The effect of methyl jasmonate vapors on content of phenolic compounds in seedlings of common buckwheat (Fagopyrum esculentum Moench)

The effect of methyl jasmonate (MJ) vapors on content of phenolic compounds: free phenolic acids, total quercetin, and total phenolics in etiolated buckwheat seedlings were studied. The data presented show that low concentration of MJ (10-8 M) had no influence on trans-cinnamic acid (CA), but stimulated the accumulation of chlorogenic acid in hypocotyls and cotyledons of buckwheat seedlings. A moderate dose of MJ (10-6 M) did not change the level of chlorogenic acid in the hypocotyls and cotyledons, but CA synthesis was promoted in cotyledons, whereas in hypocotyls no significant effect was found. Highest concentration of MJ (10-4 M) caused small decline of CA in hypocotyls, but large stimulation of the acid production in cotyledons was noted. MJ had stimulatory effect on caffeic acid forming, but inhibited synthesis of vanillic acid in hypocotyls and cotyledons. Lowest concentration of MJ (10-8 M) elicited accumulation of quercetin glycosides in both studied tissues of buckwheat seedlings, however at higher doses (10-8 and 10-4 M) did not affect the flavonol level. The obtained results suggest that nonequivalent influence of methyl jasmonate on the phenolics composition can be a result of various mechanisms of MJ uptake, transforming and/or its translocation in buckwheat hypocotyls and cotyledons. Decline of anthocyanins level in buckwheat hypocotyls caused by MJ cannot be explained by enhanced accumulation of quercetin glycosides or free phenolic acids, but probably by synthesis of other unknown phenolic compounds

Częstość występowania mutacji czynnika V Leiden u pacjentów ze skłonnością do żylnej choroby zakrzepowo-zatorowej

Background. A tendency to venous thromboembolism is otherwise called hypercoagulability or thrombophilia. This disease can be acquired or have a genetic background, and may lead to pulmonary embolism. The basis for analysis and selection of treatment is genetic diagnosis, which detects the G1691A mutation in the factor V gene (factor V Leiden) – the best known congenital thrombophilia marker. Material and methods. The study was carried out in the years 2015-2017 on samples taken from patients (462 men and 1284 women) with a tendency to venous thromboembolism. Real-Time PCR was used to detect G1691A mutation in factor V gene. The analyses were performed in the Hematology Laboratory of the Center of Laboratory Medicine at the Medical University of Gdańsk. Results. Significant differences in the frequency of Leiden mutation were shown. This mutation predominated in men (25%), while in women G1691A mutation was detected with a 15% frequency (p=0.04). All possible genotypes were found among the subjects and the percentage of heterozygotes and homozygotes in both genders was similar. Conclusions. Congenital thrombophilia associated with G1691A mutation of factor V Leiden gene was found to be more common in men than in women. All possible genotypes were determined in the pool of test subjects. The mutation was most frequently detected in patients between 30 and 40 years of age, and rarely after 70 years of age.Wprowadzenie. Nadkrzepliwość (trombofilia) jest to skłonność do żylnej choroby zakrzepowo-zatorowej. Choroba ta może być nabyta lub mieć podłoże genetyczne i może prowadzić do zatorowości płucnej. Podstawą analizy i doboru leczenia jest diagnostyka genetyczna. Celem tej diagnostyki jest wykrycie mutacji G1691A w genie czynnika V (czynnik V Leiden) – najlepiej poznanego markera trombofilii wrodzonej. Materiał i metody. Badania zostały przeprowadzone w latach 2015-2017 na próbkach pobranych od pacjentów (462 mężczyzn i 1284 kobiet) ze skłonnością do żylnej choroby zakrzepowo-zatorowej. W celu wykrycia mutacji G1691A w genie czynnika V zastosowano metodę Real-Time PCR. Analizy zostały wykonane w Laboratorium Hematologii Uniwersyteckiego Centrum Medycyny Laboratoryjnej przy Gdańskim Uniwersytecie Medycznym. Wyniki. Wykazano istotne statystycznie różnice w częstości występowania mutacji Leiden. Mutację częściej występowała w grupie mężczyzn (25%), zaś u kobiet mutację G1691A wykrywano z częstością 15% (p=0,04). Wśród osób badanych stwierdzono wszystkie możliwe genotypy, a procentowy udział heterozygot i homozygot u obu płci był zbliżony. Wnioski. Trombofilia wrodzona związana z mutacją G1691A w genie czynnika V Leiden częściej występowała u mężczyzn niż kobiet. W puli osób poddanych testom oznaczono wszystkie możliwe genotypy. Mutację najczęściej wykrywano u pacjentów w przedziale wiekowym 30-40 lat, a najrzadziej po 70 roku życia

