Allethrin Promotes Apoptosis and Autophagy Associated with the Oxidative Stress-Related PI3K/AKT/mTOR Signaling Pathway in Developing Rat Ovaries.

The increased concern regarding the reduction in female fertility and the impressive numbers of women undergoing fertility treatment support the existence of environmental factors beyond inappropriate programming of developing ovaries. Among these factors are pyrethroids, which are currently some of the most commonly used pesticides worldwide. The present study was performed to investigate the developmental effects of the pyrethroid-based insecticide allethrin on ovarian function in rat offspring in adulthood. We mainly focused on the roles of oxidative stress, apoptosis, autophagy and the related pathways in ovarian injury. Thirty-day-old Wistar albino female rats were intragastrically administered 0 (control), 34.2 or 68.5 mg/kg body weight allethrin after breeding from Day 6 of pregnancy until delivery. We found that allethrin-induced ovarian histopathological damage was accompanied by elevations in oxidative stress and apoptosis. Interestingly, the number of autophagosomes in allethrin-treated ovaries was higher, and this increase was correlated with the upregulated expression of genes and proteins related to the autophagic marker LC-3. Furthermore, allethrin downregulated the expression of PI3K, AKT and mTOR in allethrin-treated ovaries compared with control ovaries. Taken together, the findings of this study suggest that exposure to the pyrethroid-based insecticide allethrin adversely affects both the follicle structure and function in rat offspring during adulthood. Specifically, allethrin can induce excessive oxidative stress and defective autophagy-related apoptosis, probably through inactivation of the PI3K/AKT/mTOR signaling pathway, and these effects may contribute to ovarian dysfunction and impaired fertility in female offspring.

Permethrin induced arterial retention of native and oxidized LDL in rats by promoting inflammation, oxidative stress and affecting LDL receptors, and collagen genes.

This study is the first to examine the possible mechanism by which long-term exposure to permethrin (PER) can promote arterial retention of proatherogenic lipid and lipoproteins and related vascular dysfunction in rats. Experimental animals were administered two doses of oral PER, PER-1 (2.5 mg/kg/bw) and PER-2 (5 mg/kg/bw), for 90 consecutive days. The results indicated that both PER-1 and PER-2 increased plasmatic and aortic total cholesterol, low-density lipoprotein cholesterol (LDL-C), apo B-100, and oxidized LDL together with arterial scavenger LDL receptors (CD36) but markedly reduced plasmatic and hepatic high-density lipoprotein cholesterol and native LDL receptors in aortic and hepatic tissue. The levels of malondialdehyde, protein carbonyl, and reactive oxygen species were significantly higher, and glutathione content as well as catalase, superoxide dismutase, and glutathione peroxidase activities were suppressed in the aorta of the PER-1 and PER-2 groups. The arterial oxidative damage possibly caused by PER was clearly demonstrated by hematoxylin and eosin histological analysis. Moreover, PER treatment aggravated the inflammatory responses through enhancement of the production of proinflammatory cytokines (tumor necrosis factor-α, interleukin-2, and interleukin-6) in both plasma and aorta. Furthermore, PER-1 and PER-2 potentiated the dysregulation of the aortic extracellular matrix (ECM) content by increasing mRNA activation of collagens I and III. The abundant histological collagen deposition observed in the media and adventitia of intoxicated rats using Masson's trichrome staining corroborates the observed change in ECM. These data showed that oxidative stress related to PER exposure increases the arterial accumulation of lipoprotein biomarkers, likely by actions on both LDL and CD36 receptors, together with the disruption of the aortic ECM.

HPLC-ESI-QTOF-MS/MS profiling and therapeutic effects of Schinus terebinthifolius and Schinus molle fruits: investigation of their antioxidant, antidiabetic, anti-inflammatory and antinociceptive properties.

