Distribution of phenolic compounds and antioxidant activity between young and old leaves of Carthamus tinctorius L. and their induction by salt stress

In the present work, we investigated the effect of salt stress on the distribution of safflower (Carthamus tinctorius L.) antioxidant system in relation to leaf age. The study was carried out under growth chamber conditions using seedlings of three cultivars which were subjected to 0 and 50 mM NaCl for 3 weeks. Leaf growth, water content, lipid peroxidation, and phenolic compound (total polyphenols, total flavonoids, and proanthocyanidins) concentration were measured at two leaf stages (young and old leaves). Leaf growth was affected by salinity only in Kairouan cultivar that also showed a significant decrease in old leaf water content. By contrast, Gabes and Tazarka cultivars maintained their old leaf water content constant and showed a reduction in that of young leaves. This could be attributed to a higher aptitude of the latter two cultivars to use absorbed sodium and chloride for osmotic adjustment in old leaves, keeping potassium for specific functions. Salt-induced lipid peroxidation was observed only in old leaves, whereas the accumulation of the major phenolic compounds under saline conditions was higher in young leaves, except in Gabes cultivar where no significant difference was found between the two leaf stages. A significant variability was also found between the three cultivars. The better behavior of salt-challenged leaves of Gabes and Tazarka cultivars compared to that of Kairouan cultivar may be related to their higher water content and the accumulation of polyphenols, in particular flavonoids that were shown to be efficiently involved in the restriction of salt-induced oxidative damages.     

Transgene copy number-dependent rescue of murine beta-globin knockout mice carrying a 183 kb human beta-globin BAC genomic fragment

We report the generation and characterisation of the first transgenic mice exclusively expressing normal human beta-globin ((hu)beta-globin) from a 183 kb genomic fragment. Four independent lines were generated, each containing 2-6 copies of the (hu)beta-globin locus at a single integration site. Steady state levels of (hu)beta-globin protein were dependent on transgene copy number, but independent of the site of integration. Hemizygosity for the transgene on a heterozygous knockout background ((hu)beta(+/0), (mu)beta(th-3/+)) complemented fully the hematological abnormalities associated with the heterozygous knockout mutation in all four lines. Importantly, the rescue of the embryonic lethal phenotype that is characteristic of homozygosity for the knockout mutation was also demonstrated in two transgenic lines that were homozygous for two copies of the (hu)beta-globin locus, and in one transgenic line, which was hemizygous for six copies of the (hu)beta-globin locus. Our results illustrate the importance of transgene copy number determination and of the hemizygosity/homozygosity status in phenotypic complementation studies of transgenic mice containing large heterologous transgenes. Transgenic mouse colonies with 100% (hu)beta-globin production from the intact (hu)beta-globin locus have been established and will be invaluable in comparative and gene therapy studies with mouse models containing specific beta-thalassemia mutations in the (hu)beta-globin locus.     

Feeding ecology of the vulnerable aoudad (Ammotragus lervia) in north-western Sahara

The feeding ecology of the aoudad (Ammotragus lervia) was investigated for the first time in north-western Sahara, Djebel Antar (Bechar province, Algeria), from autumn 2015 to summer 2016. Microhistological analyses of faeces revealed an annual diet composed of 23 identified taxa belonging to 16 plant families. The highest species diversity was recorded in spring and summer (23 species), despite a marked consumption of two species: Vachellia tortilis (17.7%) and Avena sterilis (14.0%); diet diversity was lower in autumn and winter (16 species), including mainly Teucrium polium (14.7%, 21.0%) and Gymnocarpos decander (19.7%, 10.0%). The main plant parts consumed during these seasons were stems (77.7%, 65.3%), while leaves and inflorescences were mostly consumed during spring and summer (54.7%, 52.3%). Forbs dominated the aoudad's diet, with 46.3% average relative abundance, including mainly T. polium, Limoniastrum feei, and Chrysanthemum macrocarpum. Woody plants including mainly V. tortilis and G. decander accounted for 33.3% (50.0% in autumn), and grasses including A. sterilis and H. murinum for 20.4% (32.0% in summer). Based on this diet, A. lervia can be classified as a “generalist mixed-feeder.”     