Wykrywalność antygenu D słaby w populacji potencjalnych biorców

Background. The strongest immunogen of the Rh system is the D antigen. It is found in several variants and categories, which makes it difficult to determine the correct RhD (Rh+) or RhD negative (Rh-) phenotype. Although only some of the varieties and types of this antigen are of clinical significance, it is important to determine the normal Rh phenotype in recipients and donors. The aim of this study was to determine the frequency of weak D antigen in a population of potential recipients. Material and methods. The study group consisted of selected blood recipients in whom weak expression of the D antigen or its antibody was detected. In order to estimate the expression of antigen D, the blood was analyzed in the laboratory of the Regional Center of Blood Donation and Blood Treatment in Lublin. Blood from 220 potential recipients (149 women and 71 men) were used in the conducted research. The clinical material from the Laboratory of Transfusion Serology at the Provincial Specialist Hospital in Biała Podlaska was also used. Results. The presence of a weak D was confirmed in 21 recipients. 4 cases of weak D were confirmed among recipients of blood transplant, while 17 cases among those who did not have blood transfusions. There were significant differences in the occurrence of the weak D in relation to the transfusion in both women (χ2 = 18.34 df = 2, p = 0.0001) and men (χ2 = 17.25). Conclusions. The correct determination of the RhD+ or RhD- phenotype is important for pregnant women who should be subjected to immunoprophylaxis of maternal-fetal conflict when a weak D is detected. In order to avoid post-transfusion complications among recipients, it is necessary to choose serologically and phenotypically crossed-matched blood components.Wprowadzenie. Najsilniejszym immunogenem układu Rh jest antygen D. Może on występować w kilku wariantach i odmianach, co stanowi trudność w ustaleniu prawidłowego fenotypu RhD dodatni (Rh+) lub RhD ujemny (Rh-). Chociaż tylko niektóre z odmian i warianty tego antygenu mają znaczenie kliniczne, to jednak istotne jest oznaczenie prawidłowego fenotypu Rh u biorców i dawców. Celem pracy było określenie częstości występowania antygenu D słaby w populacji potencjalnych biorców. Materiał i metody. Grupę badaną stanowili wyselekcjonowani biorcy krwi, u których wykryto słabą ekspresję antygenu D lub jego przeciwciała. W celu oceny wielkości ekspresji antygenu D, krew analizowano w pracowni Regionalnego Centrum Krwiodawstwa i Krwiolecznictwa w Lublinie. W przeprowadzonych badaniach przeanalizowano 220 potencjalnych biorców (149 kobiet i 71 mężczyzn). Wykorzystano też materiał kliniczny pochodzący z laboratorium Serologii Transfuzjologicznej przy Wojewódzkim Szpitalu Specjalistycznym w Białej Podlaskiej. Wyniki. O becność s łabego a ntygenu D p otwierdzono u 21 b iorców. B iorcom, k tórym przetaczano krew, w 4 przypadkach potwierdzono antygen D słaby, natomiast tym, którym nie przetaczano krwi, 17 przypadków. Wykazano istotne różnice w występowaniu antygenu D słaby w zależności od transfuzji zarówno w grupie kobiet (χ2 = 18,34 df = 2, p = 0,0001), jak i mężczyzn (χ2 = 17,25). Wnioski. Prawidłowe określenie fenotypu RhD+, RhD- jest istotne dla kobiet w ciąży, które w przypadku wykrycia antygenu D słaby powinny być poddawane immunoprofilaktyce konfliktu matczyno-płodowego. Chcąc uniknąć powikłań poprzetoczeniowych, biorcom należy dobierać zgodne serologicznie i fenotypowo składniki krwi

