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SummaryCell-free DNA (cf-DNA) is defined as DNA fragments that are released into the body fluids from apoptosis or necrosis cells, including follicular fluid (FF), which can affect the microenvironment of the oocyte associated with infertility. We aimed to investigate a relationship between apoptosis of cumulus cells (CCs) and cf-DNA levels in FF and clinical outcomes of women undergoing intracytoplasmic sperm injection (ICSI). Therefore, 82 FF samples were collected, and the corresponding CCs were isolated for ICSI procedures. FF cf-DNA concentration was quantified using ALU-quantitative polymerase chain reaction (PCR), and CCs DNA fragmentation index (DFI) was evaluated by the terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labelling (TUNEL) method. We found that cf-DNA and DFI levels were significantly higher in FF and CCs samples related to the age of women ≥37 years compared with the age of women < 37 years. Moreover, in older and younger women, FF cf-DNA and CCs DFI levels were significantly lower when the anti-Müllerian hormone (AMH) level was > 1.1 ng/ml compared with when AMH ≤ 1.1 ng/ml. In addition, patients with a low number of retrieved oocytes ≤ 6 had significantly higher levels of CCs DFI and FF cf-DNA than women with a higher number of retrieved oocytes > 6. Additionally, we observed that higher levels of cf-DNA and DFI were associated with poor oocyte maturity and poor embryo quality. Finally, cf-DNA and DFI levels were significantly lower in pregnant women than in non-pregnant ones. We conclude that DFI and cf-DNA levels in the oocyte microenvironment could have potential use in evaluating oocyte and embryo developmental competence.

Background: The success of in vitro-fertilization (IVF) cycles is determined in large part by the quality of embryo cleavage, which in turn, is dependent on the quality of the embryo culture media (CM). Many factors can influence the quality of embryo CM, one of which is the levels of Cell Free Deoxyribonucleic acid (DNA). Understanding the association between Cell-free DNA levels in embryo CM and the quality of embryo cleavage could help improve the quality of IVF techniques. Methods: This prospective study was conducted with 96 spent CM from patients undergoing IVF cycle, in order to determine relationships of Cell-free DNA levels in embryo CM with embryo cleavage quality on day 3. After intracytoplasmic sperm injection (ICSI), 48 embryos were evaluated on day 3 of their development, according to their cell number. Day 2 and day 3 CM corresponding to each one of the embryos was analyzed, by quantitative PCR, for estimation of Cell-free DNA levels. Results: The results revealed a significant increase in Cell-free DNA levels on day 2 CM corresponding to 4 to 6 cell embryos compared to those corresponding to 7 to 8 cell embryos (p=0.04). As for day 3 CM, the results showed no significant difference between the Cell-Free DNA levels in CM of 7-8 and those of 4-6 cell embryos (p=0.4). Also, cell free DNA levels in embryo CM, were significantly higher on day 2 compared to day 3 for both 7-8 and 4-6 cell embryos (p=0.03; p=0.04). Conclusion: We conclude that cell-free DNA levels in CM might be associated with delayed embryo cleavage.

Background: The process of preimplantation embryo development in vitro represents a key phase during in vitro fertilization (IVF) cycles. It involves several regulatory signaling pathways as well as an optimized in vitro culture system. The resulting embryo quality helps to determine embryo competence before implantation and pregnancy outcomes. Reactive oxygen species (ROS) are known to play a major role in influencing the process of embryo development. Their role can be reflected in the regulation of signaling pathways as second messengers or in the irreversible cell alterations due to oxidative stress following an excess of ROS levels. Methods: In this study, we investigated the association between morphological embryo quality (fertilization, cleavage quality, and fragmentation levels) and lipid peroxidation levels (Malondialdehyde) in embryo culture media. After intracytoplasmic sperm injection (ICSI), a total of 103 oocytes were evaluated on day 1 and day 3 of their development, and their corresponding culture media were analyzed by estimating MDA levels using thiobarbituric acid. Results: The results showed no significant association between MDA levels in culture media and fertilization rate (p=0.3), sperm quality (p=0.99; p= 0.17; p=0.46; p=0.30; p=0.65; p=0.44; p=0.09; p=0.15; p=0.56), embryo fragmentation levels (p=0.79; p=0.40), AMH levels (p=0.31; p=0.36) and female age (p=0.60; p=0.34). However, we revealed a significant association between cleavage quality and MDA levels in the embryo environment (p=0.03). Conclusion: We conclude that oxidative stress in IVF culture media might be mainly associated with delayed embryonic development.

