Polymers are used in several immunological applications, for example in designing new generation vaccines, implantation studies, therapeutics and for the development of animal models that are mimicking human pathological conditions. Polymers can function as attenuators to modulate host immune responses in tissue transplantation, as suppressors to inhibit immune responses against therapeutics or as an adjuvant in the activation of immune responses. Among them, polymers as adjuvants are highly promising and are being developed in the vaccination and autoimmunity fields. As an adjuvant, polymers can efficiently deliver antigens and have the ability to modulate immune responses toward an antigen. Adjuvant properties of the polymers are mainly dependent on their extrinsic and intrinsic properties such as polymer chemistry, format, charge and a fine balance between hydrophobicity and hydrophilicity. Polymers can also be easily coupled with an antigen/immuno-modulator/ligand either physically or chemically. Efficient delivery and protection against degradation of antigens in vivo are some of the advantages of these polymeric adjuvants. Present review focuses on various recent developments in polymeric adjuvants and responsible factors that affect their adjuvant properties.

Biomass accumulation and tropane alkaloids production by diploid and tetraploid hairy root cultures of Datura stramonium L. cultivated in stirred tank bioreactor at different aeration rates were investigated. The maximal growth for both hairy root cultures (ADB = 8.3 g/L and 6.8 g/L for diploid and tetraploid line, respectively) was achieved at aeration rate of 15.0 L/(L.h). The corresponding growth indexes were remarkably high (GIDW = 9.0 and 7.8 for diploid and tetraploid line, respectively) compared to the values, usually reported for other hairy root cultures. The optimal aeration rate for biomass accumulation was also optimal for alkaloids biosynthesis. According to our survey, the achieved maximal amounts of accumulated hyoscyamine (35.0 mg/L and 27.0 mg/L for diploid and tetraploid line) were the highest reported in the scientific literature for D. stramonium L. hairy roots. During the cultivation in stirred tank bioreactor, the hairy roots biosynthesized pharmaceutically important alkaloid scopolamine in minor concentrations. This is an important observation since scopolamine was not detected during submerged cultivation of these hairy root lines in other bioreactor types. However, the ploidy level was found to be the most important factor concerning scopolamine production by D. stramonium L. hairy roots cultures. The present work demonstrated the effect of ploidity levels on biomass accumulation and tropane alkaloids production by D. stramonium L. hairy roots cultivated in stirred tank bioreactor. This investigation show that the stirred tank bioreactor could be successfully applied for both maximal biomass accumulations, as well as for manipulation of tropane alkaloids production by diploid and tetraploid D. stramonium L. hairy root cultures.

Orange and lemon peels were used for obtaining pectic polysaccharides. Citrus peels were previously treated with 96% ethanol, and the obtained alcohol-insoluble solids (AIS) were subjected to a sequential extraction with hot distilled water and hot 0.5% HCl. Water- and acid-extracted orange (WEOP and AEOP) and lemon (WELP and AELP) pectins were obtained. Acid extraction gave higher yields of pectin than water extraction and lemon peels were richer in pectin. Comparative investigations were carried out with chromatographically purified commercial citrus pectin (CPCP). Chemical and physicochemical characterization of all pectins was accomplished. It was found that pectins were similar in anhydrouronic acid content (AUАC, 69-81%), but differed in their degree of methylesterification (DM, 55-81%). Generally water-extracted pectins were with higher DM. Both orange pectins were with higher DM and degree of acetylation (DA, 2%), in comparison with the corresponding lemon pectins. Water-extracted pectins were with higher degree of feruloylation (DF, 0.12-0.34%). To our knowledge this is the first report on the estimation of ester-linked ferulic acid in orange and lemon peel pectins. Pectic polysaccharides differed in molecular weight and homogeneity. WELP was with the highest molecular weight and homogeneity. The pectins contained D-galacturonic and D-glucuronic acids, L-arabinose, D-galactose, L-fucose, L-rhamnose and D-xylose. All investigated pectins showed immunostimulating activity by complement activation in the classical pathway at 1.25 and 2.5 mg/mL. Pectic polysaccharides were modified with endopolygalacturonase. Enzyme-modified CPCP and WEOP had higher anti-complementary activity than the corresponding initial pectins.

Glycosyltransferases are extracellular and cell-associated sucrase enzymes produced mainly by lactic acid bacteria Leuconostoc mesenteroides, oral Streptococcus species and also Lactobacillus species. According to the synthesized polymer (glucan or fructan) in the presence of sucrose, these enzymes are divided into two groups: glucosyltransferases (GTFs) and fructosyltransferases (FTFs). Only Streptococcus, Lactobacillus and Leuconostoc strains are known as producers of both GTFs and FTFs. The enzymes from Lactobacillus and Leuconostoc spp. are implicated in the synthesis of polymers and oligosaccharides (OS) important for human health because of their prebiotic properties and immunomodulating activity. In the present work, we studied the production of extracellular and cell-associated glycosyltransferases by Leuconostoc mesenteroides strain NRRL B-1149 during its growth on media containing glucose or fructose as a main carbon source. The enzyme activities, pH and biomass formation were measured and compared during the cultivation. We have shown that glucose and fructose have not an equal role for enzyme production. The highest extracellular activity was detected at the 4th hour during the cultivation of the strain in medium with fructose – 5.45 U/mg. When the strain was cultivated in medium with glucose, the maximum of extracellular enzyme activity was detected at the 5th hour of the cultivation but the measured activity was about 9 times lower compared to these, obtained after cultivation in fructose medium. The studied strain produced mainly extracellular glycosyltransferases in glucose or fructose medium, which were 92.4% and 97.1% of the total enzyme activity, respectively. In order to characterize the produced enzymes, cell-associated and extracellular enzymes were determined using SDS-PAGE and in situ Periodic Acid Schiff′s staining after incubation with 10% sucrose. When the investigated strain was grown in media with sucrose, glucose or fructose, several types of glycosyltransferases were detected – dextransucrase with molecular weight 180 kDa and two fructosyltransferases, corresponding to 120 kDa and 86 kDa molecular weights.