Effects of simultaneous use of methyl jasmonate with other plant hormones on the level of anthocyanins and biogenic amines in seedlings of common buckwheat (Fagopyrum esculentum Moench)

The aim of the study was to assess the impact of auxin (IAA), gibberellin (GA3) and cytokinin (kinetin), used solely and in combination with methyl jasmonate (MJ), on the accumulation of anthocyanins and biogenic amines in hypocotyls and cotyledons of common buckwheat (Fagopyrum esculentum Moench) seedlings. The obtained results indicate that accumulation of anthocyanins in buckwheat seedlings was dependent on the concentration of the phytohormone applied and the tissue studied. The combined use of MJ and IAA, GA3 or kinetin partly reversed the effect of strong inhibition of anthocyanin synthesis by MJ. IAA used solely decreased the level of anthocyanins in de-etiolated buckwheat cotyledons. IAA also caused a reduction of putrescine content, both in hypocotyls and cotyledons of buckwheat seedlings. MJ used alone caused high accumulation of 2-phenylethylamine (PEA) in buckwheat cotyledons and hypocotyls. The simultaneous application of MJ and IAA, GA3 or kinetin also stimulated PEA synthesis in buckwheat tissues, however this effect was significantly lower compared to the use of MJ only. A reverse significant correlation between PEA and anthocyanin contents occurred in buckwheat hypocotyls, but not in cotyledons. It was suggested that the deficiency of L-phenylalanine, a substrate for synthesis of 2-phenylethylamine, may be partly responsible for the decline in anthocyanin content in buckwheat hypocotyls under the influence of M

Wpływ traktowania propachizafopem na zawartość barwników i wzrost siewek kukurydzy cukrowej

Background. We examined the response of sweet maize seedlings (Zea mays var. saccharata L., cv. Złota Karłowa), as a non-target plant, to various doses of propaquizafop ((R)-2-[4-[(6-chloro-2-quinoxalinyl)oxy] phenoxy]-propionic acid 2-[[(1-methylethylidene)amino]oxy]ethyl ester) applied to the root zone or on shoots. Material and methods. The herbicide at concentrations of 0.56; 5.63 or 56.3 μM was used in a study on seedlings grown in hydroponic cultures in controlled light and temperature conditions. In the experiment the roots of maize seedlings were exposed to propaquizafop for 7 days. Also, seedling shoots were exposed by their immersing for 30 seconds in the investigated propaquizafop concentrations. We assessed the impact of propaquizafop on the seedlings elongation of primary roots and shoots. The content of anthocyanins in epicotyls and photosynthetic pigments (chlorophyll a, chlorophyll b, total carotenoids) in leaves were also measured. Results. A weaker effect on shoot growth was found when the roots of sweet maize seedlings were exposed to propaquizafop than was the case in the application of the herbicide to shoot. Application of the herbicide to the root zone also had little effect on the levels of chlorophylls and carotenoids, or on the ratios between them, in the leaves. The presumed responsibility for this effect is a weak uptake of the herbicide by the roots and / or its transport to the leaves of the maize seedlings. Conclusion. Propaquizafop has a smaller effect on sweet maize seedlings when applied to the root zone than it has after foliar exposition. A decline of anthocyanin content under the influence of low doses of propaquizafop suggests that it has a greater effect on the metabolism of phenylpropanoids than other inhibitors of acetyl coenzyme A carboxylase have.Badano reakcję siewek kukurydzy cukrowej (Zea mays var. Saccharata L., Złota Karłowa), jako rośliny niebędącej celem zwalczania, na różne dawki propachizafopu zastosowanego do strefy korzeniowej lub na pędy. Herbicyd stosowano w stężeniach 0,56; 5,63 lub 56,3 μM, na siewki rosnące hydroponicznie w kontrolowanych warunkach światła i temperatury. W eksperymentach siewki kukurydzy eksponowano na propaquizafop przez 7 dni. Aby ocenić wpływ propaquizafopu na siewki, mierzono wydłużenie korzenia głównego i pędu, a także zawartości antocyjanów w epikotylach i barwników fotosyntetycznych (chlorofile, karotenoidy) w liściach. Propaquizafop stosowany do strefy korzeniowej (pożywka) wywierał słabszy wpływ na wzrost pędów kukurydzy i niewielki wpływ na poziom chlorofilu i karotenoidów w liściach siewek kukurydzy niż zastosowany dolistnie. Efekty te są prawdopodobnie spowodowane niewielkim poborem przez korzenie i/lub translokacją herbicydu w siewce kukurydzy. Spadek zawartości antocyjanów pod wpływem małych dawek propaquizafopu sugeruje, że ma on większy wpływ na metabolizm fenylopropanoidów niż inne inhibitory karboksylazy acetylokoenzymu A

