Assessment of impacts of industrial effluents on physico-chemical and microbiological qualities of irrigation water of the Fez Rriver, Morocco.

Global water consumption has grown twice as fast as the population. Wastewater is therefore a valuable and renewable source and provides additional water for priority uses. Wastewater can also be a source of pollution; thus, its physico-chemical and biological compositions can present major risks to the environment and human health. The objective of this study was to assess the status of irrigation waters in terms of salinization, accumulation of metallic elements, and microbiological contamination by parasites and pathogenic bacteria. The study focused on the surface water of Oued Fès used for irrigation located downstream of the industrial zone of Doukkarat and upstream of the industrial zone of Ain Noukbi (wastewater) before the confluence with the Oued Sebou, as well as on the treated wastewater of the wastewater treatment plant. The physico-chemical and microbiological analyses were carried out in two periods: summer and winter. Metals were analyzed by ICP-AES. The chemical and bacteriological quality of the wastewater and treated wastewater was found to be poor. These were characterized by organic pollution, including biodegradable pollutants, while upstream the organic residues were not biodegradables. COD, BOD5, Kjeldahl nitrogen, as well as chloride ion (Cl-) are above the standard values. The highest concentrations of Cd, at 850 µg/l, Cu, at 690 µg/l and Mn, at 470 µg/l, largely exceed the international standards and requirements. In addition to fecal contamination, characterized by total coliforms and thermo-tolerant coliforms, other pathogens were present, including helminth eggs, both in the wastewater and in the treated wastewater. Other pathogens, such as Vibrio cholera, were found at all three sites whether in winter or summer, with the exception of the downstream of Oued Fez in winter. As for Salmonella, it was present in treated wastewater during the winter only. The water used for irrigation upstream of Oued Fez and the treated wastewater have poor to very poor quality. Therefore, for a better use of these waters, it is necessary to ensure their regular treatment in order to minimize the impacts on the environment and human health.

Tomentosin Induces Telomere Shortening and Caspase-Dependant Apoptosis in Cervical Cancer Cells.

Tomentosin, a natural sesquiterpene lactone purified from of Inula viscosa L., was investigated for its anti-proliferative, telomere shortening, and apoptotic effects on human cervical cancer HeLa and SiHa cell lines. Tomentosin was found to inhibit the growth of SiHa and HeLa cell lines in dose and time-dependent manner (IC50 values of 7.10 ± 0.78 µM and 5.87 ± 0.36 µM, respectively after 96 h of treatment). As evidenced by TTAGGG telomere length assay, tomentosin target specifically the telomeric overhang lengthening. This was confirmed by the evaluation of the cytotoxic effects of tomentosin in the foetal fibroblast Wi38 and JW10 cells which were derived from Wi38 and express hTERT, the telomerase catalytic subunit. We found that JW10 cells are 4.7-fold more sensitive to tomentosin which argues for telomere as its specific target. Furthermore, we found that tomentosin mediate this cytotoxic effect by inducing apoptosis and cell cycle arrest at G2/M phase. Morphological features of treated cells, as evidenced by Hoechst 33324 staining, revealed that the cytotoxic effect was due to induction of apoptosis. This was accompanied by pro-caspase-3 cleavage, an increase in caspase-3 activity and a cleavage of poly (ADP-ribose) polymerase (PARP). Moreover, tomentosin induced a decrease in mitochondrial membrane potential (ΔΨm) and an increase in reactive oxygen species (ROS), accompanied by a decrease in Bcl-2 expression. This indicates that tomentosin-induced apoptosis may involve a mitochondria-mediated signaling pathway. This study provides the first evidence that tomentosin targets telomere machinery and induces apoptosis in cervical cancer cells. The molecular mechanism underlying tomentosin-induced apoptosis may involve a mitochondria-mediated signaling pathway. J. Cell. Biochem. 118: 1689-1698, 2017. © 2016 Wiley Periodicals, Inc.

Inula Viscosa Extracts Induces Telomere Shortening and Apoptosis in Cancer Cells and Overcome Drug Resistance.