Proteins are involved in every aspects of life: structure, motion, catalysis, recognition and regulation. Today's highly sophisticated science of the modifications of proteins has ancient roots. The tailoring of proteins for food and medical uses precedes the beginning of what is called biochemistry. Chemical modification of proteins was pursued early in the twentieth century as an analytical procedure for side-chain amino acids. Later, methods were developed for specific inactivation of biologically active proteins and titration of their essential groups. Enzymatic modifications were mainly developed in the seventies when many more enzymes became economically available. Protein engineering has become a valuable tool for creating or improving proteins for practical use and has provided new insights into protein structure and function. The actual and potential use of milk proteins as food ingredients has been a popular topic for research over the past 40 years. With today's sophisticated analytical, biochemical and biological research tools, the presence of compounds with biological activity has been demonstrated. Improvements in separation techniques and enzyme technology have enabled efficient and economic isolation and modification of milk proteins, which has made possible their use as functional foods, dietary supplements, nutraceuticals and medical foods. In this review, some chemical and enzymatic modifications of milk proteins are described, with particular focus on their functional and biological properties.

The present study is a small part of a program for application the methods of passive and active phytomonitoring with herbaceous species, trees, mosses and lichens for assessment of the anthropogenic factor in urban conditions. Aesculus hippocastanum L. was studied as a possible biomonitor of air pollution with heavy metals and toxic elements in the town of Plovdiv (Bulgaria). Concentrations of Cd, Cr, Cu, Pb, V and U in leaf samples from urban areas with different anthropogenic impact were compared. Motor transport was found to be the major source of contaminants. It was found the significant contribution of some factors as urban gradient, canyon-street effect and wind rose in forming the urban air quality.

A rapid and accurate TLC-densitometry method for qualitative and quantitative determination of protopine has been developed. The best separation was achieved using a mobile phase chloroform – ethyl acetate – methanol – ammonium hydroxide (80:80:40:0.05, v/v/v/v). The results obtained by this method (CV% 3.4) corresponded well with those obtained by using a HPLC method. The reliability of the proposed method was proved through reproducibility test with alkaloid extracts from Fumaria spp.

The main goal of the present work is to study the effects of the new neonicotinoid insecticide „Actara 25 WG" on the intensity of expression of glycogen in the liver of common carp (Cyprinus carpio L.) by using PAS-reaction on cryosections. Common carp is an economically important fish species, which is widely used as a bioindicator for the health of freshwater basins since it could also survive at very contaminated sites. We have used 6.6 mg/L, 10 mg/L and 20 mg/L of the test chemical under laboratory conditions. The results demonstrated that the intensity of staining of the PAS-reaction is directly proportional to the increasing concentration of the insecticide. In addition, this indicates that the amount of glycogen in hepatocytes also increased. Conglomerates of accumulated glycogen in certain hepatocytes were found at the highest concentration of the insecticide. Therefore, we consider that under the influence of „Actara 25 WG" the process of glyconeogenesis in the liver of the studied fish accelerates.

This study was conducted to estimate the effect of microwave irradiation on fermentation pattern, metabolizable energy content and organic matter digestibility of tomato pomace using in vitro gas production technique. Tomato pomace samples were collected, pooled, ground and then exposed to 1000W microwave irradiation for 0, 2.5 and 5 minutes. The experimental samples were incubated in vitro with rumen liquor taken from three fistulated Iranian native cows at 2, 4, 6, 8, 12, 16, 24, 36, 48, 72 and 96 h. Cumulative gas production volume at early incubation times was significantly decreased by microwave treating time (P<0.01). Microwave irradiation could not significantly affect gas production parameters. Significant effects were not found on metabolizable energy content (7.82 - 8.02 MJ/ Kg DM) and organic matter digestibility (51.87% - 53.15%) of microwave treated tomato pomace. The results showed that microwave irradiation may be a useful method to decrease ruminal gas production (including methane), without any negative effects on nutritional value of tomato pomace.

This study was conducted to investigate the effects of dietary thyme (Thymus vulgaris), rosemary (Rosmarinus officinalis) and fenugreek (Trigonella foenum graecum) as feed additives on growth performance, proximate composition and ammonia excretion of European sea bass Dicentrarchus labrax. Four isonitrogenous (48% crude protein) and isocaloric (21 kj/g) diets were formulated to contain 0% (control) or 1% of thyme, rosemary or fenugreek. The thyme supplementation significantly increased protein efficiency ratio, fillet protein levels, protein and energy retentions (P<0.05). The medicinal herb additives did not change serum urea, uric acid, creatinine and ammonia excretion rate (P>0.05). The results indicate that dietary thyme improved the protein and energy retentions of sea bass.