Close relationship between high HbA1c levels and methemoglobin generation in human erythrocytes: The enhancement of oxidative stress in the process.

OBJECTIVE: This study aimed to investigate the effect of diabetic plasma on human red blood cells (RBCs) in order to highlight the amplification mechanisms of oxidative stress (OS) in relation to methemoglobin (metHb) production, a potential bio-indicator that could be related to diabetes disease. RESEARCH DESIGN AND METHODS: Normal RBCs were co-incubated with the diabetic plasma of 24 patients at different HbA1c levels, for 0, 24, and 48 h in order to assess cell turbidity and hemoglobin (Hb) stability. Hb and metHb production were quantified inside and outside RBCs. Malonaldehyde (MDA) level and cell morphology were concomitantly evaluated. RESULTS: The cell turbidity was significantly decreased in the group co-incubated with diabetic plasma at high HbA1c levels (0.074 ± 0.010 AU) compared to the control group (0.446 ± 0.019 AU). A significant decrease in intracellular Hb (0.390 ± 0.075 AU) and its stability (0.600 ± 0.001 AU) were revealed. Also, we found an important increase of metHb levels inside RBCs (0.186 ± 0.017 AU) and in its supernatant (0.086 ± 0.020 AU) after 48 h. Consequently, MDA absorbance increased significantly (0.320 ± 0.040 AU) in RBCs exposed to diabetic plasma with high HbA1c. CONCLUSION: These findings suggest that poor glycemic control in diabetes leads to metHb generation which is the main factor of the OS amplification.

The Effect of In Ovo Administration of Rosemary Essential Oil on Hatchability, Relative Hatching Weight, and Embryo Mortality Rate in Japanese Quail (Coturnix coturnix japonica).

This study aimed to determine the effects of air sac injection of rosemary essential oil at different concentrations in ovo in quail eggs on hatching rate, relative chick weight at hatching, and embryonic mortality rate. A total of 1060 Japanese quail eggs were divided into four groups: negative control (non-injected), positive control (30 µL sterile distilled water/egg), and two treated groups with 1 and 3 µL oil/egg, respectively. The concentration of 3 µL/egg showed a toxic effect on embryonic development, as revealed by the significantly (p = 0.015) higher post-injection mortality rate (18.21%) compared to 1 µL/egg with 8.3%. Furthermore, hatchability was significantly increased (p = 0.0001) with 1 µL/egg compared to 3 µL/egg with 69.1% and 44.48%, respectively. No significant difference was observed between the concentration of 1 µL/egg and the control groups (p = 0.822). Both l and 3 µL essential oil/egg significantly enhanced (p = 0.0001) relative chick weight at hatching by 67.14% and 70.32%, respectively, compared to the control groups. In conclusion, injecting eggs with 1 µL oil/egg showed positive effects both on hatching and relative chick weight. The concentration of 3 µL/egg was revealed to be toxic, with dramatic effects on embryonic survival.

First Descriptive Analysis of the Faecal Microbiota of Wild and Anthropized Barbary Macaques (Macaca sylvanus) in the Region of Bejaia, Northeast Algeria.