Design,Synthesis, Cytotoxic Screening and Molecular Docking Studies of Novel Hybrid Thiosemicarbazone Derivatives as Anticancer Agents

Thiosemicarbazones have been the focus of scientists owing to their broad clinical anticancer range. Herein, A Series of new thiosemicarbazone derivatives 5 – 9 were synthesized and confirmed through the use of different spectroscopic techniques along with elemental analysis. The in vitro cytotoxic activity of compounds 5 – 9 against MCF-7 and A549 cell lines and normal breast cells were assessed. Several compounds were found to be active. The most active compound 7 caused MCF-7 cell cycle arrest at G1/ S phases; and induced apoptosis at the pre-G1 phase. The apoptosis-inducing activity of compound 7 was proofed by the elevation of caspase 3/7 activity and also by up-regulation of the expression of Bax and p53 proteins together with the down-regulation of the expression of the Bcl-2 protein. It also had a strong inhibitory effect topoisomerase IIβ enzyme. Molecular Docking study revealed that the synthesized compounds had good docking scores compared to the standard drug Etoposide towards the topoisomerase IIβ protein (3QX3). Overall, these findings confirmed that the new thiosemicarbazone derivatives could aid in the development of promising cancer drug candidates.     

Evaluation and identification of reliable reference genes for toxicological study in Caenorhabditis elegans

To identify reliable reference genes for toxicological studies, 16 commonly-used reference genes were selected as candidates to evaluate their expression stabilities under experimental conditions in Caenorhabditis elegans. Sixteen candidates were composed of 12 protein-coding genes and 4 non-coding RNAs, they were act-2, ama-1, arp-6, cdc-42, csq-1, eif-3.C, idhg-1, mdh, pmp-3, rbd-1, tba-1, Y45F10D.4, 18S rRNA, Ce234, U18, and U6. Larval stage 1 synchronized hermaphrodites were exposed to benzo-α-pyrene (BαP), chlorpyrifos, diazinon, gossypol, zinc oxide nanoparticles, and the vehicle control DMSO for 30 h, respectively. Expression stabilities of candidate genes were analyzed using 4 independent evaluating approaches (BestKeeper, the delta Ct approach, geNorm, and NormFinder) followed by a comprehensive method. Results showed that there were slight differences in ranking order between evaluation methods due to their different assumptions and computations. The results also showed that responses of candidate genes to different chemicals were distinct, 18S rRNA was the best for BαP and chlorpyrifos, tba-1 was the most stable gene for diazinon and gossypol treatments, while pmp-3 was more stable for zinc oxide exposure. Additionally, results demonstrated that combinations of multiple genes were more reliable than individual gene, suggesting selecting two or more candidates as reference genes may generate more reliable results for toxicological studies.     

Different antioxidant responses to salt stress in two different provenances of <Emphasis Type="Italic">Carthamus tinctorius</Emphasis> L.

Seedlings of two Tunisian Carthamus tinctorius L. provenances (Kairouan and Tazarka) differing in salt sensitivity were hydroponically grown at 0 and 50 mM NaCl over 21 days. Leaves of Kairouan (salt-sensitive) showed a 48% restriction in their growth at 50 mM NaCl although they accumulated less sodium than those of Tazarka (less salt-sensitive) that maintained an unchanged growth. Salt treatment induced oxidative stress in C. tinctorius and the effect was more pronounced in the leaves of the more salt sensitive provenance, Kairouan. Both provenances exhibited a stimulation of antioxidant enzyme activities with higher catalase (CAT) and superoxide dismutase (SOD) activities in Tazarka and higher peroxidase (POD) activity in Kairouan. But, it seems that antioxidant activities were more correlated with polyphenol content. Actually, leaves of Tazarka experienced higher polyphenol and antioxidant activity than Kairouan at 50 mM NaCl. Hence, moderate salinity (3 g NaCl L⁻¹) enhanced bioactive molecule yield in the less salt sensitive provenance, Tazarka. In addition, C. tinctorius was found rich in ascorbic acid, but the moderate salt stress enhanced its production only in the sensitive provenance.     

