The study of arbuscular mycorrhizal symbiosis (AMS) - the mutualist relationship between the most of land plants and evolutionary old fungal group Glomeromycota - is becoming a prestigious topic. The prevalence of and extent of physiological action of AMS on plants is very interesting for the plant biology itself, but its importance grows, notably in time of global climate change, frequent soil degradation and ascending exhaustion of mineral fertilizer reserves. Although the flows in AMS of some minerals, like of phosphorus was enlightened, carbon exchange between the symbionts is still poorly understood. In this experimental work, I utilized the boom of molecular and bioinformatic methods in the quest for completely unexplained carbon flows. The organisms used include barrel medic (Medicago truncatula), the model legume for symbiotic relationships, biotic, and abiotic stresses; Rhizophagus irregularis, the widely used fungus for such experimental studies of AMS; and Sinorhizobium meliloti, the nodulating nitrogen-fixing bacterium compatible with the barrel medic. Two variants - mycorrhizal (M+) and non-mycorrhizal (NM) plants were subjected to several levels of analysis. I have checked the variants, did the measurements of phosphorus and nitrogen contents, as well as I probed the plants with...

Presented diploma thesis was focused on study of biologically active compounds, especially some sugars in cereal products. In experimental part the total of 29 different cereal materials, food industry waste products and natural complex matrixes (microbial polysaccharide, honey, beer) were used. As a part of this work optimization of analytical methods suitable for analysis of simple sugars, oligo- and polysaccharides was done. In cereal samples reduced and neutral sugars were analyzed spectrophotometrically and individual sugars were detected by chromatography. For HPLC/RI analysis optimal mobile phase composition and chromatography conditions were proposed. For mono- a oligosaccharides C18-NH2 sorbent, mixture acetonitril:water 75:25 as mobile phase and flow 1 ml/min were verified as suitable separation parameters. Thin layer chromatography of mono- and oligosaccharides was optimized too. Introduced chromatography and spectrophotometry methods were then applied to analysis of cereal samples. As a model sugar natural microbial polysaccharide pullulan was used to analytical method testing. In cereal products and food matrixes total neutral and reduced sugars as well as products of their acid and enzyme hydrolysis were measured. Detailed analysis of some glycoside composition was tested too. As the most usable method for both qualitative and quantitative analysis of cereal sugars HPLC/RI method was found. To detailed identification of malto-oligosacharides tandem LC/MS/MS technique using derivatization by 1-phenyl-3-methyl-5-pyrazolon was tested too.

Presented diploma thesis was focused on study of phenolic substances in various cereal products. Flours, brans and germs from various cereals, flakes and extruded cereals were chosen as samples. Phenolic substances were analyzed by spectrophotometry, RP-HPLC/UV-VIS and RP-HPLC/PDA/ESI-MS. Studies of saccharide content which were analyzed by spectrophotometry and HPLC/RI was incorporated to this thesis. Differences in the content of phenolic substances released from glycosides after acid hydrolysis of samples and differences in saccharide content determined after acid and enzymatic hydrolysis of samples were investigated. Simple sensory analysis of extruded cereal products of different flavour (cocoa, honey, chocolate) was carried out too. In analyzed samples the highest content of total phenolics and total flavonoids was measured in wheat germs. Especially in wheat germs and also in buckwheat and oat flakes, maize brans and cereals products Teddy (cereal balls with cocoa) and Ferda (cereal grains with honey) high levels of individual catechins, phenolic acids and flavonoids were determined too. The quantity of phenolic substances increased for several times in samples processed by acid hydrolysis in comparison to non-hydrolyzed samples. The high content of total saccharides, reduced saccharides and individual mono- and disaccharides was found mainly in extruded products. Concentration of monosaccharides increased in acid hydrolyzed samples in spite of disaccharides concentration that was higher only in some of hydrolyzed samples. In several cases final disaccharide concentrations were lower in hydrolyzed samples than in non-hydrolyzed ones.

Presented diploma thesis is focused on analysis of selected biologically active substances in different kinds of natural and peeled rice. In rice samples phenolic compounds, antioxidant activity, content of saccharides and vitamin C were analysed. Active substances and saccharides were analysed by UV-VIS spectrophotometry and RP-HPLC-UV/VIS. Content of phenolic compounds and saccharides was measured before and after acid hydrolysis. The content of these compounds increased several times because of release of active substances from glycosides. As a part of diploma thesis basic sensory analysis was performed and consumer questionnaires were evaluated. The highest level of antioxidant activity was detected in Indian Wild rice. The highest content of total polyphenols and flavonoids was detected in Red Rice Natural before acid hydrolysis. After acid hydrolysis the highest content was detected in Indian Wild rice. The highest content of individual flavonoids, phenolic acids and catechins was detected in natural rice Indian Wild, Rice Three colours and in two samples of rice parboiled. The highest content of total and reducing saccharides was found in rice Indian Wild before hydrolysis, while after hydrolysis the content was very similar in all rice samples. The amount of individual monosaccharides after acid hydrolysis increased, while total content of disaccharides decreased. The highest value of simple sugars was detected in rice parboiled and Indian Wild rice. The content of vitamin C was detected only in four rice kinds - in two samples of rice parboiled, Rice Three colours and rice Indian Wild.