The present study aimed to assess the prognostic interest of metabolic and anatomic parameters derived from 2-deoxy-2-[In this study, pre-treatment [Our results revealed a significant association between pre-treatment nodal-maximum standardized uptake value (N-SUV max) and N categories (p = 0.01), between pre-treatment node-to-tumor SUV ratio (NTR) and both tumor size (p = 0.01) and N categories (p = 0.009), as well as between metabolic tumor volume (MTV) and both tumor size and NPC overall stage (p 0.000). In multivariate analyses, pre-treatment N-SUV max, NTR and MTV were significant independent predictors of overall survival, distant metastasis-free survival, and progression-free survival (PFS) (p 0.05). N-SUV max and MTV were also found to be significant independent predictors of loco-regional recurrence-free survival (p 0.05), whereas HN-MRI detection of skull-base bone invasion was an independent factor associated with worse PFS in NPC (p = 0.03).The present study highlights N-SUV max, NTR and MTV derived from [

Background: Androctonus mauretanicus (Am ) is one of the most hazardous scorpions in Morocco and has a highly toxic venom responsible for severe cases of envenomation. However, few studies have focused on deciphering its proteic composition. Objectives: Herein, we aim to map out the complete proteome of the Am venom filtrate to highlight its complexity and the polymorphism of its toxic content. This, in turn, will lead to a deeper understanding of the toxins’ mechanism of action and will help uncover those with therapeutic potential. Methods: Top-down and bottom-up proteomic approaches were used complementarily to decipher the proteome of the Am venom. These approaches were carried out on nano-high liquid chromatography coupled to nano-electrospray tandem mass spectrometry (Nano-LC-ESI-MS/MS). Results: Am venom encloses a complex mixture of 269 different compounds with molecular weights ranging from 1618.74 to 14 214.84 Da. The most abundant ones showed masses from 6185.92 to 7899.53 Da (53.89%) followed by those ranging from 2079.25 to 5969.63 Da (37.81%). Interestingly, the combination of the results of both approaches allowed the screening of a total of 112 peptides. The highest percentage was represented by neuropeptides (87%), including NaTxs, KTxs, ClTxs, venom proteins, venom neuropeptides, and myotropic neuropeptides. Moreover, other peptides were identified, such as antimicrobial peptides, amphipathic peptides, cysteine-rich venom peptides, enzymes, kunitz-type inhibitors and orphan peptides. Conclusion: The Am venom appears to contain a great number of diverse peptides, some of which could prospectively be exploited for their pharmaceutical potential.

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.

SummaryThe process of embryonic development is crucial and radically influences preimplantation embryo competence. It involves oocyte maturation, fertilization, cell division and blastulation and is characterized by different key phases that have major influences on embryo quality. Each stage of the process of preimplantation embryonic development is led by important signalling pathways that include very many regulatory molecules, such as primary and secondary messengers. Many studies, bothin vivoandin vitro, have shown the importance of the contribution of reactive oxygen species (ROS) as important second messengers in embryo development. ROS may originate from embryo metabolism and/or oocyte/embryo surroundings, and their effect on embryonic development is highly variable, depending on the needs of the embryo at each stage of development and on their environment (in vivoor underin vitroculture conditions). Other studies have also shown the deleterious effects of ROS in embryo development, when cellular tissue production overwhelms antioxidant production, leading to oxidative stress. This stress is known to be the cause of many cellular alterations, such as protein, lipid, and DNA damage. Considering that the same ROS level can have a deleterious effect on the fertilizing oocyte or embryo at certain stages, and a positive effect at another stage of the development process, further studies need to be carried out to determine the rate of ROS that benefits the embryo and from what rate it starts to be harmful, this measured at each key phase of embryonic development.

