Investigation on colorectal cancer and human herpesvirus infection among Algerian patients.

INTRODUCTION: Herpesviruses are a widespread family of double-stranded DNA viruses that establish life-long persistent infection in their hosts. Cumulative evidence tends to argue for the association of human herpesviruses, such as Kaposi's sarcoma herpesvirus (KHSV), Epstein-Barr virus (EBV), and human cytomegalovirus (HCMV) with various human disorders and diseases. The present study aims to investigate the presence of herpesviruses in colorectal cancer (CRC). METHODOLOGY: We investigated the presence of herpesviruses in 69 formalin-fixed paraffin embedded tissue (FFPE) biopsies, using a pan-herpesvirus nested polymerase chain reaction (PCR) with degenerate primers and HCMV specific primers to identify the presence of herpesviruses in CRC tissue. RESULTS: None of the samples we examined were positive for herpesviruses. CONCLUSIONS: Our results suggest that there is no (or very low) prevalence of lifelong herpesvirus infection in Algerian CRC patients. Larger cohorts may provide more insight into the prevalence of herpesviruses in Algerian CRC biopsies.

Phenolic profile, safety, antioxidant and anti-inflammatory activities of wasted Bunium ferulaceum Sm. aerial parts.

The pharmaceutical and nutraceutical industries benefit greatly from recycling and transforming non-utilized parts of medicinal plants from agro-industrial operations into value added products. Hence, the aim of this work was to study the potential nutraceutical and pharmaceutical applications of Bunium ferulaceum Sm. aerial parts, in order to maximize their value. The phenolic profile of their hydromethanolic extract was determined and its antioxidant activity was evaluated in vitro and in vivo alongside with its anti-inflammatory activity and safety profile. The extract exerted an in vitro antioxidant activity mainly through radical scavenging (DPPH IC50: 14.0 ± 0.3 µg/ml) and iron chelating ability (24 ± 2 µg/ml), while, in vivo, the extract did not cause any mortality or visible signs of acute toxicity at high dose (2000 mg/kg body weight). The supplementation of the extract at different doses improved mice liver redox state by increasing catalase and reduced glutathione levels and reducing lipid peroxidation, without causing any toxicity. Moreover, the extract efficiently inhibited xylene induced ear inflammation (62 %). These different bioactivities were linked to the phenolic compounds present in the extract, particularly, chlorogenic acid (78 ± 6 mg/g extract), rutin (44 ± 2 mg/g extract) and hesperidin (56 ± 9 mg/g extract). However, further studies should be carried out on the isolated major compounds found in the extract to correlate the activity with these compounds or their mixture. The wasted aerial parts of Bunium ferulaceum Sm. proved to be a valuable source of polyphenols and exhibited interesting health promoting effects with no toxicity. Thus, Bunium ferulaceum Sm. aerial parts can be included in nutraceutical formulations or used as functional food and the extracted compounds may be used as an alternative food preservative.

Inula viscosa phenolic extract suppresses colon cancer cell proliferation and ulcerative colitis by modulating oxidative stress biomarkers.

Inula viscosa is a perennial herbaceous plant native to the Mediterranean Basin, which is used topically for the treatment of various diseases in folk medicine. This study aimed to evaluate the in vivo intestinal anti-inflammatory activity of the ethanolic extract of I. viscosa (EEIV) and to test its effect on a colorectal cancer cell line. EEIV was administered to rats orally and daily at 100 and 200 mg/kg body weight for 7 days, and then colitis was induced by intrarectal instillation of 2 ml of 4% (v/v) acetic acid (AA) solution. At the end of the experiment, clinical examinations of the rats were conducted by evaluating macroscopic and histological signs of colonic tissues and measuring erythrocyte sedimentation rate (ESR) and the levels of C-reactive protein, fibrinogen, myeloperoxidase (MPO), malondialdehyde (MDA) and nitric oxide (NO). Using MTS assay, the antiproliferative effect of EEIV against human colon carcinoma HT29 cells and cytotoxicity on nondifferentiated Caco-2 cell line was evaluated. EEIV significantly decreased the ESR and fibrinogen levels as compared to control colitic rats (P < 0.001). It also significantly decreased the NO, MDA, and MPO levels in the colon tissue compared with the untreated colitic group (P < 0.001). These results were confirmed by macroscopic and histological examination, which showed significant protection against AA-induced ulcerative colitis. Furthermore, EEIV at a concentration of 369.88 µg/ml did not show cytotoxicity on confluent Caco-2 cells, with significant inhibition of colorectal cancer cell (HT29) growth (EC50 = 62.39 µg/ml). These results demonstrate that EEIV plays a potential role as a pharmacological tool in the management of inflammatory bowel disease and prevention of colorectal cancer.

