Saaïd Amzazi

Antioxidant activity of some Moroccan marine microalgae: Pufa profiles, carotenoids and phenolic content

In order to promote Moroccan natural resources, this study aims to evaluate the potential of microalgae isolated from Moroccan coastlines, as new source of natural antioxidants. Different extracts (ethanolic, ethanol/water and aqueous) obtained from 9 microalgae strains were screened for their in vitro antioxidant activity using DPPH free radical-scavenging assay. The highest antioxidant potentials were obtained in Dunalliela sp., Tetraselmis sp. and Nannochloropsis gaditana extracts. The obtained results indicate that ethanol extract of all microalgae strains exhibit higher antioxidant activity, when compared to water and ethanol/water extracts. Therefore, total phenolic and carotenoid content measurement were performed in active ethanol extracts. The PUFA profiles of ethanol extracts were also determined by GC/MS analysis. The studied microalgae strains displayed high PUFA content ranging from 12.9 to 76.9 %, total carotenoids content varied from 1.9 and 10.8mg/g of extract and total polyphenol content varied from 8.1 to 32.0mg Gallic acid Equivalent/g of extract weight. The correlation between the antioxidant capacities and the phenolic content and the carotenoids content were found to be insignificant, indicating that these compounds might not be major contributor to the antioxidant activity of these microalgae. The microalgae extracts exerting the high antioxidant activity are potential new source of natural antioxidants.

Gut microbiome compositional and functional differences between tumor and non-tumor adjacent tissues from cohorts from the US and Spain

Colorectal cancer (CRC) is the third most common cancer in the world and the second leading cause of cancer deaths in the US and Spain. The molecular mechanisms involved in the etiology of CRC are not yet elucidated due in part to the complexity of the human gut microbiota. In this study, we compared the microbiome composition of 90 tumor and matching adjacent tissue (adjacent) from cohorts from the US and Spain by 16S rRNA amplicon sequencing in order to determine the impact of the geographic origin on the CRC microbiome. Data showed a significantly (P < 0.05) higher Phylogenetic Diversity (PD) for the US (PD Adjacent = 26.3 ± 5.3, PD Tumor = 23.3 ± 6.2) compared to the Spanish cohort (PD Adjacent = 18.9 ± 5.9, PD Tumor = 18.7 ± 6.6) while no significant differences in bacterial diversity were observed between tumor and adjacent tissues for individuals from the same country. Adjacent tissues from the Spanish cohort were enriched in Firmicutes (SP = 43.9% and US = 22.2%, P = 0.0001) and Actinobacteria (SP = 1.6% and US = 0.5%, P = 0.0018) compared to US adjacent tissues, while adjacent tissues from the US had significantly higher abundances of Fusobacteria (US = 8.1% and SP = 1.5%, P = 0.0023) and Sinergistetes (US = 0.3% and SP = 0.1%, P = 0.0097). Comparisons between tumor and adjacent tissues in each cohort identified the genus Eikenella significantly over represented in US tumors (T = 0.024% and A = 0%, P = 0.03), and the genera Fusobacterium (T = 10.4% and A = 1.5%, P = <0.0001), Bulleida (T = 0.36% and A = 0.09%, P = 0.02), Gemella (T = 1.46% and A = 0.19%, P = 0.03), Parvimonas (T = 3.14% and A = 0.86%, P = 0.03), Campylobacter (T = 0.15% and A = 0.008%, P = 0.047), and Streptococcus (T = 2.84% and A = 2.19%, P = 0.05) significantly over represented in Spanish tumors. Predicted metagenome functional content from 16S rRNA surveys showed that bacterial motility proteins and proteins involved in flagellar assembly were over represented in adjacent tissues of both cohorts, while pathways involved in fatty acid biosynthesis, the MAPK signaling pathway, and bacterial toxins were over represented in tumors. Our study suggests that microbiome compositional and functional dissimilarities by geographic location should be taken in consideration when approaching CRC therapeutic options.

Scalable and cost-effective NGS genotyping in the cloud

BackgroundWhile next-generation sequencing (NGS) costs have plummeted in recent years, cost and complexity of computation remain substantial barriers to the use of NGS in routine clinical care. The clinical potential of NGS will not be realized until robust and routine whole genome sequencing data can be accurately rendered to medically actionable reports within a time window of hours and at scales of economy in the 10’s of dollars.ResultsWe take a step towards addressing this challenge, by using COSMOS, a cloud-enabled workflow management system, to develop GenomeKey, an NGS whole genome analysis workflow. COSMOS implements complex workflows making optimal use of high-performance compute clusters. Here we show that the Amazon Web Service (AWS) implementation of GenomeKey via COSMOS provides a fast, scalable, and cost-effective analysis of both public benchmarking and large-scale heterogeneous clinical NGS datasets.ConclusionsOur systematic benchmarking reveals important new insights and considerations to produce clinical turn-around of whole genome analysis optimization and workflow management including strategic batching of individual genomes and efficient cluster resource configuration.