The aim of the current work was to study the phytochemical variability among Schinus terebinthifolius (STE) and Schinus molle (SME) fruit extracts. The in vitro antioxidant, antihemolytic, antidiabetic, and macromolecule damage protective activities, as well as, the in vivo anti-inflammatory and antinociceptive capacities were assessed. Using the HPLC-ESI-QTOF/MS analysis, the chemical profile of fruit extract varied between S. terebinthifolius (30 compounds) and S. molle (16 compounds). The major compound was masazino-flavanone (5774.98 and 1177.65 µg/g sample for STE and SME, respectively). The investigations highlighted significant antioxidant proprieties when using ABTS radical (IC50; 0.12 and 0.14 mg/ml for STE and SME, respectively), superoxide (IC50; 0.17 and 0.22 mg/ml for STE and SME, respectively) and hydrogen peroxide (IC50; 014 and 0.17 mg/ml for STE and SME, respectively). In addition, STE and SME proved preventive effects against H2O2-induced hemolysis (IC50; 0.22 and 0.14 mg/ml for STE and SME, respectively). The in vitro antidiabetic effect revealed that STE and SME exhibited important inhibitory effects against α-amylase (IC50; 0.13 and 0.19 mg/ml for STE and SME, respectively) and α-glycosidase (IC50; 0.21 and 0.18 mg/ml for STE and SME, respectively) when compared with acarbose. Furthermore, the extracts showed potent inhibitory activity against AAPH-induced plasmid DNA damage, and protein oxidation. In vivo study revealed that STE and SME presented interesting antinociceptive and anti-inflammatory capacities. All observed effects highlighted the potential application of Schinus fruit extract in food and pharmaceutical industries against ROS-induced damage.

Early-Life Exposure to the Mycotoxin Fumonisin B1 and Developmental Programming of the Ovary of the Offspring: The Possible Role of Autophagy in Fertility Recovery.

Mycotoxins are produced by more than one hundred fungi and produce secondary metabolites that contaminate various agricultural commodities, especially rice and corn. Their presence in the food chain is considered a serious problem worldwide. In recent years, a link between exposure to mycotoxins and impaired fertility has been suggested. Consequently, it has become vital to investigate the interactive effects of these mycotoxins on ovarian function. In this study, we investigated the intergenerational effects of the mycotoxin fumonisin B1 (FB1) on ovarian structure and function. Virgin Wistar albino female rats were separated into control and FB1 treatment groups and examined from day 6 of pregnancy until delivery (20 and 50 mg/kg b.w./day). The obtained female rats of the first (F1) and second generations (F2) were euthanized at 4 weeks of age, and ovary samples were collected. We found that the ovary weight index increased with the high dose of the treatment (50 mg/kg b.w./day) among both F1 and F2, in a manner similar to that observed in polycystic ovary syndrome. As expected, FB1 at a high dose (50 mg/kg b.w.) reduced the number of primordial follicles in F1 and F2, leading to an accelerated age-related decline in reproductive capacity. Moreover, it reduced the fertility rate among the F1 female rats by affecting follicle growth and development, as the number of secondary and tertiary follicles decreased. Histopathological changes were evidenced by the altered structures of most of the growing follicle oocytes, as revealed by a thinning irregular zona pellucida and pyknosis in granulosa cells. These findings are concomitant with steroidogenesis- and folliculogenesis-related gene expression, as evidenced by the decrease in CYP19 activity and estrogen receptor beta (ESR2) gene expression. Additionally, GDF-9 mRNA levels were significantly decreased, and IGF-1 mRNA levels were significantly increased. However, the results from the ovaries of the F2 treatment groups were different and unexpected. While there was no significant variation in CYP19 activity compared to the control, the ESR2 significantly increased, leading to stereological and histopathological changes similar to those of the control, except for some altered follicles. The hallmark histological feature was the appearance of vacuolar structures within the oocyte and between granulosa cell layers. Interestingly, the autophagic marker LC3 was significantly increased in the F2 offspring, whereas this protein was significantly decreased in the F1 offspring. Therefore, we suggest that the promotion of autophagy in the ovaries of the F2 offspring may be considered a recovery mechanism from the effect of prenatal FB1 exposure. Thus, autophagy corrected the effect of FB1 during the early life of the F1 female rats, leading to F2 offspring with ovarian structure and function similar to those of the control. However, the offspring, treated female rats may experience early ovarian aging because their ovarian pool was affected.