Previous research has revealed the gut microbiota profile of several primate species, as well as the impact of a variety of anthropogenic factors, such as tourist food supply, on these bacterial communities. However, there is no information on the gut microbiota of the endangered wild Barbary macaque (Macaca sylvanus). The present study is the first to characterize the faecal microbiota of this species, as well as to investigate the impact of tourist food provisioning on it. A total of 12 faecal samples were collected in two groups of M. sylvanus in the region of Bejaia in Algeria. The first group-a tourist-provisioned one-was located in the tourist area of the Gouraya National Park and the second group-a wild-feeding one-was located in the proximity of the village of Mezouara in the forest of Akfadou. After DNA extraction, the faecal microbiota composition was analysed using 16S rDNA sequencing. Statistical tests were performed to compare alpha diversity and beta diversity between the two groups. Non-metric multidimensional scaling analysis (NMDS) was applied to visualize biodiversity between groups. Behaviour monitoring was also conducted to assess the time allocated to the consumption of anthropogenic food by the tourist-provisioned group. Our results revealed the presence of 209 bacterial genera from 17 phyla in the faecal microbiota of Barbary macaques. Firmicutes was the most abundant bacterial phylum, followed by Bacteroidetes and Verrucomicrobia. On the other hand, the comparison between the faecal microbiota of the two study groups showed that tourism activity was associated with a significant change on the faecal microbiota of M.sylvanus, probably due to diet alteration (with 60% of feeding time allocated to the consumption of anthropogenic food). The potentially low-fibre diet at the tourist site adversely influenced the proliferation of bacterial genera found in abundance in the wild group such as Ruminococcaceae. Such an alteration of the faecal microbiota can have negative impacts on the health status of these animals by increasing the risk of obesity and illness and calls for special management measures to reduce the provisioning rate in tourist areas.

Oxidative stress as an underlying mechanism of anticancer drugs cytotoxicity on human red blood cells' membrane.

The aim of this study is to investigate the direct in vitro effects of anticancer drugs on red blood cells (RBCs) and to explore the underlying mechanism, mainly by measuring RBCs oxidative stress (OS) status. After RBCs direct contact with fourteen (14) anticancer drugs, several parameters were assessed including: cellular turbidity, methemoglobin (metHb) generation, released Hb and Hb stability. Moreover, intracellular Hb, considered as new molecular target of anticancer drugs, was quantified inside RBCs. MDA level, the main biomarker of OS, was simultaneously measured. The cellular turbidity reveled severe (docetaxel "TXT", 0.03 ± 0.002), moderate (methotrexate "MTX", 0.49 ± 0.009), or none (5-fluorouracil "5-FU", 0.76 ± 0.029) membrane cytotoxicity (MC). An inverse relationship between cell concentration, released Hb and metHb content was obtained. High metHb generation, revealing intense OS, was also mostly expressed in paclitaxel "TXL" and etoposide "VP16". Further, epirubicin "EPI" and "TXT" induced important oxidation of membrane lipids with 0.32 ± 0.014 and 0.26 ± 0.004, respectively. Also, MTX (0.17 ± 0.006) and doxorubicin "DOX" (0.32 ± 0.034) affected significantly Hb stability by a direct contact with molecule. These findings demonstrated that anticancer drugs have the ability to induce membrane damages by the exacerbation of OS through membrane lipid peroxidation and Hb oxidation even inside RBCs.

The Use of Cyclodextrin or its Complexes as a Potential Treatment Against the 2019 Novel Coronavirus: A Mini-Review.

Severe Acute Respiratory Syndrome Coronavirus 2 has spread rapidly since its discovery in December 2019 in the Chinese province of Hubei, reaching this day all the continents. This scourge is, unfortunately, in lineage with various dangerous outbreaks such as Ebola, Cholera, Spanish flu, American seasonal flu. Until today, the best solution for the moment remains prevention (Social distancing, hand disinfection, use of masks, partial or total sanitary containment, etc.); there is also the emergence of drug treatment (research and development, clinical trials, use on patients). Recent reviews emphasized the role of membrane lipids in the infectivity mechanism of SARS-COV-2. Cholesterol-rich parts of cell membranes serve as docking places of host cells for the viruses. Coronavirus 2 is a member of a virus family with lipid envelope that fuses with host cell through endocytosis, internalizing its components in the cell. In vitro cell models have shown that depletion of cholesterol by cyclodextrin, and particularly methyl beta cyclodextrin disturb the host cell membrane lipid composition this way, reducing the attachment of the virus to the protein receptors. This review aims to summarize the state of the art of research concerning the use of cyclodextrin or its complexes as a potential treatment against this new virus and update work already published.