Telomerase is activated in human papillomavirus (HPV) positive cervical cancer and targeting telomeres offers a novel anticancer therapeutic strategy. In this study, the telomere targeting properties, the cytotoxic as well as the pro-apoptotic effects of hexane (IV-HE) and dichloromethane (IV-DF) fractions from Inula viscosa L. extracts were investigated on human cervical HeLa and SiHa cancer cells. Our data demonstrate that IV-HE and IV-DF extracts were able to inhibit cell growth in HeLa and SiHa cells in a dose-dependent manner and studied resistant cell lines exhibited a resistance factor less than 2 when treated with the extracts. IV-HE and IV-DF extracts were able to inhibit telomerase activity and to induce telomere shortening as shown by telomeric repeat amplification protocol and TTAGGG telomere length assay, respectively. The sensitivity of fibroblasts to the extracts was increased when telomerase was expressed. Finally, IV-HE and IV-DF were able to induce apoptosis as evidenced by an increase in annexin-V labeling and caspase-3 activity. This study provides the first evidence that the IV-HE and IV-DF extracts from Inula viscosa L. target telomeres induce apoptosis and overcome drug resistance in tumor cells. Future studies will focus on the identification of the molecules involved in the anticancer activity.

Assessment of anti-hyperglycemic and anti-hyperlipidemic effects of thiazolidine-2,4-dione derivatives in HFD-STZ diabetic animal model.

Type 2 diabetes mellitus (T2DM) is a chronic endocrine/metabolic disorder characterized by elevated postprandial and fasting glycemic levels that result in disturbances in primary metabolism. In this study, we evaluated the metabolic effects of thiazolidine-2,4-dione derivatives in Wistar rats and Swiss mice that were fed a high-fat diet (HFD) for 4 weeks and received 90 mg/kg of streptozotocin (STZ) intraperitoneally as a T2DM model. The HFD consisted of 17% carbohydrate, 58% fat, and 25% protein, as a percentage of total kcal. The thiazolidine-2,4-dione derivatives treatments reduced fasting blood glucose (FBG) levels by an average of 23.98%-50.84%, which were also improved during the oral starch tolerance test (OSTT). Treatment with thiazolidine-2,4-dione derivatives also improved triglyceride (TG), low-density lipoprotein cholesterol (LDL-c), and total cholesterol levels (P < 0.05). The treatment intake has also shown a significant effect to modulate the altered hepatic and renal biomarkers. Further treatment with thiazolidine-2,4-dione derivatives for 28 days significantly ameliorated changes in appearance and metabolic risk factors, including favorable changes in histopathology of the liver, kidney, and pancreas compared with the HFD/STZ-treated group, suggesting its potential role in the management of diabetes. Thiazolidine-2,4-dione derivatives are a class of drugs that act as insulin sensitizers by activating peroxisome proliferator-activated receptor-gamma (PPAR-γ), a nuclear receptor that regulates glucose and lipid metabolism. The results of this study suggest that thiazolidine-2,4-dione derivatives may be a promising treatment option for T2DM by improving glycemic control, lipid metabolism, and renal and hepatic function.

Comprehensive behavioural assessment of TERT in bladder cancer.

BACKGROUND: Telomerase activity plays a crucial role in cancer development and progression. Thus, telomerase activation through the interplay of mutations and epigenetic alterations in the telomerase reverse transcriptase (TERT) promoter may provide further insight into bladder cancer induction and progression. METHODS: In this study 100 bladder tumour tissues were selected, and four molecular signatures were analysed: THOR methylation status, TERT promotor mutation, telomere length, and TERT expression. RESULTS: In our study, 88% of bladder cancer patients had an hypermethylation of the THOR region and 60% had mutations in the TERT promoter region. TERT promoter methylation was observed in all stages and grades of bladder cancer. While, TERT promoter mutations were detected in advanced stages and grades. In our cohort, high levels of TERT expression and long telomeres have been found in noninvasive cases of bladder cancer, with a significant association between TERT expression and Telomere length. Interestingly, patients with low TERT expression and cases with long telomeres had significantly longer Disease-free survival and overall survival. CONCLUSION: The methylation and mutations occurring in the TERT promoter are implicated in bladder carcinogenesis, offering added prognostic and supplying novel insight into telomere biology in cancer.