Formulation and characterization of zein/gum arabic nanoparticles for the encapsulation of a rutin-rich extract from Ruta chalepensis L.

Rutin, a plant flavonol characterized by a wide range of biological effects, has limited application in foods because of its low water solubility and scarce bioavailability. This work aimed to investigate the encapsulation of a rutin-rich extract (200.6 ±â€¯1.5 mg/g of rutin) from Ruta chalepensis L. in zein nanoparticles (hydrodynamic diameter of 80-170 nm) prepared by antisolvent precipitation and stabilized by gum arabic (GA). The addition of GA (1:1 mass ratio with zein) significantly reduced the instability phenomena of zein nanoparticles through the deposition of a negatively charged layer as evidenced by the zeta potential and the UV-visible measurement, suggesting an electrostatic interaction between zein and GA. It also contributed to enhancing the encapsulation efficiency of rutin and inducing a rapid release during simulated digestion. These findings show that zein/GA nanoparticles represent a promising delivery system for natural extracts, fabricated through a facile and versatile process.

Anti-inflammatory activity of ethyl acetate and n-butanol extracts from Ranunculus macrophyllus Desf. and their phenolic profile.

ETHNOPHARMACOLOGICAL RELEVANCE: The members of the genus Ranunculus have counter-irritating properties and thus, they are traditionally used for treating anti-inflammatory disorders and other skin conditions. Ranunculus macrophyllus Desf. is a wild medicinal plant growing in Algeria and traditionally used to treat some cutaneous skin disorders. AIM: The aim of this study was to characterize the composition of the ethyl acetate and n-butanol extracts from Ranunculus macrophyllus Desf. as well as to elucidate and to compare their effect against acute skin inflammation. Moreover, both the antioxidant activity and the acute toxicity of the plant extracts were also studied. MATERIALS AND METHODS: Spectrophotometric and chromatographic methods were employed to identify and quantify phenolic compounds and triterpenoids from R. macrophyllus Desf. fractions. The antioxidant activity was estimated using the phosphomolebdenum, DPPH, reducing power and ß-carotene bleaching assays. The ethyl acetate and n-butanol extracts were screened for their anti-inflammatory activities using ex-vivo membrane stabilizing assays and in-vivo acute skin inflammation model. RESULTS: Ethyl acetate fraction showed the highest amounts of total phenolic compounds (413 ± 4 µg GAE/mg extract) and triterpenoids (70.4 ± 1.8 µg UAE/mg extract). Rutin, hesperidin, myricetin and kaempferol were the major compounds identified in the different fractions. Ethyl acetate fraction exhibited strong DPPH• radical scavenging ability (IC50 1.6 ± 0.2 µg/mL), high total antioxidant capacity (447 ± 7 µg AAE/mg extract) and reducing power (514 ± 8 µg AAE/mg extract). Ethyl acetate fraction inhibited (73.4 ± 0.3) % of linoleic acid peroxidation. Ethyl acetate and n-butanol fractions did not have any visible toxicity at 2000 mg/kg and presented excellent membrane stabilizing ability. The inhibition of xylene induced ear inflammation was (38 ± 4) % and (46 ± 1) % for RM-B and RM-EA, respectively. CONCLUSIONS: The high content of both phenolic compounds and triterpenoids combined with the remarkable anti-inflammatory effect and antioxidant activity of ethyl acetate and n-butanol extracts from R. macrophyllus Desf. support the wide spread use of this traditional plant on some skin disorders (inflammatory skin disorders).

