Metabolomics Profiling of Tunisian Sonchus oleraceus L. Extracts and Their Antioxidant Activities

Sonchus oleraceus (L.) L. (Asteraceae) is an edible wild plant, known for its uses in traditional medicine. The aim of this study is to explore the phytochemical composition of the aerial parts (AP) and roots (R) of aqueous extracts of Sonchus oleraceus L. growing in Tunisia, using liquid chromatography-tandem mass spectrometry(LC/MS/MS), and determine the content of polyphenols and antioxidant activities. Results showed that aqueous extracts of AP and R contained, respectively, 195.25±33 μg/g and 118.66±14 μg/g gallic acid equivalent (GAE), and 52.58±7 μg/g and 3.2±0.3μg/g quercetin equivalent. AP and R extracts also contained tannins, 581.78±33 μg/g and 948.44±19 μg/g GAE. The AP extract in the 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activities, hydroxyl radical scavenging (OH−) and in cupric reducing antioxidant activity (CUPRAC) assays were respectively 0.325±0.036 mg/mL, 0.053±0.018 mg/mL, 0.696±0.031 mg/mL and 60.94±0.004 μMTE/g, while the R extract using the same assays showed, 0.209±0.052 mg/mL, 0.034±0.002 mg/mL, 0.444±0.014 mg/mL and 50.63±0.006 μM Trolox equivalent/g, respectively. A total of 68 compounds were tentatively identified by LC/MS/MS in both extracts in which quinic acid, pyrogallol, osthrutin, piperine, gentisic acid, fisetin, luteolin, caffeic acid, gingerol, were the most abundant in the LC/MS/MS spectrum. Many of these metabolites were found for the first time in Tunisian Sonchus oleraceus L. which may take account for the antioxidant activities exhibited by the plant.     

Parasitoids of chickpea leafminer Liriomyza cicerina (Diptera: Agromyzidae) and their parasitism rate on chickpea fields in North Tunisia

In recent years, as a consequence of climatic changes, the leafminer Liriomyza cicerina had emerged as a serious pest in chickpea crops causing considerable damage in Tunisia. Thus, the search of control methods is necessary. The aim of this research was to monitor and identify the naturally occurring parasitoids of the chickpea leafminer Liriomyza cicerina in insecticide-free chickpea-growing areas located in North-Western Tunisia. Chickpea leaves with mines were weekly sampled from Beja and Kef sites during 2016 and 2017 and kept to observe and count emerging leafminer and parasitoid adults in the laboratory. Results revealed that L. cicerina parasitoids complex species included three hymenopteran parasitoids namely Opius monilicornis (Fisher, 1962) (Braconidae); Diglyphus isaea (Walker, 1838) (Eulophidae) and Diaulinopsis arenaria (Erdös, 1951) (Eulophidae). These parasitoids were identified and recorded for the first time in Tunisia on L. cicerina with parasitism rate varied between 13% and 52.5%. The most common species was O. monilicornis. The correlation between infestation rate on chickpea plants and number of emerged three parasitoids was calculated revealing high significant and positive correlations. The diversity index (H′) was also calculated. The three parasitoids were found in two sites Beja and Kef; while the diversity differed during the developmental stage of chickpea crops.     

Comparative studies for serodiagnosis of haemorrhagic septicaemia in cattle sera

Haemorrhagic septicaemia caused by Pasteurella multocida is a major epizootic disease in cattle and buffaloes in developing countries with high morbidity and mortality rate. In the present study, a total of 88 P. multocida isolates were isolated from 256 nasopharyngeal swabs and lung tissues samples (34.4%) during the period from January, 2013 to March, 2014 from different governorates located in Egypt. Dead calves showed the highest percentage of P. multocida isolation followed by the emergency slaughtered calves, diseased calves then apparently healthy ones. These isolates were confirmed as P. multocida microscopically, biochemically by traditional tests and by API 20E commercial kit then by PCR. The percentages of positive serum samples using somatic antigen and micro-agglutination test at 1/1280 diluted serum were 10%, 54.49% and 0% in apparently healthy, diseased and emergency slaughtered samples, respectively whereas, the percentages using capsular antigen and indirect haemagglutination test were 40%, 60.89% and 60% in apparently healthy, diseased and emergency slaughtered samples, respectively. The ELISA showed the highest sensitivity for diagnosing P. multocida in apparently healthy, diseased and emergency slaughtered animals with percentages of 42%; 92.9% and 80%, respectively. The obtained results revealed that the ELISA using capsular antigen of P. multocida is a more sensitive and specific serological test for diagnosis of haemorrhagic septicaemia.     

Effect of heated naringenin on immunomodulatory properties and cellular antioxidant activity

Naringenin is one of the most popular flavonoids derived from citrus. It has been reported to be an effective anti-inflammatory compound. Citrus fruit may be used raw, cooked, stewed, or boiled. The present study was conducted to investigate the effect of thermal processes on naringenin in its immunomodulatory and cellular antioxidant activities. The effects of flavonoids on B and T cell proliferation were assessed on splenocytes stimulated or not with mitogens. However, their effects on cytotoxic T lymphocyte (CTL) and natural killer (NK) activities were assessed in splenocytes co-incubated with target cells. The amount of nitric oxide production and the lysosomal enzyme activity were evaluated in vitro on mouse peritoneal macrophages. Cellular antioxidant activity in splenocytes and macrophages was determined by measuring the fluorescence of the dichlorofluorescin (DCF). Our findings revealed that naringenin induces B cell proliferation and enhances NK activity. The highest concentration of native naringenin exhibits a significant proliferation of T cells, induces CTL activity, and inhibits cellular oxidation in macrophages. Conversely, it was observed that when heat-processed, naringenin improves the cellular antioxidant activity in splenocytes, increases the cytotoxic activity of NK cells, and suppresses the cytotoxicity of T cells. However, heat treatment maintains the anti-inflammatory potency of naringenin.     

