First Report of Cabbage Head Rot Caused by Fusarium avenaceum in Poland.

Cabbage (Brassica oleracea L. var. capitata L.) is an important crop in Poland. Symptoms of a disease affecting cabbage were observed in 2012 and 2013 both in mid-August during the growing season and during storage in January and February. The disease affected about 30 to 40% of crops grown on ~9,000 ha over three locations: Charsznica in south Poland and Bedlno and Skierniewice in central Poland. Circular, watery lesions ranging from 10 to 60 mm in diameter on the surface of affected cabbage heads included whitish aerial mycelium that developed orange sporodochia in the center of each lesion. After 2 to 3 weeks, infection covered each entire cabbage head. A fungal pathogen was isolated from the orange lesions and from infected internal tissue. After sterilization of the excised tissue in 70% ethanol, the sections were each rinsed twice with sterilized water, dried on sterilized filter paper, and plated onto potato dextrose agar (PDA). Isolations consistently yielded morphologically homogeneous fungal colonies with abundant aerial mycelium that ranged from yellow to brownish yellow. The fungus produced pigmentation that changed the agar medium from dark yellow to brownish-burgundy. The mean colony growth was 66 mm after 7 days at 25°C. The fungus formed macroconidia, but microconidia and chlamydospores were not observed. Macroconidia were slender, slightly falcate, usually 3- to 5-septate, 44.7 to 60.7 × 3.7 to 5.5 µm, and formed in abundant orange sporodochia. On PDA, the isolates lost the ability to form sporodochia. Morphological and cultural features were typical of those of F. avenaceum (Fries) Saccardo (2). Koch's postulates were conducted to establish pathogenicity of each of four of the isolates on cabbage heads of the cv. Jaguar F1 (Bejo Seeds, Poland). The outer leaf of each head was inoculated with an 8-mm-diameter PDA plug colonized by the appropriate isolate (four cabbage heads/isolate), and the heads stored in a growth chamber at 25°C. After 5 to 7 days, lesions similar to those observed on naturally infested cabbage were observed on all the inoculated cabbage leaves. Four cabbage heads treated similarly with water as a control treatment remained symptomless. The experiment was repeated. DNA extracted from two of the four isolates was subjected to a PCR assay with primers ITS5 and ITS4 (4) for species identification based on the ITS1 and ITS2 sequences of ribosomal DNA (rDNA). The two sequences differed by 1 bp in the ITS2 region and had 100% identity with ITS sequences of F. avenaceum Accession Nos. AY147283 and AY147285 in GenBank. The sequences were deposited in GenBank as KM189440 and KM189441. Descriptions of fusarium head rot of cabbage in the United States (1) and Canada (3) were consistent with these observations in Poland. To our knowledge, this is the first report of F. avenaceum causing head rot of cabbage in Poland and in Europe. References: (1) H. R. Dillard and A. C. Cobb. Phytopathology 96:30. 2006. (2) J. F. Leslie and B. A. Summerell. Page 132 in: The Fusarium Laboratory Manual, Blackwell Publishing, Hoboken, NJ, 2006. (3) R. D. Peters et al. HortSci. 42:737. 2007. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Mesoionic oxatriazoles (MOTA): NO-donating characteristics and pharmacology.

Biological role of nitric oxide (NO), functioning of isoforms of NO synthetases (NOS) and pharmacology of principle NO-donors were reviewed. NO donating characteristics and pharmacology of 23 mesoionic oxatriazoles (MOTA) were compared with those of 5-morpholinosydnonimine (SIN-1), S-nitroso-N-acetylpenicillamine (SNAP), sodium nitroprusside (NaNP) and glyceryl trinitrate (GTN). It is concluded that in vitro NO donating profile of MOTA hardly can be used as a predicting measure for their pharmacological activities either in vitro or in vivo. If anything, fast NO releasers seem to be stronger vasorelaxants than MOTA with slow NO releasing properties. Still, among representatives of this last category of MOTA one may find efficient antithrombotic and thrombolytic agents. For instance, MOTA 5-oxides were more potent thrombolytics than SIN-1, SNAP or NaNP. Also MOTA with potent anti-platelet action in vitro seem to be potent relaxants of tracheal strips. In summary, by manipulating the chemical structures of MOTA one may obtain relative selectivity towards vasorelaxant, anti-platelet, thrombolytic or tracheorelaxant properties. Thus different categories of MOTA might be designed with a hope of achieving hypotensive, antithrombotic, thrombolytic or anti-asthmatic drugs.