Ephedra alata Subsp. Alenda as a Novel Source of Bioactive Phytochemicals: Characterization Based on the Mass Spectrometry and Profiling of Antioxidant and Anti-Inflammatory Properties.

The aim of the present study was to examine, for the first time, the phytochemical content of Ephedra alata pulp extract (EAP) and explore its antioxidant and anti-inflammatory capacities. High-performance liquid chromatography-electrospray ionization-quadrupole-time-of-flight mass spectrometry (HPLC-ESI-QTOF/MS) was used for phytochemical analysis and three in vitro antioxidant assays together with three in vitro anti-inflammatory tests were used for the assessment of biological activity. The HPLC-ESI-QTOF/MS analysis revealed the presence of 42 metabolites, including flavonoids, sphingolipides, fatty acids, ephedrine derivatives, and amino acid derivatives. In vitro findings revealed that EAP has interesting 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide, and ferrous ion chelating capacities (IC50 values were 0.57 mg/mL, 0.55 mg/mL, and 0.51 mg/mL for DPPH, superoxide radical, and ferrous ion, respectively). Furthermore, EAP showed a noticeable anti-inflammatory ability by inhibiting the two cyclooxygenase isoforms, COX-1 and COX-2 (IC50 of 59.1 and 58.8 µg/mL for COX-1 and COX-2, respectively), preventing protein denaturation (IC50 = 0.51 mg/mL), and protecting membrane stabilization (IC50 = 0.53 mg/mL). The results highlighted the use of Ephedra alata pulp as a potential source of natural compounds with therapeutic effects for the management of inflammatory disorders.

Maternal Exposure to Acephate Caused Nephrotoxicity in Adult Offspring Rats Mediated by Excessive Autophagy Activation, Oxidative Stress Induction, and Altered Epithelial Sodium Channel and Na+/K+-ATPase Gene Expression.

This study examined how maternal exposure to acephate-an organophosphate-based insecticide-affected the renal development in rat offspring during adulthood. Virgin female Wistar rats were randomly allocated to three groups: group 1 (control) received sterile water; groups 2 and 3 were intragastrically exposed to low (14 mg/kg) and high (28 mg/kg) doses of acephate from day 6 of pregnancy until delivery, respectively. Further, the offspring of the adult female rats were euthanized in postnatal week 8. Compared with the controls, the adult rat offspring with exposure to low and high doses of acephate exhibited elevated plasma creatinine and blood urea nitrogen levels. Additionally, immunofluorescence analysis revealed the upregulation of autophagic marker genes (Beclin-1 and LC-3) in the acephate-treated rat offspring, thereby suggesting the induction of an autophagic mechanism. Notably, the increased malondialdehyde level, decreased glutathione level, and decreased superoxide dismutase and catalase activities confirmed the ability of acephate to induce oxidative stress and apoptosis in the kidneys of the rat offspring. This may explain the renal histopathological injury detected using hematoxylin and eosin staining. Furthermore, a reverse transcription polymerase chain reaction revealed that the mRNA expression levels of the Na+/K+-ATPase and the epithelial sodium channel (ENaC) genes were significantly higher in the kidney of female offspring than that of controls owing to acephate toxicity. However, there was no significant effect of acephate on the expression of NHE3 in the treatment group compared with the control group. Overall, the present findings suggest that oxidative stress caused by prenatal exposure to acephate causes nephrotoxicity and histopathological alterations in adult rat offspring, likely by actions on renal ENaC and Na+/K+-ATPase genes as well as the autophagic markers Beclin-1 and LC-3.

Effects of an Endocrine Disruptor Triclosan on Ruditapes decussatus: Multimarker and Histological Approaches.