Validated spectrofluorimetric and resonance Rayleigh scattering methods for determining naftidrofuryl in varied pharmaceutical samples based on its interaction with erythrosin B

Naftidrofuryl is a vasodilator medication used for treating cerebral and peripheral vascular diseases. In this study, two spectroscopical techniques, spectrofluorimetric and resonance Rayleigh scattering (RRS), were utilized to quantify naftidrofuryl in its pharmaceutical samples. The developed methodologies in this study rely on a facile process of forming an association complex between erythrosine B reagent and naftidrofuryl under acidic conditions. The fluorimetric assay is based on the ability of naftidrofuryl to quench and decrease the native fluorescence intensity of the reagent when measured at ${\lambda }_{\mathit{\text{emis}}.}$ = 550 nm ( ${\lambda }_{\mathit{\text{excit}}.}$ = 526 nm). Under similar reaction conditions, the RRS method relies on the observed amplification in the RRS spectrum of the reagent at a wavelength of 577 nm following its interaction with naftidrofuryl. The methods exhibited linearity within the ranges 0.2–1.6 μg/ml (r² = 0.999) and 0.1–1.4 μg/ml (r² = 0.9994), with limit of quantitation values of 0.146 and 0.099 μg/ml, and limit of detection values of 0.048 and 0.032 μg/ml, for the fluorometric and the RRS methods, respectively. Moreover, the quenching between the dye and naftidrofuryl was studied using Stern–Volmer analysis, and the methodologies were experimentally optimized and validated. Additionally, acceptable recoveries were achieved when the procedures were applied to determine naftidrofuryl in pharmaceutical samples.     

Distribution,population dynamics and damage of Hessian fly,Mayetiola destructor (Diptera: Cecidomyiidae) in North Tunisia

A three years survey and monitoring studies (2013–2014–2015) were carried out through 4 regions of north Tunisia in order to follow the evolution of the distribution, the frequency of occurrence and damage caused by the Hessian fly Mayetiola destructor (Say) to bread wheat (Triticum aestivum L.) and durum wheat (Triticum durum Desf). Moreover, the effectiveness of resistance genes H3, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, H22, H23, H25 and H26 to protect wheat from Hessian fly attack was assessed in natural field and under controlled laboratory conditions at INRAT-Kef Station. Results showed that Hessian fly was detected in 60.33% and 51.5% of all sampled durum and bread wheat fields, respectively. This pest was more frequent with a higher percentage of infestation in semi-arid regions. Indeed, during 2013, infestation rate attained 12.39% in Kef region against 0.9% registered in Bizerte region. In order to update information about the annual number of generations, we surveyed the population dynamic of Hessian fly in Kef region. Three generations of the fly were counted annually on wheat, with two complete and one incomplete generation. This insect affects host plant growth at different developmental stages. Plant height was the most affected parameter followed by shoot dry weight and tiller number. Field investigations on host resistance revealed that among the 16 tested resistance genes, and only three were strictly effective (H22, H25 and H26). The resistance genes H5, H9, H13 and H9H13 have also conferred high levels of protection against Hessian fly. This work indicated that H22, H25 and H26 genes could be incorporated into Tunisian wheat varieties and released to farmers to manage the threat due to Hessian fly attacks.     

Automated detection and classification of high frequency oscillations (HFOs) in human intracereberal EEG

Discrete high-frequency oscillations (HFOs) in the range of 80–500 Hz have previously been recorded from human epileptic brains using intracereberal EEG and seem to be a reliable biomarker of seizure onset zone in patients with intractable epilepsy. Visual marking of HFOs bursts is tedious, highly time-consuming particularly for analyzing long-term multichannel EEG recordings, inevitably subjective and can be error prone. Thus, the development of automatic, fast and robust detectors is necessary and crucial for HFOs investigation and for propelling their eventual clinical applications. This paper presents a proposed algorithm for detection and classification of HFOs, which is a combination of both smoothed Hilbert Huang Transform (HHT) and root mean square (RMS) feature. Performance evaluation in terms of sensitivity and false discovery rate (FDR) were respectively 90.72% and 8.23% related to process validation. Indeed, the proposed method was efficient in terms of high sensitivity in which the majority of HFOs visually identified by experienced reviewers was correctly detected, and had a lower FDR. This would mean that only a low rate of detected events was misclassified as candidate HFOs events. The presented software is extremely fast, suitable and can be considered a valuable clinical tool for HFOs investigation.