Paronychia argentea Lam. protects renal endothelial cells against oxidative injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Paronychia argentea Lam. (Arabic tea), a species spontaneously growing in the Mediterranean area, has been used in folk medicine for renal diseases. AIM OF THE STUDY: To assess the antioxidant and protective potentials of different extracts from P. argentea in the renal endothelial NRK-52E cell line by several in vitro models, including a H2O2-induced oxidative stress model. MATERIAL AND METHODS: Aerial parts of P. argentea were collected in Algeria and ethanolic, chloroform and aqueous-chloroform extracts were obtained from dried plant. The antioxidant capacity was first evaluated by the Oxygen Radical Absorbance Capacity (ORAC) and the free radical scavenging activity (DPPH) methods. Cellular viability was assessed by MTT method assay after 24 h pretreatment with each extract concentration in order to measure protection from H2O2 in NRK-52E cells. Furthermore, the intracellular ROS formation (DCFH-DA method), was determined. RESULTS: P. argentea showed in vitro antioxidant activity as evidenced by the ORAC and DPPH assays. No cell toxicity was observed for concentrations ranging from 0.1 to 100 µg/mL of each extract. These extracts also exerted a protective effect on renal endothelial cells simultaneously treated with 1 mM H2O2. Chemical composition for the aqueous-chloroform extract was assessed by HPLC, as it showed the strongest antioxidant ability, revealing three quercetin derivatives as the main phenolic compounds. CONCLUSION: P. argentea is endorsed with antioxidant activity and protects renal endothelial cells against oxidative damage which indicate this plant constitutes a potential treatment for renal diseases.

Nortropane alkaloids as pharmacological chaperones in the rescue of equine adipose-derived mesenchymal stromal stem cells affected by metabolic syndrome through mitochondrial potentiation, endoplasmic reticulum stress mitigation and insulin resistance alleviation.

OBJECTIVES: Equine metabolic syndrome (EMS) refers to a cluster of associated abnormalities and metabolic disorders, including insulin resistance and adiposity. The numerous biological properties of mesenchymal stem cells (MSCs), including self-renewal and multipotency, have been the subject of many in-depth studies, for the management of EMS; however, it has been shown that this cell type may be affected by the condition, impairing thus seriously their therapeutic potential. Therefore, an attempt to rescue EMS adipose-derived stem cells (ASCs) with calystegines (polyhydroxylated alkaloids) that are endowed with strong antioxidant and antidiabetic abilities was performed. METHODS: ASCs isolated from EMS horses were subsequently treated with various concentrations of total calystegines. Different parameters were then assessed using flow cytometry, confocal as well as SE microscopy, and RT-qPCR. RESULTS: Our results clearly demonstrated that calystegines could improve EqASC viability and proliferation and significantly reduce apoptosis, via improvement of mitochondrial potentiation and functionality, regulation of pro- and anti-apoptotic pathways, and suppression of ER stress. Furthermore, nortropanes positively upregulated GLUT4 and IRS transcripts, indicating a possible sensitizing or mimetic effect to insulin. Most interesting finding in this investigation lies in the modulatory effect of autophagy, a process that allows the maintenance of cellular homeostasis; calystegines acted as pharmacological chaperones to promote cell survival. CONCLUSION: Obtained data open new perspectives in the development of new drugs, which may improve the metabolic dynamics of cells challenged by MS.

Characterization by liquid chromatography-mass spectrometry and antioxidant activity of an ethanolic extract of Inula viscosa leaves.

