The prolonged and uncontrolled use of antibiotics in treatment against many pathogens causes the multiple drug resistance. The drug resistance of Listeria monocytogenes F2365 has been evolved, which cause a major disease listeriosis. The drug dose limit against that pathogen was also increased for currently prescribed antibiotics and more often combinational therapy was preferred. Therefore, identification of an extensive novel drug target, unique and essential to the microorganism and subjected to its validation and drug development is imperative. Availability of the total proteome of L. monocytogenes F2365 enabled in silico identification of putative common drug targets and their subcellular localization by subtractive genomics approach. In the present work subtractive genomics approach is used to identify vaccine and drug targets of L. monocytogenes F2365 to speed up the rational drug and vaccine design. It has revealed that out of 2821 reference sequences of the pathogen, 744 represent essential proteins and among them 274 are human non-homolog proteins. Besides, all predicted human non-homologs were then analyzed by subcellular localization servers, in which 46 proteins were identified as surface exposed proteins and can be considered as potential drug and vaccine targets for the pathogen. The 3D structure of two human non-homolog putative drug targets, pantothenate kinase (LmPK) and holliday junction resolvase-like protein (LmHJR) of L. monocytogenes F2365 were generated by homology modeling program Easymodeller 4.0; a GUI version of modeller. Generated structures were also validated by several online servers. The overall stereochemical quality of the model was assessed by Ramachandran plot analysis that was provided by PROCHECK. ProQ, ERRAT, Pro-SA web and VERIFY 3D of SAVES programs were also used to compute several validation parameters during the evaluation of the model. This protein structure information is important in structure-based drug and vaccine design. This study provides information about putative drug targets of L. monocytogenes F2365 and 3D structures (LmPK and LmHJR), which emphasizes future perspective to design rational drugs and vaccines.

Medicinal plants have been intensively studied as a source of antitumor compounds. Due to the beneficial climate conditions Bulgarian herbs have high pharmacological potential. Currently, the antitumor effect of the Bulgarian medicinal plant Tribulus terrestris L. on human cancer cell lines is not studied. The main active compounds of the plant are the steroid saponins. The present study aims to analyze the effect on cell viability and apoptotic activity of total extract and saponin fraction of Bulgarian Tribulus terrestris L. on human breast cancer (MCF7) and normal (MCF10A) cell lines. Antitumor effect was established by МТТ cell viability assay and assessment of apoptotic potential was done through analysis of genomic integrity (DNA fragmentation assay) and analysis of morphological cell changes (Fluorescence microscopy). The results showed that total extract of the herb has a marked dose-dependent inhibitory effect on viability of MCF7 cells (half maximal inhibitory concentration is 15 μg/ml). Cell viability of MCF10A was moderately decreased without visible dose-dependent effect. The saponin fraction has increased inhibitory effect on breast cancer cells compared to total extract. Morphological changes and DNA fragmentation were observed as markers for early and late apoptosis predominantly in tumor cells after treatment. Apoptotic processes were intensified with the increase of treatment duration. The obtained results are the first showing selective antitumor activity of Bulgarian Tribulus terrestris L. on human cancer cells in vitro. Apoptotic processes are involved in the antitumor mechanisms induced by the herb. This results give directions for future investigations concerning detailed assessment of its pharmacological potential.

The essential oil of Salvia sclarea L., growing in Bulgaria, was analyzed by gas chromatography – mass spectrometry. A total of 52 different compounds were identified, representing 98.25% of total oil content. Linalyl acetate (56.88%) and linalool (20.75%) were determined as major essential oil constituents, followed by germacrene D (5.08%) and β-cariophyllene (3.41%). Antifungal activities of clary sage essential oil and major compounds linalyl acetate and linalool against 30 clinical isolates, belonging to species Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata and Candida parapsilosis were evaluated. Essential oil characterized with stronger anticandidial activity in comparison with pure compounds.

The genetic variability of four populations belonging to two introduced silkworm strains (Bombyx mori L.) of various origins has been studied using isoenzymic analysis of six enzyme systems. Nonspecific esterases, phosphoglucomutase, malate dehydrogenase, acid phosphatase, alkaline phosphatase and hexokinase from different tissue of larvae 5th instar have been analysed using PAGE. Polymorphism in six from a total of nine loci has been found. Inter- and intra-population differences have been ascertained expressed in different allele composition of the gene pool and different frequencies of alleles. A higher degree of inter-population variability has been reported on the acid phosphatase and a lower one – on the phosphoglucomutase.

One of the major challenges in modern synthetic polymer chemistry is to synthesize end defined polymers of different end functionality with predetermined uniform molecular weight. End functionalized polymers/copolymers basically in block and grafting form are having several potential applications in biomedical areas in the form of surface modifications, coatings, adhesives, and in order to increase the biocompatibility of polymeric blends. Among the existing controlled radical polymerization (CRP) methods for synthesis of these functional polymers, the atom transfer radical polymerization (ATRP) is one of the powerful techniques. The functional groups in these polymers can be easily introduced at the chain ends through functionalized ATRP initiators. A number of ATRP initiators have been developed in polymer science to develop defined polymer-protein bioconjugates. This critical review basically focuses on different types of ATRP initiators and their mechanisms used in the synthesis of polymer-protein bioconjugates.