New Formulation and Evaluation of Camptothecin Encapsulated and/or Dispersed Suppository.

BACKGROUND: Camptothecin is known for its potent anticancer activity. However, its optimal activity is reduced due to its low solubility and stability in biological media. OBJECTIVE: The aim of the present study is to design and characterize a Camptothecin (CPT) suppository formulation. METHODS: Rectal suppositories of camptothecin alone, encapsulated with Cyclodextrin (CD) and in the ternary system (CPT encapsulated with cyclodextrin and dispersed in Polyethylene Glycol (PEG) 6000) were prepared using various hydrophobic and hydrophilic polymeric bases as semi-synthetic glyceride (Suppocire® AM Pellets) and Polyethylene Glycols (PEGs) mixtures. Formulations were evaluated by various parameters like weight variation, drug content, hardness and liquefaction time. In vitro release study was performed in USP type I apparatus using phosphate buffer pH 7.2 as dissolution media. RESULTS: Suppositories were within the permissible range of all physical parameters. In vitro drug released from water soluble base (PEG) was greater than that from oil soluble base with ninety percent (90%) of drug dissolution. It was also established that drug release from various formulations was by diffusion mechanism, according to the Higuchi's equation. CONCLUSION: This new formulation offers a new approach to colorectal cancer treatment by offering an alternative and simple drug administration route.

High HbA1c levels affect motility parameters and overexpress oxidative stress of human mature spermatozoa.

The aim of this study is to investigate, by a validated in vitro model, the effect of diabetic plasma on ejaculated human spermatozoa. Plasma of 51 male diabetic patients (mean age 62.28 ± 9.28 years) was selected according to their HbA1c level: low HBA1c  ≤ 5% (31 mmol/mol), moderate HBA1c 6%-8% (42-64 mmol/mol) and high HBA1c  ≥ 10% (86 mmol/mol). The plasma was tested on eighteen normal semen samples by analysing gametes motility using a computer Sperm Class Analyzer® and their corresponding oxidative stress (OS) status using thiobarbituric acid-reactive substances assay. The results indicated that diabetic plasma affected all sperm motility parameters with high HbA1c showing the most important deleterious effects. Low gametes' straight-line velocity was observed in high HbA1c level, mainly after 20 min of co-incubation (8.78 ± 0.47 µm/s). Also, the highest lipid peroxidation (nmoles MDA/108 SPZ) was observed in high HbA1c values (0.92 ± 0.09), higher than those in spermatozoa treated with H2 O2 (0.85 ± 0.04). Conclusively, a direct impact of diabetic plasma on spermatozoa is revealed with overexpression of OS as the underlying mechanism. These findings suggested that it is strongly recommended to control clinically the glycaemic level and OS in diabetic patients for the maintenance of male fertility.

Development and validation of HPLC method for simultaneous quantification of alpha-tocopherol (free or encapsulated) and cholesterol in semen cryopreservation media.

The HPLC method was developed and validated for assaying alpha-tocopherol and cholesterol in cryopreservation media. Chromatographic separation was performed on an isocratic system, using a C-18 column. The mobile phase was composed of a mixture of methanol:acetonitrile:water 68:28:4 (v/v/v), using a flow rate of 1.5 mL/min and 20 µL injection volume, at a wavelength of 208 nm. The method was validated according to International Conference on Harmonization guidelines. The method proved to be specific, accurate, precise, and linear with correlation coefficients greater than 0.996 over a wide concentration range of both analytes. Vitamin E and cholesterol presented limits of detection of 0.002 mg/mL, 0.026 mg/mL and limits of quantitation of 0.006 mg/mL, 0.086 mg/mL, respectively. This method is simple and rapid, shows high precision and accuracy, and offers the advantage of simultaneous assaying of vitamin E and cholesterol (alone, in cyclodextrins complexes or in liposome loaded) on semen cryopreservation media.

Effects of olive oil dietary supplementation on sperm quality and seminal biochemical parameters in rooster.