The aim of this work was to study the ecotoxicological effects of an endocrine disruptor triclosan on the clam Ruditapes decussatus. The bivalves were exposed to three concentrations of this biocide (C1 = 100 ng/L, C2 = 200 ng/L and C3 = 500 ng/L) for three and seven days. The impact was assessed at the gills and digestive glands, through activities of an antioxidant defense biomarker (Gluthatione S-Transferase, GST), a damage biomarker (Malondialdehyde, MDA), and a neurotoxicity biomarker (Acetylcholinesterase, AChE). Furthermore, histological traits were approached in different organs to evaluate any possible alteration induced by triclosan. It appears from this study that both gills and digestive glands responded discernibly to triclosan and effects were concentration-dependent. The stressed clams showed a significant increase in their GST and MDA activities in gills and digestive glands compared to controls for both time slots considered. In turn, the AChE activity was clearly inhibited in both organs in a time dependent way. The histological study made it possible to observe several structural pathologies caused by triclosan in the gills and the digestive gland. These alterations consisted mainly of inflammatory reactions, malformations of the lamellae and fusion of the gill filaments, degeneration of the connective tissue, and the erosion of the gill cilia with the appearance of certain severe alterations (cell necrosis and apoptosis), which can thus cause a malfunction of the gills and eventually lead to a reduction in oxygen consumption and a disruption of the osmoregulation for bivalves. Alterations in the digestive gland have also been detected, mainly by epithelial alterations, thinning of the tubules, and alteration of the basal cell membrane which can impair the ability of clams to absorb food. At germinal cells, several damages were observed in the oocytes which probably disturbed the reproductive function and the fertility of the clams. The damages observed in female gonads were caused by the cytolysis of a large number of oocytes through autophagy and necrosis at 200 ng triclosan/L. Moreover, at 500 ng triclosan/L, hemocytic infiltration was observed in acini and apoptotic bodies reflected in the fragmentation of more than 90% of oocytes.

Morphological Redescription and Phylogenetical Position of Ceratomyxa truncata Thelohan (1895) and Coccomyxa morovi Léger and Hesse, 1907 (Myxozoa: Myxosporea) Infecting the Gall Bladder of Sardina pilchardus (Walbaum) from Tunisian Coast.

PURPOSE: Two myxosporean species have been, so far, independently reported from the gallbladder of the European pilchard, Sardina pilchardus (Walbaum) (synonym Clupea pilchardus) in the Northern shore of the Mediterranean Sea; Ceratomyxa truncata Thélohan, and Coccomyxa morovi Léger and Hesse, 1907. The two species were described with incomplete morphological data and based only on line drawings of their mature myxospores. METHODS: During a parasitological survey in the Southern shores of the Mediterranean coast in the gulf of Gabès off Tunisia, two coelozoic myxosporean species were found in the European pilchard and described using morphological and molecular phylogenetic tools. Morphological characterization was based on the mature myxospore study and some vegetative stages. The SSU rDNA sequences were performed for molecular and phylogenetic study. RESULTS: The most frequently encountered species belongs to the genus Ceratomyxa Thelohan, 1892. The second species belongs to the genus Coccomyxa. Morphological examinations, allowed us to match these two recorded species with Ceratomyxa truncata and Coccomyxa morovi, respectively, as previously described in the same host species referring to the original manuscripts instead of some morphological differences. Molecular analyses based on the partial SSU rDNA sequences did not much with any of the previously reported myxozoan sequences. Phylogenetic analysis positioned C. truncate in a well-supported clade including Ceratomyxa ssp. from Mediterranean Sea, while C. morovi was positioned on the basis of the subclade grouping all Coccomyxidae species. CONCLUSION: We provided herein a first morphological redescription of Ceratomyxa truncata and Coccomyxa morovi parasite of Sardina pilchardus from the Southern shores of the Mediterranean Sea and we successfully obtained the SSU rDNA sequences of these two species and positioned them in the phylogenetic tree.

Fetal programming: in utero exposure to acrylamide leads to intergenerational disrupted ovarian function and accelerated ovarian aging.