Les maladies cardiovasculaires (MCV) sont la première cause de mortalité dans le monde. Les plaquettes jouent un rôle majeur dans le développement de ces maladies et la mise au point d’antiplaquettaires efficaces constitue une priorité dans le cadre de la lutte contre la mortalité liée aux MCV. L’aspirine et les médicaments de la famille des thiénopyridines sont les agents antiplaquettaires les plus utilisés actuellement. Ces médicaments ciblent des voies de signalisation impliquées dans l’initiation de l’agrégation, exerçant ainsi un effet antiplaquettaire modéré. D’autres médicaments aux effets plus importants, comme les molécules dirigées contre le récepteur GPIIb/IIIa, inhibent l’agrégation plaquettaire indépendamment de la voie de signalisation initiant l’activation plaquettaire, mais ils sont associés à des complications hémorragiques majorées. Étant données les caractéristiques spécifiques de chacun de ces agents antiplaquettaires, leur prescription nécessite de prendre en compte le type d’évènement cardio-vasculaire, l’âge et les comorbidités du patient traité et, bien sûr, les effets secondaires hémorragiques potentiels de la molécule qui est prescrite. Apparaît donc la nécessité de mettre au point de nouvelles molécules ayant un effet plus ciblé, gardant une efficacité optimale, mais permettant une réduction du risque hémorragique qui constitue la principale limite des médicaments antiplaquettaires.

Scorpion envenomation is a serious public health issue.

The present meta-analysis was conducted to evaluate the prognostic value of pre and post-Epstein Barr Virus (EBV) DNA load testing and to assess the clinical benefit of using this molecular approach in the prognosis for a better nasopharyngeal carcinoma (NPC) management. Relevant studies were searched in different database until May 2020. Patient´s outcomes overall survival (OS), disease free survival (DFS), progression-free survival (PFS), distant-metastasis-free survival (DMFS), and local-regional-failure-free survival (LRFS), hazard ratios (HRs) and 95% confidence intervals (CIs) were extracted from selected studies. The association of pre and post-EBV DNA load and survival outcomes was assessed using review manager and the pooled HRs with 95% CIs were calculated. Twenty-six eligible studies were included in this meta-analysis, with a total of 9966 patients. Pooled HRs showed that EBV DNA levels before and after treatment are significantly associated with survival outcomes, with HR (95% CI) of 2.09 [1.74, 2.51] for OS, 1.77 [1.19, 2.62] for DFS, 2.53 [2.18, 2.92] for DMFS, 1.78 [1.45, 2.19] for LRFS and 2.17 [1.91, 2.47] for PFS in pre-EBV DNA, and an HR (95%) of 4.52 [2.44, 8.36], 4.08 [2.38, 6.99], 5.59 [ 3.58, 8.71] and 8.88 [5.29, 14.90] for OS, DFS and PFS and DMFS in post-EBV DNA, respectively. High pre and post-EBV DNA levels were significantly associated with poor NPC patient´s survival outcomes; which clearly confirm the high interest to introduce viral EBV DNA load as a prognostic biomarker for NPC management.

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.