Influence of bezafibrate on total antioxidant activity of serum and deformability of blood cells in hypercholesterolaemic patients.

Fourteen patients with hypercholesterolaemia were treated with bezafibrate (600 mg/day) for a month. Before and after the treatment some biochemical and physical parameters were investigated. Plasma cholesterol levels, elevated before treatment, significantly decreased after one month (p<0.001). Deformability of red blood cells increased significantly in this time (p<0.05). Total antioxidant status of serum decreased statistically significantly (p<0.01). It is concluded that the mechanism of action of bezafibrate is not connected with resistance to oxygen free radicals since the drug has no antioxidant activity. A main mechanism of action of bezafibrate seems to be lowering the cholesterol level. An increase in deformability of red blood cells may make some contribution to this mechanism.

Nitric oxide donation and nitrite assays in the presence of thiols and albumin as determined by Griess' and Werringloer's methods.

Nitric oxide (NO) or nitrite (NO2-) were assayed using the Werringloer's method or the Griess' method, respectively, in the presence or absence of various thiols, amino acids, or albumin. This has been done because both methods are used to determine the generation of endogenous NO from L-arginine or exogenous NO from drugs in vivo, paying little attention to biological constituents which may affect results of these assays. Albumin, reduced glutathione (GSH), cysteine and N-acetylcysteine, but not other amino acids lowered the amount of NO2- as detected by Griess' method no matter whether sodium nitrite or 3-morpholinosydnonimine (SIN-1) were used as a source of NO2-. This happened probably because at low pH of the reaction mixture the corresponding nitrosothiols were formed and thus NO2- was not accessible for detection. However, this phenomenon was not seen when instead of SIN-1 another NO donor--S-nitroso-N-acetylpenicillamine (SNAP) was used. SNAP is a nitrosothiol itself and physiological low molecular thiols (e.g. GSH or cysteine) displaced NO from SNAP. An increase in the amount of released NO was detectable by both Werringloer's and Griess' methods. Only the presence of 700 microns of albumin steadily suppressed the detection of NO or NO2- no matter what was the source of these species. It is concluded that low molecular thiols and albumin may differently influence the detection of both NO and NO2- which derive from various NO donors or sodium nitrite.

Fused 2-thiohydantoin derivatives: evaluation as potential antioxidants.

A series of fused 5,5-diphenyl and 5-arylidene-2-thiohydantoin derivatives were examined for their oxygen free radical inhibitory and radical scavenging properties (RSC) using both an enzymic and non-enzymic biological generators of free radicals. Non-enzymic lipid peroxidation (OH. radicals) was assayed as the amount of malondialdehyde (MDA) that had been formed during incubation of boiled rat liver microsomes in the presence of ascorbic acid and ferric ions. Superoxide anions (.O2- anion radicals) were generated enzymatically in the xanthine-xanthine oxidase system. Among the 20 investigated compounds only four fused arylidene 2-thiohydantoin derivatives showed weak antioxidant activity scavenging OH. radicals. There is a relationship between the electrophilic OH. radical scavenging properties (RSC) of the tested molecules and the value of their HOMO energy derived from semiempirical MO calculations.

Evaluation of mercaptoalkyl derivatives of imidazolidine-4-one as potential antioxidants and free radical scavengers.

Mercaptoalkyl derivatives of 5,5-diphenyl- and 5-arylidene hydantoins were synthesized as the result of hydrolytic cleavage of bicyclic imidazothiazoles, -thiazines, and -thiazepines. The title compounds were evaluated as potential antioxidants and free radical scavengers using the enzymic generation of O2- and non-enzymic lipid peroxidation tests. Examined compounds were inactive in both applied tests, where reference substances (flavonoids, nitric oxide donors and Ticlopidine) inhibited non-enzymic lipid peroxidation and scavenged superoxide anions.

Bioactivity of flavonoids.

