Anticancer Effects of New Ceramides Isolated from the Red Sea Red Algae Hypnea musciformis in a Model of Ehrlich Ascites Carcinoma: LC-HRMS Analysis Profile and Molecular Modeling

Different classes of phytochemicals were previously isolated from the Red Sea algae Hypnea musciformis as sterols, ketosteroids, fatty acids, and terpenoids. Herein, we report the isolation of three fatty acids—docosanoic acid 4, hexadecenoic acid 5, and alpha hydroxy octadecanoic acid 6—as well as three ceramides—A (1), B (2), and C (3)—with 9-methyl-sphinga-4,8-dienes and phytosphingosine bases. Additionally, different phytochemicals were determined using the liquid chromatography coupled with electrospray ionization high-resolution mass spectrometry (LC-ESI-HRMS) technique. Ceramides A (1) and B (2) exhibited promising in vitro cytotoxic activity against the human breast adenocarcinoma (MCF-7) cell line when compared with doxorubicin as a positive control. Further in vivo study and biochemical estimation in a mouse model of Ehrlich ascites carcinoma (EAC) revealed that both ceramides A (1) and B (2) at doses of 1 and 2 mg/kg, respectively, significantly decreased the tumor size in mice inoculated with EAC cells. The higher dose (2 mg/kg) of ceramide B (2) particularly expressed the most pronounced decrease in serum levels of vascular endothelial growth factor -B (VEGF-B) and tumor necrosis factor-α (TNF-α) markers, as well as the expression levels of the growth factor midkine in tumor tissue relative to the EAC control group. The highest expression of apoptotic factors, p53, Bax, and caspase 3 was observed in the same group that received 2 mg/kg of ceramide B (2). Molecular docking simulations suggested that ceramides A (1) and B (2) could bind in the deep grove between the H2 helix and the Ser240-P250 loop of p53, preventing its interaction with MDM2 and leading to its accumulation. In conclusion, this study reports the cytotoxic, apoptotic, and antiangiogenic effects of ceramides isolated from the Red Sea algae Hypnea musciformis in an experimental model of EAC.     

Sequence analysis of the internal transcribed spacers (ITSs) region of the nuclear ribosomal DNA (nrDNA) in fig cultivars (Ficus carica L.)

Sequences of the internal transcribed spacers (ITSs) of nuclear ribosomal DNA (nrDNA) were analysed in a set of Tunisian fig (Ficus carica L.) cultivars. The size of the spacers sequences ranged from 200 to 279 bases for ITS1 and from 253 to 314 bases for ITS2. Variation of GC contents has been also observed and scored as 59–68% and 55–68% for ITS1 and ITS2, respectively. This data exhibited the presence of polymorphism among cultivars. The intra-specific variability level of the ITS sequences proved a variation both in the length and in the sequences studied. In fact, ITS1 and ITS2 sequences were considered as a useful tool to establish genetic relationships among cultivars. Our results indicate that the diversity detected among closely related genotypes supported strongly the efficiency of ITS sequences for establishing relationships between cultivars. ITS2 seems to be relatively more informative than ITS1 regarding length or GC contents. Considerable genetic diversity was observed among fig at intra and inter-cultivars levels. Two polyclonal varieties were identified. In addition, data proved that a typically continuous genetic diversity characterizes the local fig germplasm. The topology of the derived dendrogram strongly supported this assumption. In fact, genotypes are clustered independently from their geographical origin or the sex of trees suggesting a narrow genetic basis among the ecotypes studied in spite of their phenotypic distinctiveness. Implications of these results for management of fig germplasm collections are discussed.     

EST-SSR based estimates on functional genetic variation in a barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) collection from Egypt

Genetic diversity was investigated in a set of 27 barley genotypes, originated from Egypt, using 23 expressed sequence tag (EST)-derived simple sequence repeats (eSSRs), representatives of the seven barley chromosomes. Ninety-five alleles were detected among all the barley accessions. For 22 polymorphic eSSRs, the number of alleles per locus varied from 2 to 12, with a mean of 4.318 alleles per locus. The lowest and the largest number of alleles per locus among the seven homeologous groups was observed in homeologous group 5H and 4H with 3.00 and 6.33, respectively. The gene diversity increased as the number of alleles increased. Gene diversity for 22 ESTs loci varied from 0.137 for GBM1404 to 0.896 for GBM1015 with an average of 0.563. A significant correlation coefficient between gene diversity and the number of alleles was high, r = 0.741 (P < 0.01). Cluster analysis was conducted based on eSSRs data to group the barley genotypes and to construct a dendrogram. Four groups can be distinguished by truncating the dendrogram at gs value of 0.77.     

Chemical composition of the volatile extract and antioxidant activities of the volatile and nonvolatile extracts of Egyptian corn silk (Zea mays L.)