In vitro Cytotoxic Effects and Antibacterial Activity of Moroccan Medicinal Plants Aristolochia longa and Lavandula multifida

Aims: The aim of this study was the evaluation of in vitro cytotoxic and antibacterial activities of organic extracts from Aristolochia longa and Lavandula multifida. Study Design: Evaluation of in vitro cytotoxic and antibacterial activities of extracts. Place and Duration of Study: Department of Biology (Faculty of Sciences), between July 2007 and July 2008. Methodology: The aerial parts were extracted by organic solvents (hexane, dichloromethane and methanol). The antibacterial activity of extracts was tested against Rhodococcus strains using the agar well diffusion method and cytotoxic activity was evaluated against three cancerous cell lines using the MTT assay. The chemical composition of extracts was determined using GC-MS. Results: The results show that the hexanic extract of A. longa (AH) and the dichloromethanic extract of A. longa (AD) present a good inhibitory effect on the three cancerous cell growth with 15 μg/ml≤IC50≤ 250 μg/ml and a total inhibitory effect on the bacterial growth with inhibition zone 30 mm at 50 mg/ml. Whereas, the extracts of L. multifida present less important inhibiting effects on the cell growth, in particular hexanic extract of L. multifida (LH) and dichloromethanic extract of Original Research Article Aneb et al.; EJMP, 16(2): 1-13, 2016; Article no.EJMP.28534 2 L. multifida (LD) with 115 μg/ml≤IC50≤ 300 μg/ml. These extracts are also active against the three strains of Rhodococcus, with more than 20 mm rings of inhibition at 50 mg/ml. The study of the chemical composition of each these species was undertaken by means of GC-MS. The phytochemical analysis of the extracts studied showed the presence of many chemical compounds which can explain these biological activities. These preliminary results suggest the presence in the extracts of compounds such as linoleic acid chloride; oleic acid and limonene-6-ol, pivalate for A. longa, and methyl linolenate; octadecane; oleic acid; 2,3,5,8-tetramethyldecane, phenol, 2methyl-5-(1-methylethyl) for L. multifida. Conclusion: This study deserves to be pursued to characterize better the asset compounds and to clarify their mechanism of action.

Screening of North African medicinal plant extracts for cytotoxic activity against tumor cell lines.

Aims: The aim of this study was to evaluate the in vitro cytotoxic activity and cellular effects of organic extracts and fractions of four plants; Inula viscosa, Ormenis eiriolepis (Asteraceae), Retama monosperma (Fabaceae) and Marrubium vulgare (Lamiaceae), all of them used in Moroccan traditional medicine. Methodology: The four plants were extracted using organic solvents and screened on a panel of human cancer cell lines including cell types from both solid and haematological cancer origin as well as non-transformed murine fibroblasts. Cell viability assays were Research Article European Journal of Medicinal Plants, 3(3): 310-332, 2013 311 performed with sixteen plant extracts. Sensitive cell lines were then exposed to increasing concentrations of the most efficient extracts in order to calculate IC50 values. Microscopy, flow cytometry and caspase activity assays were then performed in LN229, SW620 and PC-3 cell lines upon treatment to investigate the cell morphology, cell cycle distribution and cell death. Results: cell viability assays reveals that at least one extract from each plant was able to exert cytotoxic activity against the majority of cell lines tested, the IC50 values of the active extracts were in most cases ≤ 30 μg/ml. the study of the cellular effects of the most active extracts on LN229, SW620 and PC-3 cell lines shows their ability to promote cell cycle arrest and cell death. The data obtained herein support strongly the use of these plants by traditional healers for the treatment of cancer patients and could have some scientific support indicating the presence of bioactive compounds. Conclusion: The reported biological activity of these four medicinal plants used in traditional Moroccan medicine provides a starting point for forthcoming studies to determine the molecular basis of their activity and to identify the chemical compounds within the most active extracts responsible for their antitumoral effects.

Retama monosperma n-hexane extract induces cell cycle arrest and extrinsic pathway-dependent apoptosis in Jurkat cells

BackgroundRetama monosperma L. (Boiss.) or Genista monosperma L. (Lam.), locally named as “R’tam”, is an annual and spontaneous plant belonging to the Fabaceae family. In Morocco, Retama genus is located in desert regions and across the Middle Atlas and it has been widely used in traditional medicine in many countries. In this study, we show that Retama monosperma hexane extract presents significant anti-leukemic effects against human Jurkat cells.MethodsHuman Jurkat cells, together with other cell lines were screened with different concentrations of Retama monosperma hexane extract at different time intervals. Growth inhibition was determined using luminescent-based viability assays. Cell cycle arrest and apoptosis were measured by flow cytometry analysis. Combined caspase 3 and 7 activities were measured using luminometric caspase assays and immunoblots were performed to analyze expression of relevant pro- and anti-apoptotic proteins. GC-MS were used to determine the chemical constituents of the active extract.ResultsRetama monosperma hexane extract (Rm-HE) showed significant cytotoxicity against Jurkat cells, whereas it proved to be essentially ineffective against both normal mouse fibroblasts (NIH3T3) and normal lymphocytes (TK-6). Cytometric analysis indicated that Rm-HE promoted cell cycle arrest and apoptosis induction accompanied by DNA damage induction indicated by an increase in p-H2A.X levels. Rm-HE induced apoptosis was partially JNK-dependent and characterized by an increase in Fas-L levels together with activation of caspases 8, 3, 7 and 9, whereas neither the pro-apoptotic nor anti-apoptotic mitochondrial membrane proteins analyzed were significantly altered. Chemical identification analysis indicated that α-linolenic acid, campesterol, stigmasterol and sitosterol were the major bioactive components within the extract.ConclusionsOur data suggest that bioactive compounds present in Rm-HE show significant anti leukemic activity inducing cell cycle arrest and cell death that operates, at least partially, through the extrinsic apoptosis pathway.