Flavonoids are benzo-gamma-pyrone derivatives of plant origin. They possess wide spectrum of biological activity. From the therapeutical point of view the most important are their antioxidant properties. These are the result of high propensity to electron transfer, ferrous ions chelating activity and direct scavenging of reactive oxygen species. Flavonoids inhibit enormous number of enzymes. From the pharmacological point of view inhibition of cyclooxygenase and lipoxygenases as well as scavenging of superoxide anions seem to be essential. Flavonoids are antiinflammatory agents as the result of diminished formation of proinflammatory mediators (prostaglandins, leukotrienes, reactive oxygen species, nitric oxide). They are also antithrombotic owing to their ability to scavenge superoxide anions. These anions are strong inhibitors of prostacyclin production. Removal of superoxide anions by flavonoids facilitates antiaggregatory PGI2 formation. Superoxide anions generate proagregatory isoprostanes. The antiaggregatory effect of flavonoids may be due to the limitation of formation of isoprostanes. Empirical use of flavonoids as drugs acquired recently scientific confirmation.

Thrombolytic action of ticlopidine: possible mechanisms.

Ticlopidine (Ticlide), an anti-platelet drug with a broad scope of clinical applications, is claimed to be an antagonist of adenosine diphosphate on platelet receptors. In vitro this antagonism cannot be demonstrated. Ex vivo it is detectable many hours after oral administration of the drug, perhaps subsequently to its biotransformation to an unknown metabolite. Here, we report for the first time that in patients with peripheral arterial disease and in cats with extracorporal circulation ticlopidine evokes instantaneous thrombolytic or fibrinolytic effects which are not associated with inhibition of platelet aggregation. Shortening of euglobulin clot lysis time and increase in plasma levels of tissue plasminogen activator were observed 1-2 h after oral ingestion of ticlopidine at a single dose of 500 mg. In cats ticlopidine produced instantaneous anti-thrombotic and thrombolytic effects at doses of 0.3-1 mg/kg and 10-15 mg/kg i.v., respectively. Thrombolysis by ticlopidine (10 mg/kg i.v.) was comparable to that by prostacyclin at a dose of 0.3 microgram/kg i.v. Ticlopidine at a concentration of 100 microM increased endothelial thromboresistance in vitro. The drug did not inhibit the activity of cyclooxygenase-1 or 12-lipoxygenase while it inhibited lipid autooxidation (IC50 = 18 microM) in rat liver microsomes. Our data point to a possibility that the therapeutic efficacy of ticlopidine might be associated not only with its delayed anti-platelet effects but also with its immediate thrombolytic action which is likely to be mediated by endothelial prostacyclin and tissue plasminogen activator rather than by platelet mechanisms.

Scavenging of reactive oxygen species as the mechanism of drug action.

Reactive oxygen species (ROS) are generated when oxygen is supplied in excess and/or its reduction is insufficient. The best explored ROS are superoxide anions, hydroxyl radicals and hydrogen peroxide. The first two are free radicals. ROS are harmful for the living cells and are implicated in a variety of pathological processes and diseases. Drugs used in the treatment of these states are either stimulators of endogenous defense mechanisms against ROS or inhibitors of ROS formation. Six groups of anti-ROS substances have been described in this paper. 1) Antioxidant substances used in substitutive therapy such as enzymes (e.g. superoxide dismutase), substances containing thiol groups and vitamins (A, E, P, C). 2) Chelating agents (e.g. desferoxamine), which lower the level of prooxidative transition metal ions. 3) Inhibitors of superoxide ions generation by stimulated cells or xanthine oxidase. Such mechanism of action was described for xanthine oxidase inhibitor-allopurinol. 4) Superoxide scavengers. Many known drugs were investigated for this activity, but the best documentation was presented for flavonoids. 5) Substances which eliminate hydrogen peroxide, mainly glutathione and its precursors. 6) Scavengers of hydroxyl radicals. Studies of the above activity were conducted mainly using an unspecific method--estimation of malondialdehyde generated during the action of hydroxyl radicals on lipids or on desoxyribose. Inhibition of malondialdehyde formation was described for many drugs of plant and synthetic origin.

Interactions between reactive oxygen species and sulfhydryl groups of cysteine, acetylcysteine and glutathione.

