Éva Sárdi

Micro-preparative OPLC — rapid isolation by transfusion and infusion-transfusion processes

A new OPLC procedure, infusion—transfusion OPLC, has been developed and compared with conventional transfusion OPLC. Spot and/or band deformation caused by the total wetness front (which results from pore filling) was reduced, as was the bubble effect in on-line detection. Both techniques were used for rapid micro-preparative OPLC isolation on analytical adsorbent layers. In-situ clean-up and separation were used to isolate trigonelline from Leuzea extract. Modeling of loading capacity for isolation of ascorbigen was accomplished by fully off-line OPLC. Under optimized conditions ascorbigen of high purity was isolated from cabbage extract by transfusion and infusion—transfusion OPLC.

Study of the defensive mechanism against drought in French bean (Phaseolus vulgaris L.) varieties

The response to drought of six green- and yellow-podded varieties of green bean was investigated in a phytotron. These varieties were also grown without irrigation in field experiments and analysis of leaf samples was carried out three times; before and under flowering and pod ripening, respectively. At first, the carotene and raffinose contents of leaves increase during mild drought (30/15°C). High temperature (35/25°C) combined with water deficiency resulted in a considerable decrease in leaf weight, chlorophyll b, maltose content, as well as the level of water-soluble antioxidants of the leaves, while the content of lipid-soluble antioxidants and raffinose content increased when compared with the control. The yellow-podded varieties of green beans responded to drought much more sensitively than the green-podded ones. The results suggested that a selection method based on the activity of antioxidants in the leaves can be used for testing the adaptability of numerous bean genotypes to drought.

Important bioactive molecules of erythrocytes in colorectal cancer patients after colectomy.

Formaldehyde (HCHO) and protoporphyrin, are in connection with redox homeostasis. Data show the importance of HCHO in proliferative as well as in apoptotic processes. Free protoporphyrin can be detected near the Zn-protoporphyrin in cancer and it has pro- and antioxidant forms depending on concentrations. The aim was to determine the amount of HCHO and protoporphyrin concentrations of erythrocytes in colorectal cancer after colectomy and to estimate redox homeostasis. Total 32 adult patients after 5-10 years of colectomy and 9 healthy volunteers were drawn into this study. Tumor markers, redox parameters, HbA1c, HCHO and protoporphyrin concentrations were measured. Erytrocyte HCHO was significantly lower in colectomysed patients, than in controls. Protoporphyrin concentration was low in patients, but in metastasis its concentration was significant. HbA1c correlated significantly with free radicals and decreased the antioxidant status of erythrocytes. HCHO and protoporphyrin concentrations of erythrocytes and the total scavenger capacity are very important indexes in cancer.

Relationship between peroxidase activity and the amount of fully N-methylated compounds in bean plants infected by Pseudomonas savastanoi pv. phaseolicola

Changes in the level of endogenous formaldehyde (HCHO), some N-methylated compounds (choline and trigonelline) and peroxidase activity were examined in the leaves of bean genotypes (Phaseolus vulgaris L.) with different disease-sensitivity during ontogenesis in the stressfree condition and after natural infection by Pseudomonas savastanoi pv. phaseolicola (until the appearance of lesions). HCHO, as its dimedone adduct, and fully N-methylated compounds were determined by overpressured layer chromatography (OPLC) in different developmental stages and in the infected leaves/leaf discs. Peroxidase activity was measured by a spectrophotometric method. HCHO level decreased with ageing of the primary leaf and accordingly in the leaves at different developmental stages, then increased again in both cases due to the demethylation and methylation processes. Concentration of choline and trigonelline as potential HCHO generators decreased considerably while peroxidase activity increased with ageing of the plants. Comparing the symptomless and the Pseudomonas infected leaf discs (with watersoaked lesions) we found a decrease in the level of HCHO, choline and trigonelline and there was detectable increase in the peroxidase activity in the infected leaf tissues. Our findings are in accordance with previously published results that peroxidases play an important role in oxidative demethylation processes. Our hypothesis is that the high level of HCHO in the old leaves can originate from methylated components as the result of peroxidase activity and this high level may lead to the old leaf being resistant to pathogen. This conclusion is supported by the fact that the leaves of susceptible bean genotypes became resistant to Pseudomonas while growing older.