The current study investigated the effect of olive oil (OO) dietary supplementation on rooster semen quality. Three groups of local breed roosters fifty fourth weeks old were randomly assigned to either one of the following diets during 8 weeks: ad libitum standard diet without OO dietary supplementation (Group 1), ad libitum standard diet with 0.2 ml of daily OO supplementation per rooster (Group 2), and ad libitum standard diet with 0.4 ml of daily OO supplementation per rooster (Group 3). Semen quality (volume, concentration, viability and motility) and biochemical seminal parameters (glucose, cholesterol, triglycerides, HDL and LDL) were determined twice weekly from the 5th to 8th week of treatment. Results showed no significant difference between the three groups during the three first weeks for sperm concentration and sperm motility; at the fourth week, males provided with 0.4 ml OO (Group 3) showed a significantly improved sperm concentration, sperm viability and sperm motility compared to the others groups. Even if group 3 showed slight elevated values for glucose, triglycerides and LDL, no significant difference was observed for semen volume and biochemical seminal parameters between the three studied groups. The current results suggest that olive oil dietary supplementation could be an interesting alternative to enhance rooster sperm quality particularly in aged individuals where ejaculates quality declines systematically.

Characterization of the onset of puberty in Tazegzawt lambs, an endangered Algerian sheep: Body weight, thoracic perimeter, testicular growth, and seminal parameters.

AIM: The aim of the present study was to define the onset of puberty in Tazegzawt ram lambs, an Algerian sheep breed in endangered status with a small population in its local area. MATERIALS AND METHODS: Body growth (body weight and thoracic perimeter), scrotal circumference (SC), penis development stages, and seminal parameters (volume, concentration, and motility) were measured. Data were recorded at fortnightly intervals in 10 animals from 9 to 49 weeks of age. RESULTS: On the basis of seminal analyses, puberty occurred between 29 and 45 weeks of age. At 29 weeks of age, 30% of lambs reached puberty, and at 45 weeks of age, puberty was observed in 100% of the analyzed animals. Body weight appeared as the most determinant factor, and the onset of puberty was observed when animals reached 43.2±6.4 kg body weight with 25.8±3.7 cm of SC. Seminal analyses revealed that all parameters increased regularly from puberty onset except for sperm concentration. The mean semen volume during the study period was 0.48±0.33 mL with 0.84±0.6 mL at 37 weeks of age. Sperm concentration evolved similarly as semen volume; at 29 and 43 weeks of age, the sperm concentration was 942×106 and 1904×106 spermatozoa/mL, respectively. Kinematic parameters including the percentage of motility, the percentage of progressive motility, and gametes velocities as determined by Computer-Aided Sperm Analyzer showed the highest values at 49 weeks of age. CONCLUSION: The current results revealed that, in Tazegzawt ram lambs, puberty occurs between 29 and 45 weeks when animals reach 43.2±4.6 kg body weight.

Reproduction performance and blood biochemical parameters in dairy cows: Relationship with oxidative stress status.

BACKGROUND AND AIM: During the last decades, reproduction performances declined dramatically worldwide, but little is known concerning the involvement of oxidative stress as a causative factor. Oxidative stress may act at different levels, with negative impacts on cell membrane integrity and other active molecules with potential subsequent effects on reproduction. The aim of the current study was to investigate the oxidative stress status in cows according to their reproductive performances. MATERIALS AND METHODS: Peripheral blood concentration of two oxidative stress biomarkers, glutathione S-transferase (GST) and malondialdehyde (MDA), and other biochemical parameters (glucose, total lipids, cholesterol, triglycerides, albumin, total proteins, calcium, urea, creatinine, direct bilirubin, alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase) were determined in 40 healthy cows. Body condition score (BCS), calving to first service interval (FSI), calving to conception interval (CCI), and the number of service per conception (SPC) were simultaneously recorded for each cow. RESULTS: Concerning FSI, three groups were established: Group 1 (from 44 to 60 days), Group 2 (from 60 to 70 days), and Group 3 (from 70 to 80 days). For CCI, two groups were considered: Group 1 (<110 days) and Group 2 (>110 days). MDA showed significant high values only in cows with the lowest BCS (1.5) compared to cows with BCS note of 2.5 and 3.5. No significant difference was observed in cows oxidative stress status (MDA and GST) according to reproductive performances (FSI, CCI, and SPC) in all studied groups. CONCLUSION: The results revealed relatively altered oxidative stress status in cows with abnormal reproductive performances; however, no significant difference was recorded whatever the considered reproductive parameter.