Analysis of mutations in the E6 oncogene of human papillomavirus 16 in cervical cancer isolates from Moroccan women.

BACKGROUND: Worldwide, cervical cancer is the second most common cancer in women. High-risk human papillomavirus (HPV) play a crucial role in the etiology of cervical cancer and the most prevalent genotype is HPV16. HPV 16 intratypic variants have been reported to differ in their prevalence, biological and biochemical properties. The present study was designed to analyze and identify HPV type 16 E6 variants among patients with cervical cancer in Morocco. METHODS: A total of 103 HPV16 positive samples were isolated from 129 cervical cancer cases, and variant status was subsequently determined by DNA sequencing of the E6 gene. RESULTS: Isolates from patients were grouped into the European (E), African (Af) and North-American (NA1) phylogenetic clusters with a high prevalence of E lineage (58.3%). The Af and NA1 variants were detected in 31.1% and 11.6% of the HPV16 positive specimens, respectively, whereas, only 3% of cases were prototype E350T. No European-Asian (EA), Asian (As) or Asian-American (AA) variants were observed in our HPV16-positive specimens. At the amino acid level, the most prevalent non-synonymous variants were L83V (T350G), H78Y (C335T), E113D (A442C), Q14D (C143G/G145T) and R10I (G132T), and were observed respectively in 65%, 41.8%, 38.8%, 30.1% and 23.3% of total samples.Moreover, HPV16 European variants were mostly identified in younger women at early clinical diagnosis stages. Whereas, HPV16 Af variants were most likely associated with cervical cancer development in older women with pronounced aggressiveness. CONCLUSION: This study suggests a predominance of E lineage strains among Moroccan HPV 16 isolates and raises the possibility that HPV16 variants have a preferential role in progression to malignancy and could be associated with the more aggressive nature of cervical cancer.

Biological, toxicological and molecular docking evaluations of isoxazoline-thiazolidine-2,4-dione analogues as new class of anti-hyperglycemic agents.

In this work, three isoxazoline-thiazolidine-2,4-dione derivatives were synthesized and characterized by FT-IR, 1H-NMR, 13C-NMR and ESI-MS spectrometry. All compounds have been investigated for their α-amylase and α-glucosidase inhibitory activities. In vitro enzymatic evaluation revealed that all compounds were inhibitory potent against α-glucosidase with IC50 values varied from 40.67 ± 1.81 to 92.54 ± 0.43 µM, and α-amylase with IC50 in the range of 07.01 ± 0.02 to 75.10 ± 1.06 µM. One of the tested compounds were found to be more potent inhibitor compared to other compounds and standard drug Acarbose (IC50 glucosidase= 97.12 ± 0.35 µM and IC50 amylase= 2.97 ± 0.01 µM). All compounds were then evaluated for their acute toxicity in vivo and shown their safety at a high dose with LD > 2000mg/kg BW. A cell-based toxicity evaluation was performed to determine the safety of compounds on liver cells, using the MTT assay against HepG2 cells, and the results shown that all compounds have non-toxic impact against cell viability and proliferation compared to reference drug (Pioglitazone). Furthermore, the molecular homology analysis, SAR and the molecular binding properties of compound with the active site of α-amylase and α-glucosidase were confirmed through computational analysis. This study has identified the inhibitory potential of a new class of synthesized isoxazoline-thiazolidine-2,4-dione derivatives in controlling both hyperglycemia and type 2 diabetes mellitus without any hepatic toxicity.Communicated by Ramaswamy H. Sarma.

The Evaluation of Vascular Endothelial Growth Factor A (VEGFA) and VEGFR2 Receptor as Prognostic Biomarkers in Bladder Cancer.