Methyl jasmonate vapors affect the composition and peroxidation of major fatty acids in common buckwheat seedlings

The effect of methyl jasmonate (MJ) vapors on the composition and peroxidation of major fatty acids in the organs of common buckwheat seedlings was investigated. The composition of fatty acids in the hypocotyl and cotyledons of seedlings changed significantly under exposure to MJ vapors in closed jars. Four-day exposure to MJ led to a significant reduction in the concentrations of stearic, linoleic, and linolenic acids in the hypocotyl, whereas oleic acid levels increased approximately 3.5-fold. A decrease in stearic acid levels and an increase in the content of linolenic acid were noted in cotyledons, whereas oleic acid levels decreased in roots. Seven-day exposure to MJ vapor caused a further reduction in stearic acid content and an increase in oleic acid and linoleic acid levels in the hypocotyl. At the same time, the linoleic acid content of roots and linolenic acid levels in cotyledons were doubled, but a 5-fold reduction in linolenic acid concentrations was observed in roots. Methyl jasmonate intensified fatty acid peroxidation in cotyledons after 4 and 7 days and in roots after 4 days of exposure. Peroxidation was inhibited in the hypocotyl and roots after 7 days. The noted changes in the composition and peroxidation of fatty acids are probably indicative of senescence in buckwheat seedlings under the influence of MJ. Senescence seems to proceed faster in cotyledons than in other organs of buckwheat seedling

Methyl jasmonate stimulates biosynthesis of 2-phenylethylamine, phenylacetic acid and 2-phenylethanol in seedlings of common buckwheat

Methyl jasmonate has a strong effect on secondary metabolizm in plants, by stimulating the biosynthesis a number of phenolic compounds and alkaloids. Common buckwheat (Fagopyrum esculentum Moench) is an important source of biologically active compounds. This research focuses on the detection and quantification of 2-phenylethylamine and its possible metabolites in the cotyledons, hypocotyl and roots of common buckwheat seedlings treated with methyl jasmonate. In cotyledons of buckwheat sprouts, only traces of 2-phenylethylamine were found, while in the hypocotyl and roots its concentration was about 150 and 1000-times higher, respectively. Treatment with methyl jasmonate resulted in a 4-fold increase of the 2-phenylethylamine level in the cotyledons of 7-day buckwheat seedlings, and an 11-fold and 5-fold increase in hypocotyl and roots, respectively. Methyl jasmonate treatment led also to about 4-fold increase of phenylacetic acid content in all examined seedling organs, but did not affect the 2-phenylethanol level in cotyledons, and slightly enhanced in hypocotyl and roots. It has been suggested that 2-phenylethylamine is a substrate for the biosynthesis of phenylacetic acid and 2-phenylethanol, as well as cinnamoyl 2-phenethylamide. In organs of buckwheat seedling treated with methyl jasmonate, higher amounts of aromatic amino acid transaminase mRNA were found. The enzyme can be involved in the synthesis of phenylpyruvic acid, but the presence of this compound could not be confirmed in any of the examined organs of common buckwheat seedlin