Purpose: While nasopharyngeal carcinoma (NPC) management in Morocco is still based on conventional work-ups: a head and neck computed tomography (HN-CT), thoracic and abdominal CT and bone scan, the combination of HN magnetic resonance imaging (HN-MRI) and 2-Deoxy-2-[18F] fluoro-D-glucose positron emission tomography/computed tomography ([18F] FDG PET/CT) is now widely used in the diagnostic and follow-up of this malignancy.Methods: In this prospective study, [18F] FDG PET/CT and HN-MRI outcomes of 117 NPC patients diagnosed between January 2017 and December 2018 were investigated in order to assess their usefulness in routine management of Moroccan patients with NPC. The concordance between HN-MRI and [18F] FDG PET/CT in Tumor (T) and Nodal (N) classification was assessed and the association between [18F] FDG PET/CT metabolic parameters (Tumor- maximum standardized uptake value (T-SUV max), Nodal (N-SUV max), node-to-tumor SUV ratio (NTR) and distant metastasis (M-SUV max), TNM staging system, NPC stages and patient’s survival outcomes was evaluated. Results: Our results showed a moderate concordance between T-TEP and T-MRI categories with a Cohen kappa coefficient (k) at 0.45, and a mediocre concordance between N-TEP and N-MRI (k=0.3). Metabolic parameters of the [18F] FDG PET/CT were assessed; N-SUV max values were significantly higher in patients with advanced nodal involvement, with a mean of 7.4, 9.7 and 11.0 for patients with N1, N2 and N3 nodal categories, respectively (pConclusion: Our findings provide additional evidence into the complementary roles of HN-MRI and [18F] FDG PET/CT in TNM and overall staging of NPC. To the best of our knowledge, this is the first Moroccan study to highlight N-SUV max and NTR derived from [18F] FDG-PET/CT as promising metabolic biomarkers for NPC prognosis.

Background The identification of effective prognosis biomarkers for nasopharyngeal carcinoma (NPC) is crucial to improve treatment and patient outcomes. In the present study, we have attempted to evaluate the correlation between pre-treatment plasmatic Epstein-Barr virus (EBV) DNA load and the conventional prognostic factors in Moroccan patients with NPC. Methods The present study was conducted on 121 histologically confirmed NPC patients, recruited from January 2017 to December 2018. Circulating levels of EBV DNA were measured before therapy initiation using real-time quantitative PCR. Results Overall, undifferentiated non-keratinizingcarcinoma type was the most common histological type (90.1 %), and 61.8 % of patients were diagnosed at an advanced disease stage (IV). Results of pre-treatment plasma EBV load showed that 90.9 % of patients had detectable EBV DNA, with a median plasmatic viral load of 7710 IU/ml. The correlation between pre-treatment EBV DNA load and the conventional prognostic factors showed a significant association with patients’ age (p = 0.01), tumor classification (p = 0.01), lymph node status (p = 0.003), metastasis status (p = 0.00) and overall cancer stage (p = 0.01). Unexpectedly, a significant higher level of pre-treatment EBV DNA was also found in plasma of NPC patients with a family history of cancer (p = 0.04). The risk of NPC mortality in patients with high pretreatment EBVDNA levels was significantly higher than that of those with low pre-treatment plasma EBV-DNA levels (p p Conclusions The results of the present study clearly showed a high association between pre-treatment EBV DNA load, the crucial classical prognostic factors (T, N, M and disease stage) of NPC and OS, suggesting that pre-treatment EBV DNA can be a useful prognostic biomarker in clinical decision-making and improving NPC treatment in Morocco.

Cardiovascular diseases are the leading cause of deaths in the world. Platelets play a major role in the occurrence of these diseases and the development of antiplatelet drugs is a priority in the fight against cardiovascular diseases-associated mortality. Aspirin and thienopyridine-based P2Y12 inhibitors are the main drugs currently used. These molecules target the initiation of platelets activation and are responsible for a moderate inhibitory action. Other antiplatelet agents, as glycoprotein (GP) IIb/IIIa antagonists, inhibit platelet aggregation independently of initial activation-associated pathways, but are responsible for increased hemorrhagic events. Regarding each antiplatelet agent's specific characteristics, the prescription of these drugs must take into account the type of cardiovascular event, the age of the patient, the past medical history, and the potential hemorrhagic adverse events. Thus, there is a need for the development of new molecules with a more targeted effect, maintaining optimal efficiency but with a reduction of the hemorrhagic risk, which is the principal limitation of these treatments.Antiplaquettaires actuels, en cours de développement et cibles thérapeutiques.Les maladies cardiovasculaires (MCV) sont la première cause de mortalité dans le monde. Les plaquettes jouent un rôle majeur dans le développement de ces maladies et la mise au point d’antiplaquettaires efficaces constitue une priorité dans le cadre de la lutte contre la mortalité liée aux MCV. L’aspirine et les médicaments de la famille des thiénopyridines sont les agents antiplaquettaires les plus utilisés actuellement. Ces médicaments ciblent des voies de signalisation impliquées dans l’initiation de l’agrégation, exerçant ainsi un effet antiplaquettaire modéré. D’autres médicaments aux effets plus importants, comme les molécules dirigées contre le récepteur GPIIb/IIIa, inhibent l’agrégation plaquettaire indépendamment de la voie de signalisation initiant l’activation plaquettaire, mais ils sont associés à des complications hémorragiques majorées. Étant données les caractéristiques spécifiques de chacun de ces agents antiplaquettaires, leur prescription nécessite de prendre en compte le type d’évènement cardio-vasculaire, l’âge et les comorbidités du patient traité et, bien sûr, les effets secondaires hémorragiques potentiels de la molécule qui est prescrite. Apparaît donc la nécessité de mettre au point de nouvelles molécules ayant un effet plus ciblé, gardant une efficacité optimale, mais permettant une réduction du risque hémorragique qui constitue la principale limite des médicaments antiplaquettaires.