QTLs mapping of morphological traits related to salt tolerance in <Emphasis Type="Italic">Medicago truncatula</Emphasis>

To understand the complex inheritance of tolerance to salt stress in Medicago truncatula, quantitative trait loci (QTLs) analysis was performed using a set of recombinant inbred lines (RILs) derived from a cross between the tolerant line Jemalong A17 and susceptible line F83005.5. The RILs and parental lines were grown in individual pots filled with sterilized sand in a greenhouse under 0 and 50 mM NaCl. Plants were harvested after a period of 60 days. Fourteen quantitative traits related to aerial and root growths were measured. Broad-sense heritability of measured traits ranged from 0.21 to 0.83 and from 0.05 to 0.62 in control and in salt-stressed conditions, respectively. Established correlations between measured traits are dependent on treatment effect. We identified and mapped 10 QTLs in control conditions and 19 in salt stress. No major QTL was identified indicating that tolerance to salt stress is governed by several genes with low effects. The QTLs detected under control and under salt-stressed conditions almost did not share the same map locations suggesting that the loci that are not stable across treatments reflect adaptation to this constraint. The maximum of QTLs was observed on the chromosome 8. The usefulness of these QTLs, identified in greenhouse conditions, for marker-assisted selection should therefore be evaluated under field conditions, and validated in other genetic backgrounds.     

Impact of TP53 codon 72 and MDM2 promoter 309 allelic dosage in a Moroccan population with hepatocellular carcinoma

Single-nucleotide polymorphisms (SNPs) in codon 72 of the TP53 gene (rs1042522) and in the promoter region of the MDM2 gene (SNP309; rs2279744) have been shown to play a role in the predisposition to many cancers. However, these findings were inconsistent in other tumor types, and ethnicity is suspected to be a critical factor influencing the effects of these SNPs on the cancer risk. The aim of the present study was to investigate whether these functional SNPs were associated with an enhanced risk of liver tumorigenesis in Moroccan patients. We have genotyped both polymorphisms in 96 patients with hepatocellular carcinoma (HCC) and 222 controls without HCC matched for age, gender and ethnicity by PCR-RFLP confirmed by sequencing. A joint effect between the MDM2 and TP53 polymorphisms and an increased risk of liver cancer was detected, with the odds ratio for the presence of both MDM2 309GG and TP53 72Pro/Pro genotypes being 10 (95% confidence interval 0.39-255.55). Interestingly, a significant increase in the HCC risk was observed when at least two deleterious alleles were present, indicating an allele dosage effect. There was no evidence for any association with early age of HCC onset when deleterious alleles of MDM2 SNP309 and TP53 Arg72Pro where present. Our findings suggest that the combination of TP53 72Pro and MDM2 309G polymorphisms enhance the risk of developing HCC. These results deserve further confirmation in other populations.     

Optimization of Spongiochloris sp. biomass production in the abattoir digestate

In the present study, the abattoir digestate was used as a culture medium for Spongiochloris sp. growth with added mineral components under optimized conditions in batch culture. Firstly, an Hadamard matrix was used to investigate the impact of certain influencing factors on the Spongiochloris sp. growth. Then, a fractional factorial design 2^7-4 was successfully employed to optimize the concentration of different added components to abattoir digestate for increased Spongiochloris sp. biomass production. The major influencing factors were NaHCO₃ and FeSO₄ at a level of 2000 mg/L and 5 mg/L, respectively. A high biomass production of 5.29 × 10⁶ cell/mL and an important content of chlorophyll a of 65.32 mg/L were obtained after 42 days of culture of Spongiochloris sp. on the defined abattoir medium at static conditions.     

The anti-inflammatory effect of paraoxonase 1 against oxidized lipids depends on its association with high density lipoproteins

AimThe aims of this study were to investigate whether purified PON1 can reduce the pro-inflammatory effect of oxidized phospholipids and whether the effect depended on its association with HDL.Main methodsLipid peroxidation was induced by copper ions and was measured using the conjugated diene method. Lysophosphatidylcholine (lyso-PC) formation was measured by HPLC with evaporative light scattering detection (ELSD) and ICAM-1 expression on Ea.hy926 endothelial cells was analyzed by flow cytometry.Key findingsPurified PON1 significantly inhibited copper-induced oxidation of LDL and HDL, causing a 60.5% and 77.7% decrease in conjugated diene formation, respectively. Incubating PON1 with oxLDL caused a significant increase in lyso-PC levels, while oxHDL caused a significant decrease. PON1 (12.5 to 50 μg/mL) had a pro-inflammatory effect in the presence of oxLDL, increasing ICAM-1 levels in Ea.hy926 cells by 33.0% and 40.6% (p < 0.001) respectively, and had an anti-inflammatory effect in the presence of oxHDL, causing a 3-fold reduction in ICAM-1 levels. PON1 also caused a significant decrease in TNFα? and purified lyso-PC-induced ICAM-1 expression. The results obtained with reconstituted HDL as well as LCAT and PAF-AH inhibitors suggested that the anti-inflammatory effect of PON1 against oxidized lipids is dependent on its association with HDL.SignificanceOur results clearly showed that PON1 is involved in the anti-inflammatory effect of HDL and that the effect appears to depend on its association with HDL.