacterium animalis VKB and Lactobacillus casei IMVB-7280, and their mixtures on erosive and ulcerative lesions of the gastric mucosa (GM) of rats. GM was induced by water-immersion restraint stress. It was found that investigated probiotics did not have gastroprotective properties under a single and seven-day prophylactic administration. However, multiprobiotics (polyprobiotic Bifidobacterium animalis VKL and Bifidobacterium animalis VKB and composite probiotic Bifidobacterium animalis VKL, Bifidobacterium animalis VKB and Lactobacillus casei IMVB-7280) reduced the erosive and ulcerative lesions and the intensity of bleeding in rat GM when given within 14 days. It was shown that one of the mechanisms of antiulcer preventive effect of the multistrain probiotics is the restoration of pro/antioxidant balance in the GM under the stress action. The obtained results show the effectiveness of poly- and composite probiotics in the gastric ulcer prevention.

Regional variation in composition and antimicrobial activity of US propolis against Paenibacillus larvae and Ascosphaera apis.

Propolis is a substance derived from antimicrobial plant resins that honey bees use in the construction of their nests. Propolis use in the hive is an important component of honey bee social immunity and confers a number of positive physiological benefits to bees. The benefits that bees derive from resins are mostly due to their antimicrobial properties, but it is unknown how the diversity of antimicrobial activities among resins might impact bee health. In our previous work, we found that resins from different North American Populus spp. differed in their ability to inhibit in vitro growth of the bee bacterial pathogen Paenibacillus larvae. The goal of our current work was to characterize the antimicrobial activity of propolis from 12 climatically diverse regions across the US against the bee pathogens P. larvae and Ascosphaera apis and compare the metabolite profiles among those samples using LC-MS-based metabolomic methods. Samples differed greatly in their ability to inhibit both bacterial and fungal growth in vitro, but propolis from Nevada, Texas, and California displayed high activity against both pathogens. Interestingly, propolis from Georgia, New York, Louisiana, and Minnesota were active against A. apis, but not very active against P. larvae. Metabolomic analysis of regional propolis samples revealed that each sample was compositionally distinct, and LC-FTMS profiles from each sample contained a unique number of shared and exclusive peaks. Propolis from Aspen, CO, Tuscon, AZ, and Raleigh, NC, contained relatively large numbers of exclusive peaks, which may indicate that these samples originated from relatively unique botanical sources. This is the first study to characterize how the diversity of bee preferred resinous plants in the US may affect bee health, and could guide future studies on the therapeutic potential of propolis for bees.

[The changes of the motor function of the stomach and the colon under the action of the nanocrystalline cerium dioxide].

We investigated the effect of nanocrystalline cerium dioxide on parameters of spontaneous and stimulated motility of the stomach and colon in rats. It was found that administration of nanocrystalline cerium dioxide for 10 days increased the amplitude of contractions of stimulated motility in the stomach by 33.0 +/- 2.4% and the frequency of contractions of the colon by 80.3 +/- 7.5%. In this group, the introduction of carbachol also increased the frequency of the contractions by 274.0 +/- 22.9%, compared to the control group. The administration of nanocrystalline cerium dioxide increased the index of motor activity of spontaneous and stimulated motility of the stomach by 19.8 +/- 1.4 and 14.5 +/- 9.0%, respectively. In the colon, the motor activity increased by 14.3 +/- 1.1 and 11.1 +/- 0.8%, respectively. We also found that the nanocrystalline cerium dioxide rebuilt morphological condition of the mucous of the colon.

Ribonuclease activity of buckwheat plant (Fagopyrum esculentum) cultivars with different sensitivities to buckwheat burn virus.

Ribonucleases (RNases) are present in base-level amounts in intact plants, but this level is able to increase greatly under stress conditions. The possible cause for such an increase is protection against plant RNA-virus attack. Buckwheat burn virus (BBV) is a highly virulent pathogen that belongs to Rhabdoviridae family. In our study, we have analyzed the correlation between RNase activity and resistance of different buckwheat cultivars to BBV infection. Two cultivars, Kara-Dag and Roksolana, with different sensitivities to BBV have been used. Kara-Dag is a cultivar with medium sensitivity to virus and Roksolana is a tolerant cultivar. It has been shown that the base level of RNase activity in Roksolana cultivar was in most cases higher than the corresponding parameter in Kara-Dag cultivar. Both infected and uninfected plants of Roksolana cultivar demonstrated high RNase activity during two weeks. Whereas infected plants of Kara-Dag cultivar demonstrated unstable levels of RNase activity. Significant decline in RNase activity was detected on the 7th day post infection with subsequent gradual increase in RNase activity. Decline of the RNase activity during the first week could promote the virus replication and therefore more successful infection of upper leaves of plants. Unstable levels of RNase activity in infected buckwheat plants may be explained by insufficiency of virus-resistant mechanisms that determines the medium sensitivity of the cultivar to BBV. Thus, plants of buckwheat cultivar having less sensitivity to virus, displayed in general higher RNase activity.

[The effect of probiotic therapy on development of experimental obesity in rats caused by monosodium glutamate].