Beneficial and harmful effects of cyclodextrin-vitamin E complex on cryopreserved ram sperm.

Vitamin E is a potent molecule, especially when loaded in cyclodextrin, in modulating oxidative stress during the freeze-thawing process. The present study aimed to investigate the effect of different concentrations of cyclodextrin-vitamin E complex (CD-Vit E) on cryopreserved ram sperm. Ejaculates collected from five adult rams were pooled and divided into four aliquots. All aliquots were treated in Tris-extender (Tris-glucose-citric acid) containing 2 mg cholesterol-loaded methyl-ß- cyclodextrin/120 × 106 spermatozoa and either 0 (Control), 2, 4 or 6 mg CD-Vit E/120 × 106 spermatozoa, corresponding to 0, 0.5, 1 or 1.5 of pure vitamin E, respectively. After incubation at 22 °C for 15 min and the addition of Tris-extender containing glycerol and egg yolk (v/v), all aliquots were frozen in liquid nitrogen. After thawing, motility (computer aided sperm analysis), viability (eosin staining), membrane integrity (HOST), acrosome integrity (Coomassie G-250 staining) and lipid peroxidation (Thiobarbituric acid assay) were evaluated. Compared to control, 2 mg CD-Vit E had a significant positive effect on total motility, progressive motility, movement linearity (LIN%), viability and lipid peroxidation. At 4 mg, however, CD-Vit E had a significant negative effect on total motility, progressive motility, membrane functionality and acrosome integrity. At a greater concentration (6 mg), the negative effects were greater as compared with inclusion of 4 mg in the cryoprotectant and the percentage of rapidly and moderately motile gametes and viability were also altered. In conclusion, the effect of CD-Vit E on cryopreserved ram sperm was concentration-dependent with the 2 mg amount having a beneficial effect while greater concentrations (4 and 6 mg) had a harmful effect on sperm motility and gamete integrity but without affecting oxidative stress status.

Male rat exposure to low dose of di(2-ethylhexyl) phthalate during pre-pubertal, pubertal and post-pubertal periods: Impact on sperm count, gonad histology and testosterone secretion.

Di (2-ethylhexyl) phthalate (DEHP) is the most ubiquitous endocrine disruptor in the environment. The present study aimed to investigate the low dose effects of DEHP on the male reproductive system of rats exposed during the pre-pubertal, pubertal and post-pubertal periods. Male Wistar rats were daily gavaged by DEHP from postnatal day (PND) 21 to PND 120 with 0.5, 50 and 5000µg/kgBW/d. A decrease in sperm count of 41%, 24% and 46% was observed at 0.5, 50 and 5000µg/kgBW/d respectively. A decrease of Sertoli cells number was observed at 50 and 5000µg/kgBW/d (22% and 42%, respectively). Non-monotonic dose-response was observed for testosterone levels with a significant increase at 50µg/kgBW/d associated to a notable enhancement of Leydig cells number (35%). In conclusion, our results showed that postnatal exposure to low doses of DEHP affects sperm count, Sertoli and Leydig cells number and testosterone level.

Elevated non-esterified fatty acid and ß-hydroxybutyrate in transition dairy cows and their association with reproductive performance and disorders: A meta-analysis.