In this study we investigated the effects of multigenerational exposures to acrylamide (ACR) on ovarian function. Fifty-day-old Wistar albino female rats were divided into the control and ACR-treated groups (2.5, 10, and 20 mg/kg/day) from day 6 of pregnancy until delivery. The obtained females of the first (AF1) and second generation (AF2) were euthanized at 4 weeks of age, and plasma and ovary samples were collected. We found that in utero multigenerational exposure to ACR reduced fertility and ovarian function in AF1 through inducing histopathological changes as evidenced by the appearance of cysts and degenerating follicles, oocyte vacuolization, and pyknosis in granulosa cells. TMR red positive cells confirmed by TUNEL assay were mostly detected in the stroma of the treated groups. Estradiol and IGF-1 concentrations significantly decreased as a result of decreased CYP19 gene and its protein expression. However, ACR exposure in AF2 led to early ovarian aging as evidenced by high estradiol and progesterone levels among all treated groups compared to control group, corresponding to the upregulation of the CYP19 gene and protein expression. The apoptotic cells of the stroma were greatly detected compared to that in the control group, whereas no significant difference was reported in ESR1 and ESR2 gene expression. This study confirms the developmental adverse effects of ACR on ovarian function and fertility in at least two consecutive generations. It emphasizes the need for more effective strategies during pregnancy, such as eating healthy foods and avoiding consumption of ACR-rich products, including fried foods and coffee.

Physiological Responses of the Bivalves Mytilus galloprovincialis and Ruditapes decussatus Following Exposure to Phenanthrene: Toxicokinetics, Dynamics and Biomarkers Study.

The aim of the current study was to assess the multifaceted effects of the polycylic aromatic hydrocarbon phenanthrene, mainly used in the colouring, explosive, and pharmaceutical industries, on the physiology of two bivalve species with economic value as seafood, namely, the Mediterranean mussel Mytilus galloprovincyalis and the European clam Ruditapes decussatus. The current study assessed how the phenanthrene affected several biomarkers and biometric endpoints in both bivalves, based on an in vivo experiment in silico approach. The bivalves were exposed during four time slots (i.e., 7, 15, 21, and 28 days) to two concentrations of phenanthrene in water (50 µg/L and 100 µg/L). For the clam R. decussatus, an additional contamination of sediment was applied due their typical benthic lifestyle (50 µg/kg and 100 µg/kg). The phenanthrene significantly reduced the ability of bivalves to tolerate desiccation and their Median Lethal Time, and also inhibited the activity of the enzyme acetylcholinesterase in a time-dependent manner. The activity of catalase indicated that bivalves also experienced oxidative stress during the first 21 days of the experiment. The significant decline in catalase activity observed during the last week of the experiment for the mussel M. galloprovincyalis supported a depletion of enzymes caused by the phenanthrene. The phenanthrene has also toxicokinetic and toxicodynamic properties, as assessed by the in silico approach. Overall, the results obtained suggest that the bivalves Ruditapes decussatus and M. galloprovincyalis can be used as a sentinel species in monitoring studies to assess the environmental impact of phenanthene in marine ecosystems. The significance of our findings is based on the fact that in ecotoxicology, little is known about the chronic effects, the simultaneous use of multiple species as bioindicators, and the interactions molecular modelling.

Schinus terebinthifolius fruits intake ameliorates metabolic disorders, inflammation, oxidative stress, and related vascular dysfunction, in atherogenic diet-induced obese rats. Insight of their chemical characterization using HPLC-ESI-QTOF-MS/MS.

