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Objective The purpose of this study was to compare the antibacterial effect of two types of Eugenol against Streptococcus mutans and then assess the effect of different concentrations of two types of Eugenol on S. mutans adhesion on Nickel-Titanium (NiTi) orthodontic wires in comparison to in vivo wires with fluoride-based hygiene regimen. Materials and Methods Culture of Streptococcus mutans with NiTi orthodontic wires was done. Different media were prepared by adding 100%, 50%, 25%, and 12.5% of two origins of Eugenol (one biological obtained by hydrodistillation of Syzygium aromaticum and one chemical already prepared available in drugstores for dental use (Idental, MOROCCO, lot number: UAN/17–211/1). Three sizes of NiTi wires (0.016 inch, 0.016 × 0.022 inch, 0.017 × 0.025 inch) were retrieved from adult patients undergoing orthodontic treatment after 1 month of setting them up in the mouth. After incubation, colony forming unites were calculated and a SEM analysis was done to the surface of each wire. ANOVA test was done between all groups to find statistical differences and post-hoc t-test with Bonferroni analysis was performed to elucidate differences between all groups with α = 0.05. Results Eugenol has an anti-bacterial effect against S. mutans. The biological Eugenol has greater effect than the chemical one. The same observations were done for anti-adherent effect, the biological Eugenol demonstrated the highest anti-adherent effect at all concentrations while the effect of the chemical Eugenol was the lowest. Conclusions The origin and the extraction mode of Eugenol have a crucial role in its antimicrobial and anti-adherent effect. Eugenol might constitute an alternative to Fluoride because it has an anti-adherent effect, limiting the incidence of white spot lesions.

Aims: The purpose of this study was to assess the Density Gradient Centrifugation (DGC) effectiveness in spermatozoa selection by analyzing the oxidative stress profile and in pregnancy prediction outcomes of In Vitro Fertilization (IVF). Study Design: This was a prospective study. Place and Duration of Study: Department of Biology, laboratory of Physiopathology Molecular Genetic and Biotechnology. Ain Chock Faculty of Sciences, Hassan II University, Casablanca. And Anfa Fertility Center Private Clinic of in vitro fertilization and Endoscopic Surgery, Casablanca. Methodology: We included 101 men patients subdivided into three groups. Group A with normal sperm parameters and a positive pregnancy; Group B with normal sperm parameters and a negative pregnancy; Group C with abnormal sperm parameters and a negative pregnancy. After DGC, lipid peroxidation (MDA) level, as well as the antioxidant enzyme activities of superoxide dismutase (SOD), glutathione reductase (GR) and catalase (CAT), were evaluated. Results: Patients with normal and abnormal sperm parameters showed that MDA level, and antioxidant enzymes activities increased significantly from the pellet, to 80 %, and from 80% to 40% fractions. In addition, lipid peroxidation and enzymes activities levels were significantly lower in patients with positive pregnancy than in patients with negative pregnancy and especially those with abnormal spermatic parameters. Conclusion: The DGC would select sperm not only motile and viable but with less oxidative stress. Moreover, the rate of lipid peroxidation and antioxidant enzyme activities could differentiate between patients with normal and abnormal spermatic parameters and between patients with positive and negative pregnancy outcome.

Scorpion envenomation is a serious public health issue.

Background The aim of this study is to assess the corrosion resistance behaviour of Nickel-Titanium-based orthodontic wires (NiTi) in different concentrations of Sodium Fluoride (NaF) and the corrosion's inhibitory effect of the extracts of some medicinal plants (essential oils, hydrosols and extract). Material and methods In this study we used NiTi (3M) and CuNiTi (ORMCO, 35°C, California) orthodontic wires. The following electrolytes were prepared: Lactate Ringer solution with additions of 0.1%, 0.5% or 1% of Sodium Fluoride and the extracts of different plants: Artemisia, Syzygium aromaticum (Clove) and Celtis australis. Corrosion resistance was studied using anodic potentiodynamic polarisation and electrochemical impedance spectroscopy measurements. At the end of the experiment, microscopic images of wires were performed. ANOVA test with the comparison of Bonferroni and Tukey tests were performed to elucidate comparisons among all groups. Results The higher sodium fluoride concentration is related to negative corrosion potential for both NiTi and CuNiTi orthodontic wire. Hydrosols are associated to positive values of corrosion potential. CuNiTi has a lower corrosion resistance than NiTi. Conclusions The prescription of toothpastes containing sodium fluoride should be reduced especially for patients wearing fixed orthodontic appliances. Eugenol may be considered as alternative of sodium fluoride for orthodontic patients for its anti-microbial and anti-corrosive effects. Key words:Corrosion behaviour, Sodium Fluoride, Nickel-Tatanium, orthodontic wires, corrosion inhibitors, aromatical plants.