A meta-analysis including 36 different results of statistic models from 14 papers was conducted. It evaluated the association between elevated non-esterified fatty acids and/or ß-hydroxybutyrate (BHB) on the reproduction outcomes that were pregnancy at first insemination, estrous cyclicity, time to pregnancy, metritis and placental retention. Each association between BHB or NEFA and an outcome reported in literature was a model considered as raw-data for the meta-regression. For each outcome, the meta-regression adjusted the odds ratio, relative risk or hazard ratio with various moderators to reduce the heterogeneity among the studies. The relative risk for metritis and placental retention in case of high BHB or NEFA was in accordance to previous meta-regression and was 1.91 (IC95 = 1.72 to 2.12) and 1.51 (95%CI = 1.19 to 1.92), respectively. The relative risk for pregnancy at first insemination in case of high BHB was assessed to be 0,62 (95%CI = 0,41 to 0,93). The hazard ratio for time to pregnancy in case of high BHB and NEFA was 0.77 (95%CI = 0.61 to 0.97). The present meta-analysis failed to clearly conclude on the association between estrus cyclicity and high BHB or NEFA. The present work allowed a new overview on the association between hyperketonemia and reproductive performance and disorders. It updated the previous meta-regression and included new outcomes. It highlighted the urgent need of further intensive epidemiologic studies on this topic.

Concurrent measurement of cellular turbidity and hemoglobin to evaluate the antioxidant activity of plants.

In past decades, a multitude of analytical methods for measuring antioxidant activity of plant extracts has been developed. However, when using methods to determine hemoglobin released from human erythrocytes treated with ginger extracts, we found hemoglobin concentrations were significantly higher than in untreated control samples. This suggests in the presence of antioxidants that measuring hemoglobin alone is not sufficient to determine hemolysis. We show concurrent measurement of erythrocyte concentration and hemoglobin is essential in such assays, and describe a new protocol based on simultaneous measurement of cellular turbidity and hemoglobin.

Extended spectrum ß-lactamase producing Escherichia coli in broiler breeding roosters: Presence in the reproductive tract and effect on sperm motility.

Extended spectrum ß-lactamases (ESBL)-producing Escherichia coli have emerged worldwide in animal husbandry and they were reported from different ecosystems. The purpose of this study was firstly, to investigate the presence of ESBL-producing E. coli in the gastrointestinal (GIT) and reproductive (RT) tracts of broiler breeding roosters, and secondly to study the impact of an ESBL-producing E. coli on artificially infected semen. A total of seventeen ESBL-producing E. coli strains were isolated from the gastrointestinal and reproductive tracts of nine broiler breeding roosters. All isolates were identified to the species level by API 20E system and MALDI-TOF, serotyped, and genetically characterized for ESBL production. Semen was artificially infected with E. coli ATCC25922 or with an ESBL-producing E. coli strain recovered from the reproductive tract. A computer aided semen analyzer (CASA) was used to compare different spermatozoa motility parameters in each sample. All ESBL-producing E. coli isolates could not be typed with the currently used sera and they were harboring a blaCTX-M gene alone or in combination with a blaTEM gene. The semen quality was notably less affected in samples infected with ESBL-producing E. coli strain compared to the control and sample infected with E. coli ATCC25922. The present study revealed that ESBL-producing E. coli can be isolated from both reproductive and digestive tracts of broiler breeding roosters. Contamination of the reproductive tract with ESBL-producing E. coli could lead to contamination of semen and could be an important factor in the dissemination of ESBL-producing E. coli in poultry.

In-vitro effects of Thymus munbyanus essential oil and thymol on human sperm motility and function.