The aim of this work was to study the production of extracellular lipase by solid-state fermentation with different olive cakes varieties including Mary, Shenghe and Yellow from isolated fungi using agro-industries waste such as rice straw, rice barn and wheat straw. The highest yields of enzyme were obtained in solid-state fermentation using rice straw as solid substrate in combination with 40% Mary olive cakes as inducer. The initial screening by using Plackett-Burman's design demonstrated that among the tested factors, lactose and ammonium sulfate of the medium significantly (p < 0.05) enhanced the lipase production. Further optimization of lipase production by isolated fungi in solid-state fermentation by applying response surface methodology was achieved, which revealed these as follows: 0.42 (% w/v) for lactose and 0.09 (% w/v) for ammonium sulfate. Also the enzyme kinetics parameters, biochemical properties, thermodynamic of thermal deactivation and deactivation rate constant of enzyme were determined.

Assessing the impact of mutagens for creating variations in crops like faba bean (Vicia faba L.) is an important criterion in the contemporary world where food insecurity and malnutrition is alarming at the doors of various nations. Impact of two chemical mutagens viz. hydrazine hydrate (HZ) and maleic hydrazide (MH) on the two varieties (NDF-1 and HB-405) of Vicia faba were analysed in terms of meiotic behavior and pollen sterility. Since there are not enough data about the effect of these mutagens on the chromosomal behaviors of Vicia faba, this study presents the role of hydrazine hydrate and maleic hydrazide as well as various types of chromosomal aberrations in crop improvement. The lower concentration of mutagens showed less pollen sterility compared to the higher concentrations. Manipulation of plant structural component to induce desirable alternations provides valuable material for the breeders and could be used favorably for increasing mutation rate and obtaining a desirable spectrum of mutation in faba beans based on preliminary studies of cell division.

A study on the trophic spectrum of three species of lacertid lizards (Lacerta agilis, Lacerta trilineata and Podarcis muralis) was carried out, based on 20 specimens collected in the period 1967-1973 in various localities in Bulgaria. The analyzed data showed that the insects (Insecta) are the most numerous and the most frequently met among the alimentary components of the total amount of food of the studied stomachs (except for Lacerta agilis, where spiders are slightly predominating). The non-insect components consisted spiders and isopods. The largest niche breadth was recorded in Lacerta trilineata (8.25), followed by Podarcis muralis (5.20) and Lacerta agilis (3.44). The niche overlap between the three species (pair-wise comparison) showed medium values and in our opinion there should not be any serious competition for food resources at the places with sympatric distribution.

Water-soluble and acid-soluble pectic polysaccharides with significant immunostimulating activity were isolated from alcohol-insoluble solids (AIS) of leek. Pectic polysaccharides characterized with high anhydrouronic acid content and common pectic sugars – L-arabinose, D-galactose, D-glucose, and L-rhamnose. It was accomplished that D-galactose content was the highest, which allowed the assumption that pectin sugar side chains were from galactan type. Enzyme modification of acid-extracted pectin was realized. Different pectin-degrading enzymes were used. The first modified pectic fraction, obtained after endopolygalacturonase action and alcohol precipitation contained rhamnogalacturonan I and had high D-galactose content. The second fraction obtained from the first, after β-galactosidase treatment characterized with low D-galactose content. Probably this was result from β-galactosidase hydrolysis of pectin side chains. It was proved by the action of β-galactosidase that galactan side chains were mainly composed of β-(1→4)-linked D-galactose. It was accomplished that rhamnogalacturonan I had higher immunostimulating activity than the initial pectic polysaccharide, which confirmed the statement that pectin hairy regions were responsible for the biological activity. The second fraction had lower D-galactose content and lower activity than the first. Probably galactan side chains were responsible for the immunostimulating activity.

The main purpose of this study was to investigate the effect of acute inflammation on total creatine kinase (CK) activity in dogs. In these animals, CK is an enzyme found predominantly in skeletal muscle and significantly elevated serum activity is largely associated with muscle damage. Plasma increases in dogs are associated with cell membrane leakage and will therefore be seen in any condition associated with muscular inflammation. The study was induced in 15 mongrel male dogs (n=9 in experimental group and n=6 in control group) at the age of two years and body weight 12-15 kg. The inflammation was reproduced by inoculation of 2 ml turpentine oil subcutaneously in lumbar region. The plasma activity of creatine kinase was evaluated at 0, 6, 24, 48, 72 hours after inoculation and on days 7, 14 and 21 by a kit from Hospitex Diagnostics. In the experimental group, the plasma concentrations of the CK-activity were increased at the 48th hour (97.48±6.92 U/L) and remained significantly higher (p<0.05) at the 72 hour (97.43±2.93 U/L) compared to the control group (77.08±5.27 U/L). The results of this study suggest that the evaluation of creatine kinase in dogs with experimentally induced acute inflammation has a limited diagnostic value. It was observed that the creatine kinase activity is slightly affected by the experimentally induced acute inflammation in dogs.