Inula viscosa (I. viscosa) is a common Mediterranean plant, well known for its content on bioactive molecules. The chemical composition of an ethanolic extract from I. viscosa leaves, growing in Algeria, was analysed by liquid chromatography coupled to photodiode array detection and electrospray ionization mass spectrometry (LC-DAD-ESI-MS/MS) operating in negative and positive mode. The methodology used revealed the presence of 51 compounds from which 47 were putatively identified, including 11 phenolic acids, 23flavonoids, one lignan and 12 terpenoids. Twenty-six of these compounds are described for the first time in I. viscosa. Antioxidant activity was measured using three different and complementary chemical assays: DPPH radical scavenging activity, oxygen radical absorbance capacity (ORAC) and hydroxyl radical scavenging capacity (HOSC). Results demonstrate that ethanolic leaf extract exhibit a high scavenging ability against DPPH (157.72 ±â€¯6.45 µM TE/g DW), peroxyl (4471.42 ±â€¯113.16 µM TE/g DW) and hydroxyl (630.10 ±â€¯17.81 µM TE/g DW) radicals, indicating that I. viscosa can be a promising source of bioactive compounds.

Evaluation of antidiabetic effect of total calystegines extracted from Hyoscyamus albus.

BACKGROUND: Hyoscyamus albus L. (Solanaceae) an old medicinal plant is a rich source of tropane and nortropane alkaloids which confers to this plant a number of very interesting and beneficial therapeutic effects. PURPOSE: Calystegines that are polyhydroxylated alkaloids and imino-sugars poccess significant glycosidases inhibitory activities and are therefore good candidats for the treatment of diabetes mellitus. STUDY DESIGN: Calystegines extracted from Hyoscyamys albus seeds were tested for teir acute oral toxicity and investigated for their in-vivo antidiabetic effect on Streptozotocine induced diabetes in mice. METHODES: Calystegines were extracted from the seeds plant using an Ion exchange column; the remaining extract was then administrated orally to mice at several single doses for acute toxicity assay. A dose of 130mg/kg streptozotocine was injected to mice to induce diabetes mellitus, and diabetic mice were treated orally during 20days with 10mg/kg and 20mg/kg calystegines and 20mg/kg glibenclamide as the reference drug. RESULTS: Acute oral toxicity showed that calystegines are not toxic up to a dose of 2000mg/kg with absence of any signs of intoxication and damages in Liver and kidney tissues. The nortropane alkaloids markedly reduced blood glucose levels and lipid parameters of diabetic mice to normal concentrations after 20days of treatment at 10mg/kg and 20mg/kg (p<0.05). Histopathological study of diabetic mice pancreas indicated that calystegines of Hyoscyamus albus have minimized streptozotocine damages on ß-cells of islets of langerhans, stimulated ß-cells regeneration and improved with this insulin secretion. CONCLUSION: The findings of this study suggest that calystegines are potent antidiabetic agents with antihyperglicemic and hypolipidemic effects, and a protective fonction on pancreas in streptozotocin induced diabetes in mice.