Human papillomavirus detection in Moroccan patients with bladder cancer

INTRODUCTION Human papillomavirus (HPV) is associated with more human cancers than any other virus. Many studies have investigated the association between bladder cancer and HPV but the results remain controversial. The aim of the present study is to evaluate whether HPV have an etiological role in bladder carcinogenesis among Moroccan patients. METHODOLOGY Forty-eight fresh biopsies (43 bladder tumors and 5 non-tumor samples) were collected for this purpose. Nested PCR with the consensus MY09/MY11 and GP5+/GP6+ primers was performed to detect the presence of HPV L1 gene DNA. RESULTS The results showed that 52.4% of bladder cancer patients were positive for HPV. Subsequent DNA sequencing of positive cases of HPV revealed the presence of HPV16 in 95.5% of bladder tumor samples. The occurrence of HPV infection varies according to clinicopathological features, but there is no significant correlation between the viral infection and tumor stage or grade. In addition, statistical analysis demonstrated that there is no association between age or sex and HPV infection. CONCLUSION Our data indicate for the first time that bladder tumors from Moroccan patients harbor HR-HPV genotypes, especially HPV16, and thereby suggest that this virus may play a causative role in bladder cancer.

Pleistocene North African genomes link Near Eastern and sub-Saharan African human populations

Relationships among North Africans The general view is that Eurasians mostly descend from a single group of humans that dispersed outside of sub-Saharan Africa around 50,000 to 100,000 years ago. Present-day North Africans share a majority of their ancestry with present-day Near Easterners, but not with sub-Saharan Africans. To investigate this conundrum, Van de Loosdrecht et al. sequenced high-quality DNA obtained from bone samples of seven individuals from Taforalt in eastern Morocco dating from the Later Stone Age, about 15,000 years ago. The Taforalt individuals were found to be most closely related to populations from the Near East (Natufians), with a third of their ancestry from sub-Saharan Africa. No evidence was found for introgression with western Europeans, despite attribution to the Iberomaurusian culture. None of the present-day or ancient Holocene African groups are a good proxy for the sub-Saharan genetic component. Science, this issue p. 548 Ancient human genomes suggest dynamic interactions among Pleistocene African populations. North Africa is a key region for understanding human history, but the genetic history of its people is largely unknown. We present genomic data from seven 15,000-year-old modern humans, attributed to the Iberomaurusian culture, from Morocco. We find a genetic affinity with early Holocene Near Easterners, best represented by Levantine Natufians, suggesting a pre-agricultural connection between Africa and the Near East. We do not find evidence for gene flow from Paleolithic Europeans to Late Pleistocene North Africans. The Taforalt individuals derive one-third of their ancestry from sub-Saharan Africans, best approximated by a mixture of genetic components preserved in present-day West and East Africans. Thus, we provide direct evidence for genetic interactions between modern humans across Africa and Eurasia in the Pleistocene.

COSMOS: cloud enabled NGS analysis

Background The dramatic fall of next generation sequencing (NGS) cost in recent years positions the price in range of typical medical testing, and thus whole genome analysis (WGA) may be a viable clinical diagnostic tool. Modern sequencing platforms routinely generate petabyte data. The current challenge lies in calling and analyzing this large-scale data, which has become the new time and cost rate-limiting step.

Rifoligotyping assay: an alternative method for rapid detection of rifampicin resistance in Mycobacterium tuberculosis isolates from Morocco

One of the greatest threats to global tuberculosis (TB) control is the growing prevalence of drug resistant strains. In the past decades, considerable efforts have been made upon the development of new molecular technologies and methodologies for detection of drug resistance in Mycobacterium tuberculosis (MTB). A sensitive, specific reverse line blot assay, called rifoligotyping (RIFO), for the detection of genotypic resistance to rifampicin (RIF), was designed and evaluated. RIFO includes oligonucleotide probes specific for wild-type and mutant sequences, allowing specific and sensitive detection of both genotypes in a single assay. The RIFO was applied on 500 MTB isolates from Morocco. The results of the RIFO showed a good sensitivity (90.9%) and high specificity (100%); the positive and negative predictive values were 100% and 96.1%, respectively. This rapid, simple, economical assay provides a practical alternative for RIF genotyping, especially in low-income countries, to improve TB control and management.