Exposure to Light of the Abaxial versus Adaxial Side of Detached <i>Kalanchoë blossfeldiana</i> Leaves Affects Anthocyanin Content and Composition Differently

The accumulation and composition of anthocyanins in leaves of Kalanchoë blossfeldiana, detached and kept for five days under natural light conditions, were investigated. The presence of fifteen derivatives of cyanidin, petunidin, and delphinidin was found. Changes in the content of each anthocyanin in the leaves before and after exposure to light on the abaxial (naturally upper) and adaxial (naturally lower) sides of the leaves were compared. When the adaxial side was exposed to light, the anthocyanin contents of the leaves did not change. In contrast, when the abaxial side of detached leaves was exposed to light, there was enhanced accumulation of delphinidin-rhamnoside-glucoside, cyanidin-rhamnoside-glucoside, cyanidin-glucoside-glucoside, and two unknown derivatives of petunidin and delphinidin. Application of methyl jasmonate (JA-Me) on the abaxial side exposed to light inhibited the accumulation of these anthocyanins. This effect could probably be due to the presence of these anthocyanins in the epidermal cells of K. blossfeldiana leaves and was visible in the microscopic view of its cross-section. These anthocyanins were directly exposed to JA-Me, leading to inhibition of their formation and/or accumulation. The lack of significant effects of JA-Me on anthocyanin mono- and tri-glycosides may indicate that they are mainly present in the mesophyll tissue of the leaf

Accumulation of Anthocyanins in Detached Leaves of <i>Kalanchoë blossfeldiana</i>: Relevance to the Effect of Methyl Jasmonate on This Process

Accumulation of anthocyanins in detached leaves and in excised stems of Kalanchoë blossfeldiana kept under natural light conditions in the presence or absence of methyl jasmonate (JA-Me) was investigated. When the abaxial surface of detached leaves was held lower than the adaxial surface (the normal or natural position) under natural light conditions, anthocyanins were not accumulated on the abaxial side of the leaves. In contrast, when the adaxial surface of detached leaves was held lower than the abaxial surface (inverted position), anthocyanins were highly accumulated on the abaxial side of the leaves. These phenomena were independent of the growth stage of K. blossfeldiana as well as photoperiod. Application of JA-Me in lanolin paste significantly inhibited anthocyanin accumulation induced on the abaxial side of detached leaves held in an inverted position in a dose-dependent manner. Anthocyanin accumulation in the excised stem in response to natural light was also significantly inhibited by JA-Me in lanolin paste. Possible mechanisms of anthocyanin accumulation on the abaxial side of detached K. blossfeldiana leaves held in an inverted position under natural light conditions and the inhibitory effect of JA-Me on this process are described. The accompanying changes in the content of primary metabolites and histological analyses were also described

Vasostatin increases oxygenation of B16-F10 melanoma tumors and raises therapeutic efficacy of cyclophosphamide

One of the preconditions of effective anticancer therapy is efficient transfer of the therapeutic agent (chemotherapeutic) to tumor cells. Fundamental barriers making drug delivery and action difficult include underoxygenation, elevated interstitial pressure, poor and abnormal tumor blood vascular network and acidic tumor milieu. In this study we aimed at developing an optimized scheme of administering a combination of an angiogenesis-inhibiting drug (vasostatin) and a chemotherapeutic (cyclophosphamide) in the therapeutic treatment of mice bearing experimental B16-F10 melanoma tumors. We report that the strongest tumor growth inhibition was observed in mice that received two, three or four vasostatin doses in combination with one injection of cyclophosphamide (i.e., V2 + CTX, V3 + CTX or V4 + CTX schemes). Double administration of vasostatin increases oxygenation of B16-F10 tumors. On the other hand, its five-fold administration lowers tumor oxygenation, breaks down tumor vascular network (increasing hypoxia) and leads in consequence to death of cancer cells and appearance of necrotic areas in the tumor. A decreased cyclophosphamide dose in combination with two doses of vasostatin (V2 + CTX scheme) inhibits tumor growth similarly to a larger dose of cyclophosphamide alone