Oxidative potencies of hydrogen peroxide, superoxide anions or hydroxyl radicals towards sulfhydryl groups of cysteine, acetylcysteine or glutathione were studied. Acetylcysteine was the least susceptible to oxidation by reactive oxygen species. The most vigorous oxidant of sulfhydryl groups was hydrogen peroxide.

In vitro generation and decomposition of S-nitrosothiols from direct and indirect nitric oxide donors.

Nitric oxide which was released in aqueous solutions (> or = 10 microM) of direct NO-donors such as 3-morpholinesydnonimine (SIN-1) and S-nitroso-N-acetyl-penicillamine (SNAP) consumed avidly sulfhydryl groups of N-acetylcysteine > cysteine > glutathione. In case of SIN-1 generation of nitrites run in parallel to disappearance of sulfhydryl groups of N-acetylcysteine and glutathione, however, for a pair of SIN-1 and cysteine the rate of formation of nitrites was much slower than the rate of consumption of sulfhydryl groups. We infer that kinetics of formation and breakdown of S-nitrosothiols varies depending on the type of a thiol which reacts with a NO-donor. Indirect NO-donors such as glyceryl trinitrate (GTN), molsidomine (MSD) or sodium nitroprusside (NaNP) at concentrations < 100 microM did not consume sulfhydryl groups of cysteine unless pretreated with the xanthine/xanthine oxidase system. We suppose that in this last case superoxide anions react with nitric oxide to form peroxynitrites with a higher potency than nitric oxide itself to destroy sulfhydryl groups. We conclude that out of three studied thiols N-acetylcysteine is the best substrate for the formation of S-nitrosothiols, while S-nitrosocysteine is the slowest releaser of nitric oxide. Moreover, unlike SIN-1 and SNAP, NaNP is not a direct NO-donor but behaves rather like GTN. Minute amounts of nitric oxide released either from NaNP or GTN gain from superoxide anions an amplification as SH-scavengers.

[Evaluation of fear and analysis of personality structure of women at risk for premature and after term labor]. / Ocena leku oraz analiza struktury osobowosci kobiet zagrozonych porodem przedwczesnym oraz porodem po terminie.

Psychological studies were performed in 57 pregnant women at risk for premature labour and 36 at risk for after term labour. The level of fear was evaluated using the Spielberger STAI questionnaire. The structure of fear and restlessness was studied using the R.B. Cattell's five factor Self-estimation Sheet. The analysis of personality structure was performed on the basis of results of the MMPI-WISKAD test. In the patients in both studied groups, groups an increased level of situational and persistent fear was found. Women at risk for premature labour are characterized by higher level of fear of neurotic character resulting from disharmonious personality. In women delivered after term the fear is of reactive character. Women at risk for after term labour are characterized by a higher degree of personality integration and more effective ability to control fear. Women at risk for premature labour are characterized by disharmonious personality and simultaneously they use more primitive protective mechanisms. Women delivered after term are characterized by higher degree of personality integration and more effective ability to control fear.

Clinical and morphological analysis of endometriosis in women of the Lublin Macroregion between 1987 and 1992.

Histopathological examinations performed between 1987 and 1992 at the Department of Pathology of the Lublin Medical Academy were analysed statistically. Endometriosis was found in 885 cases, i.e. 13.3% of all gynecological cases. An increased frequency as well as a shift in the peak age of incidence to the third decade, were found.

Synthesis and biological evaluation of derivatives of N-[4-substituted-1-piperazinylalkyl]-1-(butyl,aryl)-2,5- dimethylpyrrole-3,4-dicarboxymide (Part II).

A series of new N-[4-substituted-1-piperazinyl-alkyl]-3,4-pyrroledicarboxyimides 7 have been prepared by reaction of N-haloalkylimide derivatives 6 with corresponding N-monosubstituted piperazines. Compounds 7 tested in preliminary pharmacological investigation produced a general depressive action on the central nervous system.

Nitric oxide donors as generators and scavengers of superoxide anions.