Hepatocellular carcinoma (HCC) is the most common primary liver cancer in adults, the fifth most common malignancy worldwide and the third leading cause of cancer related death. An alternative to the surgical treatments and drugs, such as sorafenib, commonly used in medicine is necessary to overcome this public health problem. In this study, we determine the anticancer effect on HCC of Moroccan cobra Naja haje venom and its fraction obtained by gel filtration chromatography against Huh7.5 cancer cell line. Cells were grown together with WI38 human fibroblast cells, LX2 human hepatic stellate cell line, and human endothelial cells (HUVEC) in MCTS (multi-cellular tumor spheroids) models. The hepatotoxicity of venom and its fractions were also evaluated using the normal hepatocytes cell line (Fa2N-4 cells). Our results showed that an anti HCC activity of Moroccan cobra Naja haje venom and, more specifically, the F7 fraction of gel filtration chromatography exhibited the greatest anti-hepatocellular carcinoma effect by decreasing the size of MCTS. This effect is associated with a low toxicity against normal hepatocytes. These results strongly suggest that the F7 fraction of Moroccan cobra Naja haje venom obtained by gel filtration chromatography possesses the ability to inhibit cancer cells proliferation. More research is needed to identify the specific molecule(s) responsible for the anticancer effect and investigate their mechanism of action.

International audience; Background: Control of the mosquito vector Culex pipiens with insecticides is the main way to control arboviruses that the species can transmit such as West Nile virus (WNV) and Rift Valley fever virus (RVFV). However, its efficiency has been hampered by the emergence of insecticide resistance. Little is known about the insecticide-resistance status and underlying resistance mechanisms of field-collected populations of Cx. pipiens in Morocco.Methods : Mosquito adults from Mohammadia city in Morocco were reared from immature stages. The level of their susceptibility to insecticides was assessed using standard WHO bioassay. The two forms of the Cx. pipiens complex and their hybrids were identified by a multiplex PCR. Identified mosquitoes were then tested for the presence of the G119S ace-1 and L1014F kdr mutations using PCR-RFLP and PCR assays, respectively.Results : WHO bioassays indicated that Cx. pipiens was resistant to all tested insecticides: lambda-cyhalothrin (49% mortality), permethrin (63% mortality), DDT (16% mortality), malation (52% mortality) and bendiocarb (39% mortality). The frequency of the 119S allele was almost identical in the pipiens form and hybrids (0.11 and 0.15, respectively) whereas it remained low in the molestus form (0.03). No significant correlation was observed between the G119S allele and the resistance phenotype to two tested insecticides (malathion and bendiocarb). The frequency of the L1014F allele was identical in the pipiens form and hybrids (0.44) whereas it was low in the molestus form (0.36) but no significant difference was detected (χ2 = 1.46, df = 1, P = 0.225). The presence of the L1014F kdr mutation was significantly associated with resistance to three tested insecticides in pipiens form (P = 0.0019, P = 0.0023 and P = 0.023, respectively, to lambda-cyhalothrin, permethrin and DDT) whereas no significant correlation was observed between the L1014F kdr mutation and resistance phenotype in molestus form and hybrids to the three tested insecticides.Conclusion : These findings showed that wild populations of Cx. pipiens have developed resistance against the main insecticide families with different modes of action: organochlorines (DDT), organophosphates (malathion), carbamates (bendiocarb), pyrethroids (lambda-cyhalothrin, permethrin). Therefore, urgent action should be taken to manage the resistance in this species to maintain the effectiveness of arbovirus control.