Traditional medicine has been used worldwide for centuries to cure or prevent disease and for male or female contraception. Only a few studies have directly investigated the effects of herbal compounds on spermatozoa. In this study, essential oil from Thymus munbyanus was extracted and its effect on human spermatozoa in vitro was analysed. Gas chromatography and Gas chromatography-mass spectrometry analyses identified 64 components, accounting for 98.9% of the composition of the oil. The principal components were thymol (52.0%), γ-terpinene (11.0%), ρ-cymene (8.5%) and carvacrol (5.2%). Freshly ejaculated spermatozoa was exposed from control individuals to various doses of the essential oil for different time periods, and recorded the vitality, the mean motility, the movement characteristics (computer-aided sperm analysis), the morphology and the ability to undergo protein hyperphosphorylation and acrosomal reaction, which constitute two markers of sperm capacitation and fertilizing ability. In vitro, both the essential oil extracted from T. munbyanus and thymol, the principal compound present in this oil, impaired human sperm motility and its capacity to undergo hyperphosphorylation and acrosome reaction. These compounds may, therefore, be of interest in the field of reproductive biology, as potential anti-spermatic agents.

Amorphous solid dispersion studies of camptothecin-cyclodextrin inclusion complexes in PEG 6000.

Abstract: The present work focused on the solubility enhancement of the poorly water-soluble anti-cancer agent camptothecin which, in its natural state, presents poor solubility inducing lack of activity with a marked toxicity. A new approach is adopted by using a ternary system including camptothecin (CPT) and cyclodextrins (CDs) dispersed in polyethylene glycol (PEG) 6000. Camptothecin solubility variations in the presence of α-CD, ß-CD, γ-CD, hydroxypropyl-α-CD (HPα-CD), hydroxypropyl-ß-CD (HPß-CD), permethyl-ß-CD (PMß-CD) and sulfobutyl ether-ß-CD (SBEß-CD), were evaluated by Higuchi solubility experiments. In the second part, the most efficient camptothecin/P-CDs binary systems, mainly HPß-CD and PMß-CD, were dispersed in PEG 6000. In addition to a drug release and modeling evaluation, the CPT interactions with CDs and PEG 6000 to prepared the amorphous solid dispersion in the binary and ternary systems were investigated by Fourier transformed infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), thermogravimetric analyses (TGA) and X-ray powder diffraction (XRPD). The results showed that HPß-CD and PMß-CD were the most efficient for camptothecin solubilization with highest apparent equilibrium constants. Dissolution studies showed that percentage of CPT alone after two hour in 0.1 M HCI medium, did not exceed 16%, whereas under the same conditions, CPT/PMß-CD complex reached 76%. When dispersing the binary systems CPT/ß-CDs in PEG 6000, the velocity and the percentage of CPT release were considerably improved whatever the CD used, reaching the same value of 85%. The binary and ternary systems characterization demonstrated that CPT inclused into the CDs cavity, replacing the water molecules. Furthermore, a drug transition from crystalline to amorphous form was obtained when solid dispersion is realized. The present work demonstrated that ternary complexes are promising systems for CPT encapsulation, and offer opportunities to use non toxic and commonly solubilizing carriers: ßCD and PEG 6000 to improve bioavailability.

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.

Molecular mechanism underlying anti-inflammatory and anti-allergic activities of phytochemicals: an update.

The resort worldwide to edible medicinal plants for medical care has increased significantly during the last few years. Currently, there is a renewed interest in the search for new phytochemicals that could be developed as useful anti-inflammatory and anti-allergic agents to reduce the risk of many diseases. The activation of nuclear transcription factor-kappa B (NF-κB) has now been linked to a variety of inflammatory diseases, while data from numerous studies underline the importance of phytochemicals in inhibiting the pathway that activates this transcription factor. Moreover, the incidence of type I allergic disorders has been increasing worldwide, particularly, the hypersensitivity to food. Thus, a good number of plant products with anti-inflammatory and anti-allergic activity have been documented, but very few of these compounds have reached clinical use and there is scant scientific evidence that could explain their mode of action. Therefore, this paper intends to review the most salient recent reports on the anti-inflammatory and anti-allergic properties of phytochemicals and the molecular mechanisms underlying these properties.