How Can Cancer-Associated Anemia Be Moderated with Nutritional Factors and How Do Beta Vulgaris L. Ssp. Esculenta Var. Rubra Modify the Transmethylation Reaction in Erythrocytes in Cancerous Patients?

Genetic predisposition, unhealthy dietary or life habits and heavy social circumstances could result in the changes of redox homeostasis, which is very important for the equilibrium between tissue regeneration and apoptosis. When the balance is disturbed, cancer and/or necrosis may develop (Powis et al., 1997). Moderate dietary habits with natural bioactive agents, antioxidants, methyl donor molecules and metal elements can help restore the normal function of the organism, although the immoderate consumption of nutritive components is contraindicated. Long term antioxidant and/or antioxidant-related treatments as well as metal element owerflow can modify redox-homeostasis because these alimentary components effect signal transduction routes and compensatory effects of altered tissues can be observed (Vanherweghem et al., 1993, Blazovics et al., 2007a,b, Blazovics et al., 2008). Epidemiological, experimental and clinical investigations have shown that food supplements are not effective in cancer therapy because of inappropriate usage in cases of people suffering from cancers with low vitamin and trace element levels. Significant changes of total scavenger capacity, metal element concentrations, bounded HCHO and

Changes in carbohydrate content of potato calli during osmotic stress induced by mannitol.

Endogenous carbohydrate (fructose, glucose and sucrose) fractions were measured in calli of potato genotypes with different field tolerance to drought. Under in vitro stress conditions induced by 0.8 M mannitol, sucrose level of calli increased extremely in medium-tolerant (by 424.5%) and sensitive (by 302.7%) genotypes whilst the rate of increase was 12-18-times lower in the drought tolerant variety. Results suggest the applicability of sucrose as biochemical marker for distinguish drought tolerant genotypes from great population in callus culture.

Biochemical examination of the general defense system of plants by OPLC

Plant resistance breeding has, so far, been based on specific resistance genes which regulate hypersensitive reaction, resulting in fast tissue destruction. Hungarian researchers have, however, discovered the existence of the general defense system (gds), which regulates a completely different plant defense reaction. This manifests itself in tissue thickening through cell enlargement. We have studied the biochemical background of these different plant defense processes by OPLC, a forced-flow version of planar chromatography. We also exploited some basic well-known advantages of planar chromatography which made its application suitable and favorable in our experiments. As a result of analysis of time-dependent changes in carbohydrate compounds, we confirmed pathological and symptomatological observations that plants have both general and specific defense systems. These two systems have different roles and strategies in excluding pathogens, but together constitute the complete disease resistance of plants.

Ontogenesis-and Biotic Stress-Dependent Variability of Carbohydrate Content in Snap Bean (Phaseolus vulgaris L.)

Physiological examination of resistant and susceptible bean genotypes has shown that the concentration and quantitative ratios of carbohydrates measurable in leaf tissues depend on the age of leaves. During the phases of ontogenesis, the glucose and sucrose levels are the lowest in the primary leaves and highest in the youngest upper leaf. There is a continuous increase in the concentration of both carbohydrates from the oldest to the youngest leaves. The glucose/sucrose quantitative ratio decreases with ageing of the leaves until blooming. Our results indicate that the glucose concentration decreases considerably in the susceptible bean leaves after infection with the bean pathogen Pseudomonas savastanoi pv. phaseolicola. It has been proved that the glucose plays an important role in the formation of the bacterial extracellular polysaccharide (EPS) coat. Because there is a positive correlation between the age-dependent bacterial-resistance and the low sugar (especially glucose) content in older leaves of the originally susceptible bean plant, we think that in the old leaves there is not enough glucose for production of the EPS coat. Lacking EPS coat the bacterial and plant cell walls come in direct contact which permits the induction of hypersensitivity response characteristic of resistance