Vascular endothelial growth factor (VEGF) and its receptors (VEGFR1 and VEGFR2) are the most important tissue factors involved in tumor growth and angiogenesis. The aim of this study was to evaluate the promoter mutational status of VEGFA and the expression levels of VEGFA, VEGFR1, and VEGFR2 in bladder cancer (BC) tissues and to correlate the results with the clinical-pathological parameters of BC patients. A total of 70 BC patients were recruited at the Urology Department of the Mohammed V Military Training Hospital in Rabat, Morocco. Sanger sequencing was performed to investigate the mutational status of VEGFA, and RT-QPCR was used to evaluate the expression levels of VEGFA, VEGFR1, and VEGFR2. Sequencing of the VEGFA gene promoter revealed the presence of -460T/C, -2578C/A, and -2549I/D polymorphisms, and statistical analyses showed a significant correlation between -460T/C SNP and smoking (p = 0.02). VEGFA and VEGFR2 expressions were significantly up-regulated in patients with NMIBC (p = 0.003) and MIBC (p = 0.03), respectively. Kaplan-Meier analyses showed that patients with high VEGFA expression had significantly longer disease-free survival (p = 0.014) and overall survival (p = 0.009). This study was very informative, showing the implication of VEGF alterations in BC, suggesting that VEGFA and VEGFR2 expressions could be promising biomarkers for the better management of BC.

Evaluation of DNA methylation in promoter regions of hTERT, TWIST1, VIM and NID2 genes in Moroccan bladder cancer patients.

Promoter hypermethylation have been reported to play a key role in bladder cancer development and progression. The aim of this study is to evaluate the methylation status of hTERT, TWIST1, VIM and NID2 genes in bladder cancer. The methylation status was evaluated using the Methylation-Specific PCR (MSP) approach on 70 tumour biopsies from Moroccan bladder cancer patients. Overall, methylation frequencies of hTERT, TWIST1, VIM and NID2 genes, were 90%, 85.71%, 67.14% and 67.14%, respectively. Hypermethylation of all studied genes was found in all pathological grades and stages of bladder cancer. Nevertheless, statistical analysis showed no significant association between promoter methylation of hTERT, TWIST1, VIM and NID2 genes and tumours stage/grade (p value >0.05). Moreover, we have investigated the association between the methylation pattern of selected genes and the treatment outcome in a sub-group of non-muscle-invasive bladder cancer cases (52/70). Hypermethylation of hTERT, TWIST1, VIM and NID2 was detected in 83.34%; 66.67%; 83.34% and 58.34% of recurrent cases, respectively, and in 80%; 80%; 80% and 60% of progressive cases, respectively. Statistical analysis highlighted a significant association between TWIST1 hypermethylation and tumour recurrence (p = 0.041<0.05). Our results indicate that hypermethylation of hTERT, TWIST1, VIM and NID2 genes is a frequent epigenetic event in bladder cancer and could be a promising therapeutic target to prevent bladder cancer progression and metastasis.

Apoptosis-mediated anti-proliferative activity of Calligonum comosum against human breast cancer cells, and molecular docking of its major polyphenolics to Caspase-3.

Due to poor diagnosis breast cancer in women has emerged as the most common cause of death disease in developing countries. Medicinal plants have been used for thousands of years and can be useful in healthcare, especially in developing countries. Ethanol extracts of leaves of fire bush or arta (Calligonum comosum; EECC), exhibited significant anticancer potencies against two breast cancer cell lines, MCF-7 and MDA 231. These in vitro effects of EECC indicated potential anticancer activities that were determined to be specific since minimal toxicity was recorded against MCF-12, a non-cancerous breast cell line used as a reference. EECC also induced cell cycle arrest in MCF-7 and MDA 231 as revealed by the increased proportions of sub-G1 cells. Fluorescence-activated cell sorter analysis (FACS), utilizing double staining by annexin V-FITC/propidium iodide, revealed that the observed cytotoxic effects were mediated via apoptosis and necrosis. FACS measurement of thegreater in fluorescence intensity, linked with oxidation of DCFH to DCF, revealed that apoptosis was attributable to production of free radicals. EECC-mediated apoptosis was further validated by observation of up-regulation in the "executioner" enzyme, caspase 3. The current findings reveal that EECC exhibits significant, selective cytotoxicity to breast cancer cells, that proceeds via the generation of ROS, which culminates in apoptosis. The anti-proliferative effects of EECC weres further verified by use of a structure-based, virtual screening between its major bioactive polyphenolic constituents and the apoptosis executioner marker enzyme, caspase-3. Based on their glide score values against the active site of caspase 3, some phyto-constituents present in EECC, such as DL-alpha-tocopherol and campesterol, exhibited distinctive, drug-like potential with no predicted toxicity to non-target cells. Taken together, the usefulness of natural phenolic and flavonoid compounds contained in Calligonum comosum were suggested to be potent anticancer agents.