The effect of a mixture of probiotic strains (2:1:1 Lactobacillus casei IMVB-7280, Bifidobacterium animalis VKL, Bifidobacterium animalis VKB) on the development of experimental obesity in rats induced by neonatal administration of monosodium glutamate has been studied. It was shown that in rats of 4 months age, the injection of monosodium glutamate (4 mg/g) at 2, 4, 6, 8, 10 days after birth elicited abdominal obesity and metabolic syndrome. An intermittent administration of a probiotic mixture to rats treated with monosodium prevented the development of obesity. In the group of rats treated with probiotics, anthropometric parameters (weight and body length, Lee index, body mass index) did not differ from the level of intact rats. Visceral fat mass was decreased by probiotics by 38.5% (P < 0.05) compared to rats treated with water. Probiotics improved lipid metabolism: reduced the level of VLDL by 32.2% (P < 0,05), the level of LDL by 30.6% (P < 0.05), increased HDL by 25.7% (P <0,05) compared to obese control rats. Probiotic strains restored the secretion of adipocytes hormones (leptin and adiponectin) to the normal level of intact animals. The results show the effectiveness of probiotics for the prevention of obesity.

[Morphofunctional state of reproductive system of ageing male rats in case of using nanocerium].

The influence of nanocrystalline cerium dioxide (NCD, 1 and 100 mg/kg per os daily for 10 days) on morphofuctional state of reproductive system was investigated in ageing male rats. It has been established that activation of hormone-producing testicular Leydig's cells, as well as of secretory and proliferative processes in prostate, underlies the stimulating effect of NCD at a dose 1 mg/kg on hormonal function of testis and spermatogenesis of ageing male rats. NCD used at a dose 100 mg/kg had no significant effect on the assessed indices of morphofuctional state of reproductive system.

[Prophylactic effect of probiotic strains Bifidobacterium animalis VKL and VKB on stress-induced lesions in the gastric mucosa of rats].

It was investigated the effect of probiotic strains Bifidobacterium animalis VKL and VKB and their mixture on erosive and ulcerative lesions in the gastric mucosa (GM) of rats induced by water immersion restraint stress. It was found that separate prophylactic introduction for 14 days of Bifidobacterium animalis VKL or Bifidobacterium animalis VKB didn't protect the GM from erosive and ulcerative lesions induced by stress. Contrary prophylactic introduction of Bifidobacterium animalis VKL and VKB mixture has been effective in protecting the GM from the lesions. One of the mechanisms of the gastroprotection of these probiotic strains is prevention of mucus barrier from degradation, which was evident in decrease of free fucose and hexuronic acids content. These results confirm the expediency ofprobiotics use for the prevention of stress-induced lesions in the GM.

The effect of Lacto- and Bifidobacteria in monoculture on the vaginal microflora in norm and in cases of intravaginal staphylococcosis.

The influence of probiotic strains of Lactobacillus casei IMV B-7280, Lactobacillus acidophilus IMV B-7279, Bifidobacterium animalis VKL, Lactobacillus delbrueckii subsp. bulgaricus IMV B-7281 and Bifidobacterium animalis VKB, each strain separately, to the range of the urogenital tract microflora in physiological norm and in cases of experimental intravaginal staphylococcosis of mice induced by Staphylococcus aureus 8325-4. It was found that all these strains had different efficiency in Staphylococcus aureus 8325-4 growth suppression in the vagina of infected mice and affect the spectrum of microorganisms. Lactobacillus casei IMV B-7280 strain had effective antistaphylococcal effect and reduced the number of coliform bacteria and fungi in the vagina in cases of intravaginal staphylococcosis of mice. Lactobacillus casei IMV B-7280 strain is promising to create probiotic drugs effective in treating intravaginal staphylococcosis.

Antistaphylococcal action of lacto- and bifidobacteria and interleukn-2.

Antistaphylococcal action of Lactobacillus delbrueckii subsp. bulgaricus IMV B-7281, L. acidophilus LMV B-7279 and Bifidobacterium animalis VKB as well as interleukin-2 (IL-2) has been determined on the model of experimental staphylococcal infection in mice. It has been established that peroral administration of certain probiotic strains of lacto- and bifidobacteria or composition L. acidophilus LMV B-7279--L. delbrueckii subsp. bulgaricus IMV B-7281 with IL-2 to mice favored the reducing of terms of staphylococcus persistence in the kidneys of experimental mice. The studied schemes are promising for treatment of patients with surgical infection.

The effect of lacto- and bifidobacteria compositions on the vaginal microflora in cases of intravaginal staphylococcosis.

The effect of intravaginal injection of Lactobacillus casei IMV B-7280, Lactobacillus acidophilus IMV B-7279, Bifidobacterium animalis VKL and Bifidobacterium animalis VKB strains in various compositions on the range of microflora of the urogenital tract on the model of experimental intravaginal staphylococcosis of mice was determined. It was established that under the influence of various compositions of these strains changes in microflora spectrum occurred: the number of representatives of normoflora increased and the number of pathogenic microorganisms, including staphylococci, streptococci, coliform bacteria and fungi, significantly decreased. It was determined that strains of lacto- and bifidobacteria, that were studied, are prospective components of future probiotic drugs efficient in treating staphylococcosis.

Dose-dependent IFN-stimulating and immunomodulating properties of 6H-indolo[2,3-B] quinoxaline derivatives.

Two 6H-indoloquinoxaline derivatives were studied in different doses and schemes of application for their INFgamma-inducing potential and ability to effect functional activity of phagocytic cells. Tested compounds were shown to possess comparable or higher activity than reference drug Amixin in analogous doses. One indoloquinoxaline significantly elevated metabolic activity of macrophages and increased their potential for phagocytosis. Application of multiple treatments and higher doses allowed us to reveal differences between studied derivatives that were not obvious in previous in vivo experiment. Capacity of 6H-indoloquinoxalines to induce vast IFN amounts on in vivo level was demonstrated for the first time.