Three classical direct nitric oxide (NO) donors, 3-morpholine-sydnonimine (SIN-1), S-nitroso-N-acetyl-d,l-penicillamine (SNAP) and sodium nitroprusside (NaNP) as well as two indirect NO donors, molsidomine (MSL) and glyceryl trinitrate (GTN) were studied for their potencies to generate O2-, to scavenge O2-, to consume molecular oxygen and to inhibit lipid oxidation. Out of five NO donors only those which were spontaneous releasers of NO at physiological pH were also scavengers of O2- which has been generated by xanthine:xanthine oxidase system (SIN-1 IC50 19 microM, SNAP IC50 416 microM) and inhibitors of the Fe3+ and ascorbate stimulated oxidation of rat liver lipids (SIN-1 IC50 76 microM, SNAP IC50 12 microM). Only SIN-1 at high concentrations of 300-5000 microM generated O2- as detected by a SOD inhibitable reduction of nitroblue tetrazolium. None of the in vitro studied activities were exerted by NaNP, MLS and GTN.

Studies on antioxidative activity of some C-glycosylflavones.

Ten flavonoid C-glycosyl derivatives: orientin (1), isoorientin (2), vitexin (3), isovitexin (4), isovitexin 7,2"-di-O-glucoside (5), isovitexin 7-O-galactoside-2"-O-glucoside (6), two different 6,8-di-C-hexosylapigenins (7, 8), and two different 6-C-hexosyl-8-C-pentosylapigenins (9, 10) have been either produced from flavonoid fractions from Adonis vernalis L. (1, 2) and Crataegus species (3, 4), or isolated from Stellaria media (L.) Vill. (5-10) to study their antioxidative properties. These were found only for two compounds: orientin (1) and isoorientin (2).

The application of ultrasounds for detection of scavenging of superoxide anions by drugs.

In aqueous solutions ultrasounds are known to generate oxygen free radicals. Here we report that ferricytochrome c and nitroblue tetrazolium when sonificated in 1% ethanolic buffered solutions are reduced predominantly in a SOD-inhibitable manner. Aqueous solutions of adrenaline undergo oxidation when exposed to ultrasounds. Therefore, it seems that in the presence of ethanol or adrenaline ultrasounds generate substantial amounts of superoxide anions along with other free radicals. We have tried to adapt ultrasound technique for detection of scavenging of superoxide anions by a metabolite of molsidomine, SIN-1 and flavonoids. In the presence of SIN-1 reduction of indicators by superoxide anions generated by ultrasounds was prevented. In the case of quercetin we failed to detect this property because ultrasounds were found to transform native flavonoids into oxidized derivatives.

Ultrasound-induced oxidation of flavonoids.

Ultrasound-induced oxidation of some flavonoids was detected by UV spectral analysis. From among twenty seven compounds investigated only flavonol aglycones and their 7-glycosides, but not 3-glycosides, flavones, catechins and flavanones were found to be oxidized with opening of the ring C. The products of quercetin oxidation were identical with those obtained during slow aerial oxidation of the compound in neutral solution. The oxidation products were deprived of the oxido-reductive properties of native flavonoids. Neither they reduced cytochrome c, nor inhibited lipid oxidation nor scavenged superoxide anions.

The influence of sulfonated bioflavonoids on enzymatic oxidation of arachidonic acid and on non-enzymatic lipid oxidation.

Sulfonic acids of quercetin and morin as well as their ferrous and cupric complexes were synthetized and investigated. Sulfonic derivatives of quercetin were much weaker inhibitors of soybean lipoxygenase than quercetin itself. Morin and its derivatives were inactive. Antioxidant properties of quercetin derivatives were in the same range as for quercetin. Most of the investigated compounds stimulate cyclooxygenase when 100 microM of arachidonic acid is used as a substrate. Ferrous complex of quercetin 5'-sulfonic acid was an inhibitor of this enzyme.

Screening of the influence of flavonoids on lipoxygenase and cyclooxygenase activity, as well as on nonenzymic lipid oxidation.

Thirty nine flavonoids, isolated from plants, were tested in respect of their influence on soybean lipoxygenase activity, cyclooxygenase activity and inhibition of ascorbic acid-stimulated malonaldehyde formation in liver lipids. Almost all of the tested compounds were antioxidants and stimulated cyclooxygenase when arachidonic acid was used as a substrate at a concentration of 100 microM. Eleven flavonoids were inhibitors of soybean lipoxygenase. A good correlation between the chemical structure and the tested activity was observed. The most active compounds in all tests were luteolin, 6-hydroxyluteolin, nepetin, quercetagetin, patuletin and myricetin.