Background: Intellectual disability (ID) has been defined as a considerably reduced ability to understand new or complex information and to learn new skills. It is associated with life-long intellectual and adaptive functioning impairments that have a profound impact on individuals, families, and society. It affects about 3% of the general population. ID often comes out with other mental conditions like attention deficit, hyperactivity, and autism spectrum disorders (ASD), and it can be part of a malformation syndrome that affects other organs. It may be syndromic (S-ID) or non-syndromic (NS-ID). Objective: The aims of this study were to identify the profile of intellectually disable patients being referred for cytogenetic analysis in Morocco, to determine the prevalence of chromosomal abnormalities in a Moroccan group, and to compare the results with those of analogous studies from other countries. Participants: We included data from Moroccan patients with NS-ID and others with S-ID (mostly Down syndrome cases) who have been referred between 1996 and 2016. 1,626 patients were involved in this study, 1,200 were referred with a clinical diagnosis of Down syndrome, 37 were clinically diagnosed for ASD with ID, and 389 were suspected of NS-ID. Results: We identified 1,200 cases of Down syndrome. In 1,096 analyses (91.3%), a cytogenetic variant of trisomy 21 was identified: standard trisomy 21 in 1,037 cases (94.6%), a translocation in 34 cases (3.10%), and mosaicism in 25 cases (2.3%). The cytogenetic analysis among ASD with ID cases did not reveal any specific chromosomal abnormalities. The present study also shows that chromosomal abnormalities were present in 6.43% of the patients with NS-ID (25 abnormal karyotypes out of 389 NS-ID cases). Autosomal structural abnormalities were the largest proportion of chromosomal aberrations. Conclusion: The high rate of chromosomal abnormalities found in the Moroccan patients studied demonstrates the capital importance of cytogenetic evaluation in patients who show ID or any clinical development abnormality.

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

Summary In in vitro fertilisation (IVF), sperm preparation as critical part and influencing the sperm quality is especially dependent on the chosen technique itself and incubation parameters including temperature and CO2. In this study, we compared firstly density-gradient centrifugation technique (DGC) to the adapted DGC using the sperm pellet of 80% fraction (DGC/80P) in order to improve the sperm yield. Secondly, this study led to evaluate different sperm incubation conditions based on temperature effect (room temperature (RT = 23°C) versus 35°C) and in the other hand, with or without 5% CO2 during 24 hrs. Based on evaluating sperm conventional parameters and the DNA damage using TUNEL assay, our result showed that DGC/80P increased sperm quality compared to DGC with 25% of improvement. For temperature incubation effect after 24 hrs, 35°C increased the DNA damage and decreased the sperm quality while RT could improve sperm motility by 38%. Moreover, the sperm incubation with 5% CO2 after 24 hrs realised a negative impact on sperm parameters and its DNA damage. Indeed, for current IVF practice, a good sperm quality can be maintained for several hours at room temperature, while the sperm preparation is processed using the DGC/80P without CO2.

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.