Exposure to paraphenylenediamine (PPD), a derivative of paranitroaniline widely used as an oxidisable hair dye, is often associated with the development of allergic contact dermatitis. Such reactions involve activation of the subject’s immune system and it is known that neutrophils are the first cells to arrive at the site of perturbation. In view of its pivotal role in the initiation and amplification of inflammation, the fate of the neutrophil in presence of PPD requires attention. Thus, the purpose of the present study was to evaluate the vitro effects of PPD on human neutrophils viability and on reactive oxygen species production. Neutrophils from healthy volunteers were incubated with three concentrations of PPD (11.5, 23 and 46 µM). Apoptosis was evaluated by light microscopy and DNA gel electrophoresis, and oxidative stress was evaluated by spectrophotometry. Results showed that PPD induces neutrophils apoptosis in a dose and time dependent manner with a LC50 value of 23 µM at 6 h of treatment. Furthermore, at 5 h of treatment, PPD markedly increased lipid peroxidation (47%) and enhanced the activity of catalase, glutathione reductase and superoxide dismutase. These results suggest that PPD can induce neutrophils apoptosis and an oxidative stress. Key words: Paraphenylenediamine, neutrophils, apoptosis, oxidative stress, catalase, lipid peroxidation, glutathione reductase, superoxide dismutase.

Since reactive oxygen species (ROS) play an important role in the pathogenesis of many human diseases, attention is given to the development of safe and effective natural antioxidants. In the present study, the protective and antioxidant potential of the argan oil extracted from the fruit of Argania spinosa, an endemic tree in Morocco, was evaluated against hydrogen peroxide (H2O2) induced oxidative damages in Tetrahymena pyriformis. The protozoan cells were cultivated in polypeptone yeast glucose (PPYG) medium supplemented with argan oil (AO) at a dose of 0.5% (v/v) during 4 subcultures and treated with different concentrations of H2O2. The effect of this oil on the protozoan growth was followed and the antioxidant enzyme activities of catalase (CAT), superoxyde dismutase (SOD) and glutathione reductase (GR) were evaluated as well as lipid peroxydation (MDA). As compared to the controls, the AO increased both the cell viability and the antioxidant enzyme activities and decreased the MDA level. The co incubation of T. pyriformis with AO and H2O2 (300 µM) reduced the H2O2 cytotoxicity. The cells cultivated in the presence of AO then exposed to H2O2 for 3 h, show a significant reduction in MDA level (p < 0.001), an important increase in CAT (p

International audience; 2,4-Dichlorophenoxyacetic acid (2,4D) is one of the widely used herbicide of the phenoxy family with possible startling number of adverse effects on species other than the weeds which is designed to kill. The effects of 2,4D were investigated in jerboa (Jaculus orientalis), a wild animal of subdesert highlands. The jerboas have been daily treated intraperitonally with 2,4D 3 mg/kg body weight for 4 weeks. Plasmatic markers, and antioxidants defences systems were assessed and histological alterations were evaluated. The in vivo and in vitro effects of 2,4D on the mitochondrial D-3-hydroxybutyrate dehydrogenase (BDH) were also determined. Our results showed a strong decrease of triglycerides level and HDL cholesterol and an increase in GOT level and LDL cholesterol. The microscopic evaluation showed that 2,4D induced necrosis of seminiferous tubules cells in testis, hyperplasia of hepatocytes in liver and presence of multinucleated giant cells in brain. The results show also an inhibitory effect on BDH in terms of activity and kinetic parameters. All of these results show that 2,4D induces toxicity which affects energy metabolism, morphological perturbation and oxidative stress.

