Search

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.

Buthus occitanus (B. occitanus) is one of the most dangerous scorpions in the world. Despite the involvement of B. occitanus scorpion in severe cases of envenomation in Morocco, no study has focused yet on the proteomic composition of the Moroccan B. occitanus scorpion venom. Mass spectrometry‐based proteomic techniques are commonly used in the study of scorpion venoms. The implementation of top‐down and bottom‐up approaches for proteomic analyses facilitates screening by allowing a global view of the structural aspects of such complex matrices. Here, we provide a partial overview of the venom of B. occitanus scorpion, in order to explore the diversity of its toxins and hereafter understand their effects. To this end, a combination of top‐down and bottom‐up approaches was applied using nano‐high liquid chromatography coupled to nano‐electrospray tandem mass spectrometry (nano‐LC‐ESI MS/MS). The LC‐MS results showed that B. occitanus venom contains around 200 molecular masses ranging from 1868 to 16 720 Da, the most representative of which are those between 5000 and 8000 Da. Interestingly, combined top‐down and bottom‐up LC‐MS/MS results allowed the identification of several toxins, which were mainly those acting on ion channels, including those targeting sodium (NaScTxs), potassium (KScTxs), chloride (ClScTxs), and calcium channels (CaScTx), as well as antimicrobial peptides (AMPs), amphipathic peptides, myotropic neuropeptides, and hypothetical secreted proteins. This study reveals the molecular diversity of B. occitanus scorpion venom and identifies components that may have useful pharmacological activities., Buthus occitanus scorpion possesses highly toxic venom. This article describes the first proteomic screening of the Moroccan B. occitanus venom. Our findings highlight the complexity and the toxin diversity of this venom. The most representative compounds are neurotoxins. In addition, antimicrobial peptides, amphipathic peptides, myotropic neuropeptides, and hypothetical secreted proteins were also detected in this venom.

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.

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

Alcohol intake and cigarette smoking are the major lifestyle factors with negative impact on fertility. We were interested to evaluate the negative impact of these factors on oxidative stress (OS), enzymatic antioxidant activity (EAO) of spermatozoa and on its DNA damage. This study included 108 male infertile patients with normal range of sperm conventional parameters but with unexplained infertility in assisted reproductive technologies programme. Firstly, OS was analysed based on lipid peroxidation (MDA) and EAO which included catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR). Secondly, we evaluated DNA fragmentation by TUNEL assay and chromatin decondensation by aniline blue colouration. The whole lot was divided into four groups: control (nonalcoholic and nonsmoker patients), alcohol group, smoking group and alcohol-smoking group. The results showed, in three last groups compared to control an increased CAT, SOD and GR activities with high MDA level especially in smoking and alcohol-smoking group. The latter showed the highest values of DNA fragmentation and chromatin decondensation (31% and 39%) to exceed DNA damage normal range. Indeed, smoking and alcohol intake lead to increase EAO due to long-term unbalanced antioxidant/oxidation ratio with high OS which cause consequently sperm DNA damage calling in need by urgency to change the lifestyle behaviour.

Paraphenylenediamine (p-PD) is the main aromatic amine used in the formulation of hair dyes. Epidemiologic studies have suggested that the use of p-PD based hair dyes might be related to increase risk of human malignant tumors including multiple myeloma and hematopoietic cancers. However, the toxicity of p-PD on myeloma cells has not been well elucidated yet. Therefore, the association between the cytotoxicity of p-PD and the reactive oxygen species (ROS) generation on murine myeloma cells P3X63Ag8.653 (P3) was evaluated. Treatment with p-PD decreases cell viability in a dose and time dependent manner. In addition, p-PD markedly enhanced lipid peroxidation. This increase was accompanied with a decrease in both glutathione reductase and superoxide dismutase activities. Furthermore, Pre-treatment of P3 cells with antioxidants, reduced glutathione or manganese II chloride, significantly inhibited p-PD induced cytotoxicity and ROS generation. Based on these results, p-PD might induce apoptosis via the involvement of ROS. Key words: Murine myeloma cells, p-Phenylenediamine, apoptosis, oxidative stress, ROS, antioxidants.

The present study was undertaken to validate the benefits of a fluoropolymer treatment on the biostability, inflammatory response, and healing characteristics of a polyester mesh used for hernia repair, the Fluoromesh, as compared to a commercial monofilament-knit polypropylene mesh, Marlex, used as the control. Both were implanted for the repair of surgically induced abdominal hernias in piglets for prescheduled durations of implantation of 4, 15, and 60 days. The mesh and surrounding tissue were harvested at the sacrifice for the bursting strength and inflammatory response measurements in terms of alkaline and acid phosphatase secretion in the tissue, and for histological observations of the healing sequence and tissue thickness measurements by histomorphometric techniques. After cleaning to remove adherent tissue, the presence of the fluoropolymer at the surface of the mesh was detected using SEM and ESCA. The results demonstrated greater mechanical reinforcement and tissue development for the Fluoromesh than for the polypropylene mesh. The healing performance of the Fluoromesh was attributed to a more intense chronic inflammatory reaction early after implantation that stimulated significantly greater tissue ingrowth and integration. The concentration of fluoropolymer at the surface of the mesh was masked as a result of biological species adsorption. Textile analysis revealed that the Fluoromesh was dimensionally more stable in vivo than the polypropylene control mesh, which demonstrated stretching in the weft direction and shrinking in the warp direction during implantation.

We have evaluated the efficacy of 13 analogues of retinoic acid (retinoids) in producing in vivo feathers on the normally scaly foot integument (ptilopody) of the chick embryo. The retinoids were dissolved in dimethylsulfoxide and injected at doses from 1.25-250 micrograms into the amniotic cavity of embryos of 10 days of incubation. The results showed that the production of ptilopody requires specific retinoids, namely, those incorporating a free trans carboxylic end group. Indeed, the esterification of this acid end group (retinoid ethyl ester) leads to a delay in the effect of the product. Its substitution by an alcohol (retinol) or an aldehyde group (retinal), or its isomerization, leads to inactive products. The esterification by acetic acid of the alcohol terminal group of retinol leads to a moderately active compound (retinyl acetate). By contrast, modifications of the ring do not interfere with the efficacy of the retinoid, and modifications of the side chain by addition of one or two supplementary rings lead to an increase of the activity.

Feet of chicks are normally covered with scales. Injection of retinoic acid into the amniotic cavity of 10-day chick embryos causes the formation of feathers on the foot scales. To elucidate whether retinoic acid affects primarily the epidermis or the dermis, heterotypic dermal-epidermal recombinants of tarsometatarsal skin were tested as to their morphogenetic capacity, when grafted to the chick chorioallantoic membrane. Recombinants involving treated epidermis and untreated dermis formed feathered scales, while the reverse recombinants of untreated epidermis and treated dermis led to the formation of scales only. Likewise the association of treated tarsometatarsal dermis with untreated epidermis from a non-appendage-forming region (the midventral apterium) resulted in the formation of scales only. These results show that retinoic acid affects primarily the epidermis. Further insight into the mechanism of dermal-epidermal interaction was gained by heterotopic recombinations of early (8.5- and 10-day) untreated tarsometatarsal dermis with epidermis from the midventral apterium. These recombinants formed scales, proving that tarsometatarsal dermis is endowed with scale-forming properties as early as 8.5 days of incubation. Finally, it is concluded that retinoic acid acts on the chick foot epidermal cells by temporarily inhibiting their scale placode-forming properties, allowing their latent feather placode-forming properties to be expressed.