Betulae cortex, Betula pendula Roth., Betulaceae, comprise triterpene substances which are confirmed to posses very important pharmacological activities such as anti-inflammatory, anticancer and antiviral. In this study, extraction of triterpene substances from both, inner and external birch bark was carried out and after that qualitative analysis on betulin, betulinic acid, oleanolic acid and lupeol was performed by method of thin layer chromatography. By this separation method, applying system for development benzene-ethyl acetate-formic acid (36:12:5), is gained a good separation of examined triterpene substances from methanol extracts of inner and external birch bark as well as used standards. From obtained row triterpene mixtures, certain triterpene substances are isolated using method of dry column chromatography. To those substances infrared (IR) spectra were recorded and compared with IR spectra of adequate standards. The study encloses all obtained IR spectra and interpretations on the basis of which can be concluded that triterpene substances, betulin, betulin acid and lupeol isolated from external birch bark give identical characteristic signals and absorbance as referent standards. Method of dry column chromatography has resulted as simple, efficient, repeatable and economical for laboratory conditions. Beside this, a sufficient quantity of examined triterpene substances is also obtained for continuation of their further analytical analysis.

UNLABELLED The objective of this study was to evaluate cortisol in the saliva and serum of healthy persons and its daily fluctuations by using immunochemical method on a autoanalyzer. Biological samples: Serum from 14 healthy persons and saliva from 18 healthy persons were taken two times at 8 a.m. and at 4 p.m. Immunochemical assay: The principle of this method is the competitive binding of cortisol present in the analyzed sample and cortisol marked with peroxides on binding parts with specific antibodies. STATISTICAL ANALYSIS Student t-test. Cortisol in saliva in the morning: 21,2 +/- 16,2 nmol/l and in the afternoon 12,7 +/- 8,1 nmol/l. Cortisol in serum in the morning: 459, 6 +/- 235,2 nmol/l , and in the afternoon 340,5 +/- 207,5 nmol/l. The concentrations of cortisol in saliva are lower than in serum. Cortisol in the serum in the morning is about twenty times higher than cortisol in the saliva at the same time. Cortisol in the serum at afternoon is about twenty-seven times higher than cortisol in the saliva. Individual variabilities of cortisol in the saliva and serum were found during the day.

Aim of this study was to evaluate the biotransformation of simple phenols after ingestion of edible fruits and mixed food. It was analyzed hippuric acid in urine as biomarker of conjugation in the liver cells of glycine with aromatic phenolic acids such benzoic and salicylic acid from ingested food. Measurement of hippuric acid in urine samples of 10 healthy individuals: 5 female and 5 male with a mean age 51,5 years were recruited to participate in this study. Urine samples were collected for 24 hours. The additional meals 300 g of fruits: blueberry, cherry, raspberry, melon, blackberry and mixed food were given immediately before the 24 hr urine sampling. Otherwise, the meals given during 24 hr was a usually food. Biotransformation of phenols in edible fruits, that are together with liver glycins precursors of hippuric acid biosynthesis, was evaluated by direct spectrophotometric measurement of excreted hippuric acid in urine at 410 nm. It was established that the highest quantity of hippuric acid was after ingestion of 300 g of bilberry fruits (p< 0,003), and same quantity of cherries (p< 0,003). Concentration of excreted hippuric acid was twice higher after ingestion of these fruits in comparison with hippuric acid concentrations in urine after ingestion of common - mixed food. Quantity of biosynthesised hippuric acid was in direct correlation with the concentrations of its precursors, primarily phenol acids and other simple aromatic acids ingested with food.

Intracerebroventricular (icv) administration of betacytotoxic drug streptozotocin (STZ) produces long-term and progressive cognitive deficits in rats, as well as deficits in cerebral glucose and energy metabolism. These changes resemble those found in the brain of patients with sporadic Alzheimer's disease (sAD), and therefore, STZ-icv treated rats have been proposed as an experimental model of sAD. In this study the antioxidant capacity (AC), using manual oxygen radical absorbance capacity (ORAC) assay, was measured in the rat brain frontoparietal cortex (FC) and brainstem-cerebellum region (BS-CB) after administration of STZ and another betacytotoxic drug alloxan (AL). Region-specific differences of AC were found, which were more expressed when hydroxyl radical (ORAC(-OHo)) generator was used in the assay. AC against ORAC(-OHo) was significantly lower in BS-CB than in FC of the control rats. Furthermore, ORAC(-OHo) significantly decreased in BS-CB 3-months following the icv administration of AL, but significantly increased following the TG+AL combined treatment in comparison with the controls. However, 3-months following the icv treatment of AL combination with a different glucose transport inhbitor, 3-O-methyl-D-glucose, ORAC(-OHo) values in BS-CB and ORAC(-ROOo) values in FC were significantly decreased in comparison to the controls. Our results suggest that betacytotoxic-icv treatment alters antioxidant defense systems in the brain, which particularly regarding the STZ-icv treatment, could be a useful tool in search for possible new antioxidant treatments of the neurodegenerative disorders such as sAD.

The aim of this study was to investigate the antioxidant capacity (AC) in the lipophilic fraction of postmortem motorcortex (MC), nucleus caudatus (NC) and gyrus temporalis (GT) from controls (C) and Alzheimer's disease (AD) patients. The initial samples consisted of 50 human brain tissues of AD and C. AC of the different region of human brain were measured by using the fluorescent method of the oxygen radical absorbance capacity (ORAC). Peroxyl and hydroxyl radical generators were used in the analysis. All ORAC analysis were carried out on the Perkin-Elmer spectrofluorometer LS 55 with fluorescent filters, Ex: 485 nm; Em: 520 nm. Final results were calculated using the differences between area under the quenching curve of fluorescein (FL), blank and analyzed biological samples. AC against peroxyl radicals (ORAC-ROO degrees ) of lipophilic fraction in MC of AD was statistically significantly lower in comparison with MC of C (p < 0.008). No changes in the AC against hydroxyl radicals (ORAC- degrees OH) of lipophilic fraction of AD were found in comparison with C. Reduction of total protein in GT of AD (p < 0.03) was found. The results showed that in the MC of AD brain the balance between production of free radicals and the neutralization by a complex antioxidant system is disturbed. The manual fluorescent method for AC measurements proved to be sufficiently appropriate and sensitive for the AC measurements of lipophilic fraction of postmortem brain tissues from different patologic conditions.