International audience; Ciprofibrate is a well-known drug used to normalize lipid parameters and fibrinogen in atherosclerosis patients. In laboratory rodents such as rats or mice, ciprofibrate exhibits peroxisome proliferator activity. However, to date, no clear alterations or side effects caused by ciprofibrate have been noted in humans. In order to further investigate such possible relationships, we studied the effects of sustained ciprofibrate treatment in jerboas (Jaculus orientalis). In these rodents, ciprofibrate does not induce hepatomegaly or promote liver cell DNA replication, confirming that this species more closely resembles humans than do rats or mice. The jerboas were treated daily with ciprofibrate at 3 mg/kg body weight for 4 weeks. Subcellular markers, clinical enzymes and enzymatic antioxidant defenses were then assessed. The results showed a strong decrease in peroxisomal catalase activity and an increase in the level of malondialdehyde (a stress biomarker). Moreover, ciprofibrate in vivo and in vitro inhibited D-3-hydroxybutyrate dehydrogenase, a mitochondrial enzyme of the ketone body interconversion that is important in redox balance (NAD+/NADH+H+ ratio). In conclusion, under these conditions, ciprofibrate induced alterations in the liver oxidative metabolism.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) was purified from two amphibian species, Xenopus laevis and Pleurodeles waltl. Comparative studies revealed that the two proteins differ by their subunit molecular masses, pI values and V8 digested peptide maps. The effect of zinc, cadmium and copper ions on GAPDH enzymatic activity has been examined in vitro. A time, metal concentration and metal type dependent inhibition was observed for both enzymes. X. laevis and P. waltl GAPDHs exhibit a much greater sensitivity to copper than to cadmium or zinc ions. Different half-lives and differential sensitivity to various metals was observed between the two enzymes with P. waltl GAPDH being remarkably tolerant to cadmium ions compared to the X. laevis enzyme. In order to understand the differential sensitivity of the two enzymes to metals, we produced 3D models of both X. laevis and P. waltl GAPDH structures based upon known 3D structures of GAPDHs from other species. This necessitated, in a first step, to clone a 900 bp cDNA fragment encoding the nearly full-length P. waltl GAPDH. Spatial motif searches on the homology models indicated potential metal binding sites involving cysteine and histidine residues outside the catalytic sites, existing only in either the X. laevis or the P. waltl GAPDH sequences.

The characterization of metallothionein in the Urodele amphibian species Pleurodeles waltl was achieved. A simple and rapid method for identification of metallothionein, based on its strong affinity for cadmium (109Cd), was used. We were able to show that metallothionein is constitutively synthesized in liver, ovary and brain. The property of metallothionein to strongly bind essential (Zn, Cu) as well as toxic (Cd, Hg) metals is consistent with a dual role in cellular metabolism, i.e. homeostatis and detoxification of heavy metal ions.

The NAD(+)-dependent cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) has been purified to homogeneity from skeletal muscle of the newt Pleurodeles waltl (Amphibia, Urodela). The purification procedure including ammonium sulfate fractionation followed by Blue Sepharose CL-6B chromatography resulted in a 24-fold increase in specific activity and a final yield of approximately 46%. The native protein exhibited an apparent molecular weight of approximately 146 kDa with absolute specificity for NAD(+). Only one GAPDH isoform (pI 7.57) was obtained by chromatofocusing. The enzyme is an homotetrameric protein composed of identical subunits with an apparent molecular weight of approximately 37 kDa. Monospecific polyclonal antibodies raised in rabbits against the purified newt GAPDH immunostained a single 37-kDa GAPDH band in extracts from different tissues blotted onto nitrocellulose. A 510-bp cDNA fragment that corresponds to an internal region of a GapC gene was obtained by RT-PCR amplification using degenerate primers. The deduced amino acid sequence has been used to establish the phylogenetic relationships of the Pleurodeles enzyme--the first GAPDH from an amphibian of the Caudata group studied so far--with other GAPDHs of major vertebrate phyla.

Mice were immunized with membrane preparations of epidermal cells taken from different parts (internal and external face of femur and apex and base of tibia) of the metathoracic legs of cockroach larvae. Using indirect immunofluorescence, anti-internal face of femur antibodies were observed to bind preferentially to membranes from the internal face of the femur; similarly, anti-external face of femur antibodies bound preferentially to membranes from the external face of the femur. We also found a preferential binding of anti-apex of tibia antibodies to membranes from the apex of the tibia and anti-base of the tibia antibodies to membranes from the base of the tibia. When anti-tibia sera were tested on membranes from the femur, anti-apex of tibia antibodies bound preferentially to membranes from the apex of the femur, and anti-base of tibia antibodies bound preferentially to membranes from the base of the femur.This demonstrates that epidermal cell membranes from the different parts of the leg differ in their antigenic properties, and that these differences are related to their position around the appendage and along the proximodistal axis of segments.These results are in agreement with those of previous graft experiments and with the concept of ordered sequences in insect appendages.