Stipa tenacissima L.: A New Promising Source of Bioactive Compounds with Antioxidant and Anticancer Potentials.

BACKGROUND: Stipa tenacissima L. (S. tenacissima), called Esparto grass, is a cultivated species used for industrial purposes, including textile production. This species has never been studied for its medical potential before, nor has it been used in traditional medicines. It is thus fitting that the present study aimed to investigate the pharmacological potential of S. tenacissima. To achieve this goal, this work was conducted to study the chemical composition, antioxidant properties, and antiproliferative effects of S. tenacissima against cancerous cell lines, including the human colorectal adenocarcinoma cell line (HT-29) and human breast adenocarcinoma cell line (MDA-MB-231). Fractionation and characterization of S. tenacissima extract showed the presence of promising bioactive fractions. The fractions obtained from S. tenacissima extract exhibited interesting antioxidant properties, with IC50 values ranging from 1.26 to 1.85 mg/mL. All fractions, such as F1, F2, F3, and F4, induced an important antiproliferative effect on the cancer cell lines MDA-MB-231, scoring IC50 values ranging from 63.58 ± 3.14 to 99.880 ± 0.061 µg/mL. These fractions (F1, F2, F3, and F4) also exhibited a potent antiproliferative effect versus HT-29 cell lines, with IC50 values ranging from 71.50 ± 4.97 to 87.500 ± 1.799 µg/mL. Therefore, S. tenacissima could constitute a new natural source of bioactive compounds that can be used for therapeutic purposes to fight cancer and free radical damage.

Immune Checkpoint and Telomerase Crosstalk Is Mediated by miRNA-138 in Bladder Cancer.

BACKGROUND: Tumor recurrence and progression in non-muscle invasive bladder cancer (NMIBC), therapy failure, and severe side effects in muscle invasive bladder cancer (MIBC) are the major challenges in the clinical management of bladder cancer (BC). Here, we identify new molecular targetable signatures to improve BC patients' stratification and the outcome of current immunotherapies. MATERIAL AND METHODS: In a prospective cohort of 70 BC patients, we assessed the genetic and molecular regulation of TERT in maintaining telomere length in parallel to immune checkpoint and microRNA expression. RESULTS: TERT was undetectable in healthy bladder tissues but upregulated in invasive BC stages and high tumor grade. Its expression was linked with the combined effect of the C250T mutation and THOR hypermethylation, associated with progressing tumors and maintaining of telomere length. In the same cohort, PD-L1 scored highest in NMIBC, while PD-L2 was upregulated in MIBC. We also show that miR-100-5p and 138-5p were highly expressed in healthy bladder specimens and cell line, while expression decreased in the BC tissues and BC cell lines. In line with the binding prediction for these miRNAs on target genes, miRs 100-5p and 138-5p expression strongly inverse correlated with TERT, PD-L1, and PD-L2 expression, but not PD1. CONCLUSION: We identify a loop involving TERT, PD1-ligands, and miR-138-5p in BC, that might represent not only a useful biomarker for improved diagnosis and patients' stratification but also as a promising axis that might be therapeutically targeted in situ.

Phytochemical Study on Antioxidant and Antiproliferative Activities of Moroccan Caralluma europaea Extract and Its Bioactive Compound Classes.