Inhibition of myeloperoxidase activity by the alkaloids of Peganum harmala L. (Zygophyllaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Seeds and aerial parts of Peganum harmala L. are widely used in Algeria as anti-inflammatory remedies. Evaluation of Peganum harmala total alkaloids extracts and pure ß-carboline compounds as an anti-inflammatory treatment by the inhibition of an enzyme key of inflammatory, myeloperoxidase (MPO) and HPLC quantification of the alkaloids from the different parts of plant. MATERIALS AND METHODS: MPO inhibition was tested using taurine chloramine test. The inhibition of LDL oxidation induced by MPO was carried out. The molecular docking analysis of Peganum harmala alkaloids on MPO was performed using the Glide XP docking protocol and scoring function and the redox potential of alkaloids was determined using an Epsilon potentiostat. The concentration of harmala alkaloids was determined using HPLC analysis. RESULTS: The HPLC profiling of the active total alkaloids indicates that ß-carboline e.g. harmine, harmaline, harmane, harmol and harmalol are major components. As ß-carbolines resemble tryptamine, of which derivatives are efficient inhibitors of MPO, the harmala alkaloids were tested for their activity on this enzyme. Total alkaloids of the seeds and of the aerial parts strongly inhibited MPO at 20µg/mL (97±5% and 43±4%, respectively) whereas, at the same concentration, those of the roots showed very low inhibition (15±6%). Harmine, harmaline and harmane demonstrated a significant inhibition of MPO at IC50 of 0.26, 0.08 and 0.72µM respectively. These alkaloids exerted a similar inhibition effects on MPO-induced LDL oxidation. Molecular docking analysis of Peganum harmala alkaloids on MPO showed that all active Peganum harmala alkaloids have a high affinity on the active site of MPO (predicted free energies of binding up to -3.1kcal/mol). Measurement of redox potentials versus the normal hydrogen electrode clearly differentiated (i) the high MPO inhibitory activity of harmine, harmaline and harmane (+1014, 1014 and 1003mV, respectively); and (ii) the low activity of harmalol and harmol (+629/778 and 532/644mV, respectively). A reverse phase HPLC method has been developed to determine simultaneously five alkaloids of Peganum harmala. Seeds contained all five ß-carboline derivatives with the main active alkaloids, harmaline and harmine, being up to 3.8% and 2.9%, respectively. Up to 3.2% of harmine was determined in the roots. The four ß-carboline derivatives, harmine, harmaline, harmane and harmalol were identified in the aerial parts. The highest inhibitory effect observed in seeds and the moderate effect of aerial parts could be explained by their harmine and harmaline content. In contrast, the very weak inhibition of the root extract, despite the presence of harmine, may tentatively be explained by the high concentration of harmol which can reduce Compound II of MPO to the native form. CONCLUSION: The inhibition of MPO by Peganum harmala ß-carboline alkaloids, herein reported for the first time, may explain the anti-inflammatory effect traditionally attributed to its herbal medicine.

Identification and quantification of the main active anticancer alkaloids from the root of Glaucium flavum.

Glaucium flavum is used in Algerian folk medicine to remove warts (benign tumors). Its local appellations are Cheqiq el-asfar and Qarn el-djedyane. We have recently reported the anti-tumoral activity of Glaucium flavum root alkaloid extract against human cancer cells, in vitro and in vivo. The principal identified alkaloid in the extract was protopine. This study aims to determine which component(s) of Glaucium flavum root extract might possess potent antitumor activity on human cancer cells. Quantitative estimation of Glaucium flavum alkaloids was realized by HPLC-DAD. Glaucium flavum effect on human normal and cancer cell viability was determined using WST-1 assay. Quantification of alkaloids in Glaucium flavum revealed that the dried root part contained 0.84% of protopine and 0.07% of bocconoline (w/w), while the dried aerial part contained only 0.08% of protopine, glaucine as the main alkaloid, and no bocconoline. In vitro evaluation of the growth inhibitory activity on breast cancer and normal cells demonstrated that purified protopine did not reproduce the full cytotoxic activity of the alkaloid root extract on cancer cell lines. On the other hand, bocconoline inhibited strongly the viability of cancer cells with an IC50 of 7.8 µM and only a low cytotoxic effect was observed against normal human cells. Our results showed for the first time that protopine is the major root alkaloid of Glaucium flavum. Finally, we are the first to demonstrate a specific anticancer effect of Glaucium flavum root extract against breast cancer cells, which can be attributed, at least in part, to bocconoline.

Revealing the anti-tumoral effect of Algerian Glaucium flavum roots against human cancer cells.

Glaucium flavum (G. flavum) is a plant from the Papaveraceae family native to Algeria where it is used in local traditional medicine to treat warts. G. flavum root crude alkaloid extract inhibited breast cancer cell proliferation and induced G2/M phase cycle arrest and apoptosis without affecting normal cells, which is a highly awaited feature of potential anti-cancer agents. G. flavum significantly reduced growth and vascularization of human glioma tumors on chicken chorioallantoic membrane (CAM) in vivo. The chromatographic profile of the dichloromethane extract of G. flavum root showed the presence of different constituents including the isoquinoline alkaloid protopine, as the major compound. We report for the first time that G. flavum extract may represent a new promising agent for cancer chemotherapy.