Present diploma thesis describes the study of bioactive compounds in the cereals. The work is focused on possible fortification of cereal products in order to increase nutritional value of final products which is reduced during the food processing and storage. Cereal grains are rich in several phenolic compounds, vitamins and minerals that reduce the risk of chronic diseases. The aim of this study was the preparation and analysis of model fortified cereals using lyophilized fruit originating from Czech Republic. A comparative analysis of content of bioactive substances in commercial and fortified products for children was performed too. The experimental part deals with a content of individual flavonoids by RP-HPLC/UV/VIS method, spectrophotometric determination of total and reducing saccharides and analysis of mono- and disaccharides by the HPLC/RI method as well. Differences in the content of individual flavonoids and saccharides after the application of acid hydrolysis of samples were investigated. The content of these compounds increased in most products because of their release from glycosidic forms. Presence of phenolics was confirmed in all analyzed cereals. Addition of 10% of lyophilized fruit/vegetable preparative (carrot, berries, apples) caused several fold increase of phenolics content in model mixtures. In commercially available cereal products there are relatively high differences in total phenolics content, proportionally to fruit component amount. The work included the spectrophotometric analysis of total polyphenols and flavonoids in a model physiological environment (artificial stomach juice) in which the hydrolysis of bound phenolic compounds occurred. The results of this study show that addition of lyophilized fruit seems to be a good way to increase the nutritional value and maintain the content of bioactive compounds in cereal products.

The aim of this master’s thesis is perform basic analyses of samples ketchups. The theoretical part describes the basic characteristics of tomato, tomatoes processing and production of puree and ketchup. Then there are described analytical methods which were used for analysing the selected parameters in ketchups. The experimental part deals with the determination of pH, total solids, organic acids, saccharides, macroelements and lycopene in ketchups. Citrate and acetate were analysed by IC. The amount of acetate in ketchups was about 5 times greater than the amount of citrate. Glucose, fructose and sucrose were analysed by HPLC. Glucose and fructose were determined in all samples but sucrose not. The ICP-OES method was used to determine four elements. Magnesium was the least represented element and sodium was the most represented element in the analysis. The amount of sodium was recalculated on the amount of salt. Lycopene was analysed by UV-VIS spectrometry. Its amount in ketchups is influenced by the variety of used tomatoes and plenty of other factors.

In this thesis was determinated selected chemical and nutritional parameters in the wild elderberry and 17 cultivated varieties of elderberry. The fruits was provided by the Research and Breeding Institute of Pomology Holovousy Ltd. The theoretical part deals describes elderberry, chemical composition of elderberry, especially biological active substances and its using not only food industry. Closer attention has been focused on effect of saccharides, anthocyanins and methods of their determination, specially determination by HPLC. The experimental part describes the various methods of determination of selected chemical and nutritional parameters. Total dry matter, content of total polyphenolic compounds, monomeric anthocyanin pigment and antioxidant activity was defined in all varieties of elderberry fruits. For some varieties was defined total soluble dry matter, content of organic acids and saccharides by various methods. In the end was compared the values from different methods varieties.

Polysaccharides are complex biopolymers that play essential roles in the biological systems including energy storage, structural support, lubrication, and signal transduction. Despite their importance, the synthesis of polysaccharides has proven to be very challenging due to the presence of multiple hydroxyl groups and difficulty in controlling the stereochemical outcome of glycosylation reactions. As a conventional chemical method to synthesize polysaccharides, ring-opening polymerization of anhydrosugars enables the synthesis of stereoregular α-(1→6)-linked polysaccharides, but is less effective in preparing polysaccharides with other linkages. Enzymatic polymerizations have also been explored, however, these methods typically require expensive monomers, and suffer from a narrow scope of enzymes and small scale of reactions. The limited approaches to polysaccharides have inspired chemists to synthesize polysaccharide mimetics with achiral linkages that can be constructed efficiently. Poly-amido-saccharides (PASs) are a new type of saccharide polymers in which the O-glycosidic linkages in natural polysaccharides are replaced with (1→2)-amide linkages. With saccharide moieties inter-connected by amide bonds, PASs exhibit characteristics of both polysaccharides and polypeptides, such as possessing pyranose-backbones and lots of hydroxyl groups, and adopting a left-handed helical conformation. However, due to lack of sufficient terminal saccharide residues, previously synthesized glucose and galactose PASs display weak interactions with carbohydrate binding lectins and receptors, limiting their applications in biomedical and pharmaceutical fields. Herein, the design and synthesis of PASs with novel structures and properties is described. By pre-installing the stereochemistry in the monomer, Altrose PASs (Alt-PASs) with β-(1→2)-amide linkages are prepared via ring-opening polymerization of an altrose-based β-lactam followed by debenzylation. Circular dichroism shows that Alt-PASs adopt a right-handed helical conformation in aqueous solution. Via the polymerization of disaccharide-based β-lactams, two PASs with either 4-O-α-D-glucose branches (Mal-PASs) or 6-O-β-D-glucose branches (Gen-PASs) are obtained. Biological studies reveal that Mal-PASs are multivalent ligands to lectin Concanavalin A, while Gen-PASs activate RAW 264.7 macrophage cells by enhancing the secretion of TNF-α and NO. The anionic ring-opening polymerization of sugar-based β-lactams is a useful method to synthesize well-defined polysaccharide mimetics, and this method expands the current repertory of approaches available to complex saccharide polymers with biological activities. / 2021-02-15T00:00:00Z

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Thesis (M.S.); Submitted to Iowa State Univ., Ames, IA (US); 19 Dec 2004. / Published through the Information Bridge: DOE Scientific and Technical Information. "IS-T 2488" Tugba Kalem. US Department of Energy 12/19/2004. Report is also available in paper and microfiche from NTIS.