BACKGROUND: Caralluma europaea (C. europaea) is a medicinal plant used in Moroccan popular medicine. Objective of the Study. The present work was aimed at identifying the chemical composition and the antioxidant and antiproliferative activities of hydroethanolic and bioactive compound classes of C. europaea) is a medicinal plant used in Moroccan popular medicine. Materials and Methods. The chemical composition was analyzed using HPLC. The antioxidant power was determined using both DPPH and FRAP assays. The antiproliferative activity was effectuated against cancerous cells using WST-1. RESULTS: The chemical analysis showed the presence of bioactive constituents such as quercetin, myricetin, and hesperetin. The polyphenol and flavonoid contents were estimated at 51.42 mg GA/g and 20.06 mg EQ/g, respectively. The EC50 values of FRAP assay of hydroethanolic, flavonoid, saponin, and mucilage extracts were 5.196 mg/ml, 4.537 mg/ml, 3.05 mg/ml, and 6.02 mg/ml, respectively. The obtained IC50 values with the DPPH test were 1.628 mg/ml, 1.05 mg/ml, 1.94 mg/ml, and 9.674 mg/ml, respectively. Regarding MDA-MB-231, saponins were highly effective even with the lowest concentration (15.62 µg/ml). The flavonoids decreased the cell viability with IC50 values of 43.62 ± 0.06 µg/ml). The flavonoids decreased the cell viability with IC50 values of 43.62 ± 0.06 µg/ml). The flavonoids decreased the cell viability with IC50 values of 43.62 ± 0.06 . CONCLUSION: The present results suggest that C. europaea) is a medicinal plant used in Moroccan popular medicine.

Phytochemical Analysis, Cytotoxic, Antioxidant, and Antibacterial Activities of Lichens.

BACKGROUND: Lichens present a complex symbiotic relationship between a filamentous fungus, photoautotrophic partner (algae or cyanobacteria), and bacterial community. The Objective of the Study. This study aimed at investigating the chemical composition and cytotoxic, antioxidant, and antimicrobial activities of acetone extracts of Moroccan Evernia prunastri (E. prunastri), Ramalina farinacea (R. farinacea), and Pseudevernia furfuracea (P. furfuracea). Materials and Methods. The phytochemical analysis was carried out by HPLC-UV. The cytotoxic effect was assessed on human prostate cancer (22RV1), human colon carcinoma (HT-29), human hepatocellular carcinoma (Hep-G2), and Hamster ovarian cancer (CHO) cells lines by WST1 assay. The antioxidant power was assessed by DPPH and FRAP assays. The antibacterial effect was obtained using the broth microdilution method. RESULTS: The findings of phytochemical analysis showed that the lichens studied possess interesting bioactive molecules such as physodalic acid, evernic acid, and usnic acid, as well as protocetraric acid. According to the American National Cancer Institute guidelines, the WST-1 test showed that all crude extracts did not show significant cytotoxic effects against all concerous cell lines, and IC50 values ranged from 42.30 to 140.24 µg/mL. Regarding the antioxidant activity, P. furfuracea extract showed the highest free-radical-scavenging ability (IC50 = 498.40 µg/mL). The most potent antibacterial extract was recorded for P. furfuracea extract with a minimum inhibitory concentration (MIC) ranging from 0.039 to 0.31 mg/mL. CONCLUSION: In this research work, we report that the studied lichen extracts exhibit an important biological effect, supporting that lichens represent a hopeful source of original natural products for the research of new bioactive molecules having a pharmaceutical interest.

Status of p16(INK4a) and E-cadherin gene promoter methylation in Moroccan patients with cervical carcinoma.

Aberrant methylation of tumor suppressor gene promoters has been extensively investigated in cervical cancer. Transcriptional silencing, as a main consequence of hypermethylation of CpG islands, is the predominant mechanism of p16(INK4a) and E-cadherin gene inactivation in malignant epithelial tumors. This study was conducted to evaluate the promoter methylation status of p16(INK4a) and E-cadherin genes in 22 specimens of cervical carcinomas, four cervical cancer cell lines (HeLa, SiHa, Caski, C33A), and 20 human papillomavirus negative specimens, obtained from normal cervical swabs, using the methylation-specific PCR approach. Hypermethylation of the 5' CpG island of the p16(INK4a) and E-cadherin genes were found in 13 (59.1%) and 10 (45.5%) of 22 cervical cancer samples, respectively. Furthermore, our findings did not show any correlation between promoter methylation of p16(INK4a) and E-cadherin genes and clinicopathological parameters, including HPV infection, phenotypic distribution, and stage of the disease. However, hypermethylation of E-cadherin gene promoter appears to be age related in cervical cancer, whereas the frequency of aberrant methylation of p16(INK4a) gene promoter is unchanged according to the age of patients. Thus, caution must be made to use these markers in the diagnosis of cervical cancer. However, dietary or pharmaceutical agents that can inhibit these epigenetic events may prevent or delay the development of cervical cancer.