A total of 36 compounds, which comprised 99.4% of the extract, were identified by gas chromatography and mass spectrometry (GC-MS) in the volatile dichloromethane extract obtained from Egyptian corn silk. The main constituents of the volatile extract were cis-alpha-terpineol (24.22%), 6,11-oxidoacor-4-ene (18.06%), citronellol (16.18%), trans-pinocamphone (5.86%), eugenol (4.37%), neo-iso-3-thujanol (2.59%), and cis-sabinene hydrate (2.28%). Dried Egyptian corn silk was also directly extracted with petroleum ether, ethanol, and water. All extracts from solvent extraction and the volatile extract described above exhibited clear antioxidant activities at levels of 50-400 microg/mL in the 2,2-diphenyl-1-picrylhydrazyl (DPPH)/linoleic acid assay. The ethanol extract inhibited DPPH activity by 84% at a level of 400 microg/mL. All samples tested via the beta-carotene bleaching assay also exhibited satisfactory antioxidant activity with clear dose responses. This study indicates that corn silk could be used to produce novel natural antioxidants as well as a flavoring agent in various food products.     

Effect of Khat (Catha edulis Forsk) extract on testicular maturation in pre-pubertal and pubertal rats: A morphological and biochemical study

The present study aimed at analysing the effect of Khat plant extract on rat testicular development. Thirty-two weaned male albino rats (4 weeks old) were divided into four groups consisting of eight animals each. While control animals received normal saline, rats of groups I, II and III received 100, 200 and 300 mg Khat extract per kg body weight dissolved in distilled water by oral gavage daily for 8 weeks, respectively. Blood samples were collected in separate heparinized tubes by cardiac puncture from each rat and processed for measuring plasma levels of reproductive hormones LH, FSH, testosterone and prolactin. Five-µm sections were stained with haematoxylin and eosin and examined by light microscope. Some sections were immunostained for protamine-1 representing a biomarker for intact sperm differentiation. The present study clearly demonstrated that Khat extract has a pronounced effect on testicular maturation of developing albino rats at both the morphological and functional levels. Khat-treated groups revealed a significantly low serum testosterone level and severe impairment of spermatogenesis when compared with control animals. The current findings also verified, for the first time, that the final stages of sperm maturation (spermiogenesis) were strongly impaired after administration of Khat extract to experimental rats particularly at a higher dose (300 mg/kg body weight). This was proved by the very weak, if any, expression of protamine-1 in the maturing spermatids in Khat-treated rats.     

Amelioration of sperm fertilizability,thyroid activity,oxidative stress,and inflammatory cytokines in rabbit bucks treated with phytogenic extracts

This study investigated the beneficial effect of phytogenic extracts on semen quality, reproductive hormones, thyroid activity, immunity, hepatic antioxidant activity, and fertility in rabbit bucks. We divided 70 bucks into seven groups (10 in each). Group 1 was fed a basal diet (control); groups 2, 3, and 4 were fed the control diet with 30, 60, and 90 mg/kg of turmeric, respectively; and groups 5, 6, and 7 were fed the control diet with 50, 75, and 100 mg/kg of garlic extract, respectively, for 8 weeks. Rectal and skin temperatures decreased, while follicle-stimulating hormone, luteinizing hormone, triiodothyronine, thyroxine, testosterone, immunoglobulin M, tumor necrosis factor-alpha, and interleukin-6 in blood serum and glutathione peroxidase in the liver increased in all groups (p < .05). Garlic extract (100 mg/kg diet) increased adenosine triphosphate and glutathione in the liver tissues. All treatments significantly increased net semen volume, percentages of progressive motility, livability, curled tail, and intact acrosomes of spermatozoa, sperm cell concentration, and outputs of total and motile spermatozoa, while significantly decreased percentage of sperm abnormality. In conclusion, dietary supplementation of turmeric or garlic extract can be used as a suitable tool for enhancing the hepatic antioxidant activity, immunity, and semen quality in rabbit bucks.     

Mapping QTLs in breeding for drought tolerance in maize (Zea mays L.)

Summary Grain yield in the maize (Zea mays L) plant is sensitive to drought in the period three weeks either side of flowering. Maize is well-adapted to the use of restriction fragment length polymorphisms (RFLPs) to identify a tight linkage between gene(s) controlling the quantitative trait and a molecular marker. We have determined the chromosomal locations of quantitative trait loci (QTLs) affecting grain yield under drought, anthesis-silking interval, and number of ears per plant. The F₃ families derived from the cross SD34(tolerant) × SD35 (intolerant) were evaluated for these traits in a two replicated experiment. RFLP analysis of the maize genome included non-radioactive DNA-DNA hybridization detection using chemiluminescence. To identify QTLs underlying tolerance to drought, the mean phenotypic performances of F₃ families were compared based on genotypic classification at each of 70 RFLP marker loci. The genetic linkage map assembled from these markers was in good agreement with previously published maps. The phenotypic correlations between yield and other traits were highly significant. In the combined analyses, genomic regions significantly affecting tolerance to drought were found on chromosomes 1,3,5,6, and 8. For yield, a total of 50% of the phenotypic variance could be explained by five putative QTLs. Different types of gene action were found for the putative QTLs for the three traits.