Effects of bioactive molecules of Beta vulgaris L. ssp. esculenta var. rubra on metastatic prostate cancer

Several reports are known about the effects of nutrition supplements in the improvement of quality of life of patients with tumor, however, the physiological background remains largely unknown. Table beet affects numerous biochemical reactions, enzymes and metabolic-synthesis. Methods: Natural table beet product come from commercial service was given twice 10g daily for 1 month for 24 patients (mean age 68±8 years) with hormone-resistant and metastatic prostate cancer treated with taxan chemotherapy, who report themselves fi rst, mean 3,6±2,8 years ago with their complains. 18 men’s data were amenable after treatment for evaluation. In addition to routine laboratory examination values of HbA1c, 9 cytokines and levels of 3 growth factors, the global parameters of redox-homeostasis, few elements of their metal-ions, Zn- and level of free protoporfi rin, trans-metilating processes before and 1 month after treatment were determined. Results: In most of the patients, favorable impact of beet was enforced and signifi cantly high levels of Zn- and free protoporfi rin decreased; furthermore, trans-metilating processes fastened. Conclusions: According to results, it seems that moderate and permanent consumption of table beet product affects the life expectancy of patients favorably; however, due to the increasing values of EGF, medical control is necessary for patients with prostate cancer treated by chemotherapy. Orv. Hetil., 2010, 37, 1495–1503.UNLABELLED Several reports are known about the effects of nutrition supplements in the improvement of quality of life of patients with tumor, however, the physiological background remains largely unknown. Table beet affects numerous biochemical reactions, enzymes and metabolic-synthesis. METHODS Natural table beet product come from commercial service was given twice 10 g daily for 1 month for 24 patients (mean age 68+/-8 years) with hormone-resistant and metastatic prostate cancer treated with taxan chemotherapy, who report themselves first, mean 3,6+/-2,8 years ago with their complains. 18 mens data were amenable after treatment for evaluation. In addition to routine laboratory examination values of HbA1c, 9 cytokines and levels of 3 growth factors, the global parameters of redox-homeostasis, few elements of their metal-ions, Zn- and level of free protoporfirin, trans-metilating processes before and 1 month after treatment were determined. RESULTS In most of the patients, favorable impact of beet was enforced and significantly high levels of Zn- and free protoporfirin decreased; furthermore, trans-metilating processes fastened. CONCLUSIONS According to results, it seems that moderate and permanent consumption of table beet product affects the life expectancy of patients favorably; however, due to the increasing values of EGF, medical control is necessary for patients with prostate cancer treated by chemotherapy.

Transmethylation and the redox homeostasis

INTRODUCTION Transmethylation modifies configuration and proceeds via formaldehyde. It has a significant role for example in epigenetic regulation. The whole methyl-pool can be evaluated by the measurement of bound formaldehyde. AIM The bound formaldehyde was measured in wheat, bean, beetroot, cabbage, broiler- and rabbit liver. The relationship between transmethylation and redox homeosthasis was studied in the liver of domestic animals, and in the rat model of fatty liver. METHOD The diet of rats was enriched with cholesterol, sunflower oil and cholic acid. The bound formaldehyde was determined by overpressured layer chromatography. The hydrogen-donating ability was measured with 1.1-diphenyl-2-picrylhydrazylt free radical using spectrophotometric measurement. RESULTS Beans had the most bound formaldehyde. The liver of broilers possessed significantly elevated hydrogen-donating ability and transmethylation ability. Rats with severe fatty liver had significantly less bound formaldehyde and the hydrogen-donating ability tendentiously decreased. CONCLUSIONS These results draw attention to the diet, especially in obesity and obesity-related diseases.