Promising Physical, Physicochemical, and Biochemical Background Contained in Peels of Prickly Pear Fruit Growing under Hard Ecological Conditions in the Mediterranean Countries.

BACKGROUND: Prickly pear (Opuntia spp.), called Barbary fig, is a cultivated species springing from family Cactaceae. It is native to Mexico and has been naturalized in other continents, especially the Mediterranean countries (North Africa). The aim of the study was to investigate the physical, physicochemical, and biochemical criteria of peels of three Moroccan prickly pear varieties (Aakria, Derbana, and Mles) growing in the Rhamna regions (dry area). MATERIAL AND METHODS: Both physicochemical characteristics (humidity, water activity, Brix, ash content, pH, and total titratable acidity) and biochemical characteristics (total carotenoid content, betalain content, total polyphenolic content, and ascorbic acid content) were were studied according to previously reported methods. RESULTS: Regarding the physiochemical criteria, the moisture of the fresh peels of studied varieties ranged from 81.59 ± 0.02 to 83.47 ± 0.02%. The water activity (aw) ranged from 0.862 ± 0.001 to 0.872 ± 0.001. The values of Brix varied from 14.69 ± 0.05° Bx to 15.80 ± 0.03° Bx. pH values varied from 5.13 ± 0.01 to 5.32. The total titratable acidity values ranged from 0.130 ± 0.008 to 0.196 ± 0.014 g of citric acid/100 g of FM (fresh matter). The ash content values ranged from 8.92 ± 0.10 to 11.04 ± 0.06 g/100 g of FM. Regarding the biochemical criteria, the total carotenoid content ranged from 2.29 ± 0.01 to 2.87 ± 0.01 µg/g of FM. The total betalain content ranged from 6213.46 ± 58.86 to 8487.19 ± 51.71 µg/100 g of FM. The total polyphenolic content varied from 160 ± 3.55 to 243.79 ± 5.55 mg GA E/100 g of FM. The ascorbic content ranged from 58.21 ± 0.24 to 74.72 ± 0.17 mg/100 g of FM. CONCLUSION: The findings of physicochemical and biochemical criteria of the investigated varieties growing in Moroccan drylands showed promising results in terms of studied parameters.

Phytochemical Screening and Toxicological Study of Aristolochia baetica Linn Roots: Histopathological and Biochemical Evidence.

Aristolochia baetica (A. baetica) is a wild species of Aristolochiaceae family; its roots are used by Moroccan people against cancer for many years ago. The objective of the study was to investigate the phytochemical screening, acute and subacute toxicity of A. baetica roots growing in the north of Morocco. Qualitative and quantitative analyses of A. baetica roots were performed using standard methods; the acute toxicity of the root extract of the studied plant was assessed in mice by gavage of single doses of 1, 2, and 4 g/kg body weight for 14 days; by the time the subacute toxicity was done using repeated doses 1, 1.5, and 2 g/kg/day for 28 days. Histological changes and biochemical parameters as markers of kidney and liver function were evaluated. The results of phytochemical screening showed the presence of polyphenols, tannins, alkaloids, flavonoids, saponins, and the absence of anthraquinones, sterols, and terpenes. The results of acute toxicity showed the absence of mortality and signs of toxicity in groups treated with 1 and 2 g/kg; however, the clinical signs of toxicity were important and the rate of mortality was estimated at 16 % in the group treated with 4 g/kg. The results of subacute toxicity showed several changes of serum parameters registered in groups treated with 1.5 and 2 g/kg/day, respectively. The results showed also the absence of histological injuries in groups treated with 1 and 1.5 g/kg/day; meanwhile, the histological alterations were remarkable in treated group with the highest dose administered of 2 g/kg/day. The outcome of this work showed that the roots' extract of the studied plant was toxic in mice with repeated doses, but no toxic effect was observed with a single dose under 4g/kg.

Phytochemical Analysis and Toxicity Study of Aristolochia paucinervis Rhizomes Decoction Used in Moroccan Alternative Medicine: Histopathological and Biochemical Profiles.

ETHNOPHARMACOLOGICAL RELEVANCE: Aristolochia paucinervis (A. paucinervis) (Aristolochiaceae) is a plant frequently used in Moroccan alternative medicine. The aim of the current study is to investigate the phytochemical composition of rhizomes decoction of A. paucinervis (RDA) and to evaluate its acute and subacute toxicity following the OECD guidelines. MATERIALS AND METHODS: The qualitative phytochemical analysis of A. paucinervis was performed using standard qualitative phytochemical procedures. The acute toxicity of rhizomes decoction of the studied plant was evaluated in mice at single doses of 1, 2, and 4 g/kg of body weight for 14 days. In subacute toxicity study, the decoction was orally administered to mice at three different doses (0.5, 1, and 1.5 g/kg/day) for 28 days. Histopathological and biochemical parameters were investigated. RESULTS: The preliminary phytochemical screening showed the presence of flavonoids, saponins, alkaloids, and polyphenols and the absence of anthraquinones, sterols, and terpenes. There was no mortality and no significant changes occurred in animals treated with 1 and 2 g/kg in the acute toxicity model. The signs of toxicity and morbidity were remarkable with the highest tested dose (4g/kg). LD50 (dose required to kill 50% of the test population) was determined as 4 g/kg. Repeated oral administration of 1 and 1.5 g/kg/day of RDA for 28 days induced significant disturbance of serum parameters (AST, ALT, LDH, urea, creatinine). Kidney and liver extracted from mice fed with 1 and 1.5 g/kg/day showed significant histopathological injuries as tubular necrosis, inflammatory infiltrate, tubular degeneration, necrosis, and hepatic cholestasis. Meanwhile, neither histopathological nor biochemical alterations were observed in mice treated with 0.5 g/kg/day of body weight in comparison to the control group. CONCLUSION: RDA showed toxicity in mice at a dose of 1 g/kg/day under subacute toxicity conditions. RDA is safe at a single dose inferior to 4 g/kg of body weight. The plant extract prepared by decoction showed more poisonous effect than the extract prepared by maceration at room temperature.

Antioxidant and Antiproliferative Activities of Bioactive Compounds Contained in Rosmarinus officinalis Used in the Mediterranean Diet.

BACKGROUND: Rosmarinus officinalis (R. officinalis) is a medicinal plant called rosemary, largely used in the Mediterranean diet for many decades ago. OBJECTIVE: The aim of the present study was to investigate the polyphenolic content, the antioxidant activity, and the antiproliferative effect against human prostate cancer cell lines (LNCaP) of carnosol and carnosic acid as bioactive compounds contained in R. officinalis growing in Morocco. MATERIALS AND METHODS: Polyphenolic content of R. officinalis ethanolic extract was studied using colorimetric assay. Carnosol and carnosic acid contained in R. officinalis extract were quantified using high-performance liquid chromatography (HPLC). The antiproliferative effect of the studied extracts on LNCaP was evaluated by WST-1 bioassay, and the antioxidant activity was assessed using DPPH assay. RESULTS: The extracts of R. officinalis showed an important polyphenolic content ranging from 74.15 µg·GAE/mg to 146.63 µg·GAE/mg. The percentage of carnosol and carnosic acid in rosemary crops ranges from 11.7 to 17.3% and 1.09% to 3%, respectively. The extracts of R. officinalis exhibited a promoting antioxidant activity with IC50 ranging from 0.236 mg/mL to 0.176 mg/mL. Regarding the antiproliferative effect, the WST-1 assay revealed that all the tested extracts reduced notably the cell viability with IC50 values ranging from 14.15 to 15. 04 µg/mL. CONCLUSION: In the current work, carnosol and carnosic acid exhibit antioxidant and antiproliferative activities in a concentration-dependent manner.