ETHNOPHARMACOLOGY RELEVANCE: Schinus terebinthifolius is traditionally used for its anti inflammatory capacity, and indicated as a cardioprotective agent, whereas, its preventive effect against atherogenic diet fed (AD) induced metabolic disorders and the underlying mechanisms has not yet been explored. AIM OF THE STUDY: This study was undertaken to investigate the ameliorative role of Schinus terebinthifolius fruits extract (STFE) against cardiovascular problem, oxidative and inflammatory status related to obesity in rats fed an atherogenic diet. MATERIALS AND METHODS: The metabolites profile in STFE was evaluated using HPLC-DAD-ESI-QTOF-MS/MS analysis. In Wistar rats, atherogenic diet was added for 9 weeks to induce lipid accumulation simultaneously with STFE (50 mg/kg b. w) or saline treatment. Biochemical, oxidant, and inflammatory criteria together with hepatic and arterial integrity examination were assessed. RESULTS: A total of thirty three metabolites were identified using HPLC-DAD-ESI-QTOF-MS, among them masazino-flavanone was the major compound (2645.50 µg/g DW). The results indicated that STFE supplementation during 9 weeks (50 mg/kg b. w.) significantly attenuated the altered lipid profile by decreasing the levels of TC, TG, LDL-C and increasing the HDL-C content both in plasma and liver, when compared with the AD-group. The histological analysis using ORO staining revealed a decrease in the lipid droplet deposit in the cytoplasm of hepatocytes of STFE + AD group. The addition of STFE could improve the glycemic status of AD-treated rats by decreasing the glucose and insulin secretion, and ameliorating the hepatic glycogen synthesis. The harmful effects of atherogenic diet on hepatic oxidative stress indicators (MDA, PC, GSH, SOD, CAT, and GPx), biochemical markers (AST, ALT, LDH and ALP), and liver function, were found to be decreased by the addition of STFE. Moreover, the reduction of inflammatory markers (CRP, IL-6 and TNF-α), associated to alleviating of aortic oxidative stress and integrity, highlighted the positive anti-atherogenic effect of STFE. CONCLUSION: Overall, the pleiotropic protective effect observed with S. terebinthifolius fruits might be related to the presence of various bioactive compounds.

Multidirectional insights on polysaccharides from Schinus terebinthifolius and Schinus molle fruits: Physicochemical and functional profiles, in vitro antioxidant, anti-genotoxicity, antidiabetic, and antihemolytic capacities, and in vivo anti-inflammatory and anti-nociceptive properties.

The aim of the current study was to compare crude polysaccharides extracted from Schinus terebinthifolius Raddi (PSTF) and S. molle L. (PSMF) fruits based on their structures, physicochemical characteristics, monosaccharide composition, as well as in vitro and in vivo assays. The extraction yield of PSTF (4.26%) was higher than that of PSMF (3.56%). Remarkable variability was detected in the content of carbohydrates (80.64 ± 0.98%), protein (1.80 ± 0.28%), fat (0.04 ± 0.005%) and ash (6.32 ± 0.26%). FT-IR assay and 1H and 13C NMR spectroscopy revealed that fruits extract showed similar structural characteristics. Thin layer chromatography together with HPLC-RID analysis showed that the monosaccharide composition varied significantly between species. Both contained arabinose (40.55-42.03%) galacturonic acid (31.21-41.15%), and fucose (10.90-17.63%), but PSTF had glucose (9.13%) whereas PSMF had galactose (7.40%). Functional analyses demonstrated that samples exhibited favorable water- and oil-retention capacity, emulsifying properties, and foaming qualities. PSTF exhibited the highest antioxidant effects. Both of them showed a remarkable in vitro antidiabetic effect. PSMF highly mitigated H2O2-induced hemolysis and exhibited ~80% antihemolytic activity. The extracted polysaccharides showed potent inhibitory activity against AAPH-induced plasmid DNA damage. PSTF and PSMF revealed interesting in vivo antinociceptive and anti-inflammatory capacities.

Epidemiology of neural tube defect subtypes in Tunisia, 1991-2011.

BACKGROUND: Neural tube defects are common major congenital anomalies that result from very early disruption in the development of the brain and spinal cord. AIM OF THE STUDY: We conducted an epidemiological study to determine the impact of some feto-maternal characteristics in the occurrence of NTD subtypes. METHODS: Characteristics and outcomes of births with NTD and pregnancy characteristics of mothers over a period of twenty years (1991-2011) were recorded in the medical chart. RESULTS: From 1991 through 2011, 769 stillborns with NTD were delivered, yielding a prevalence of 2.02/10,000. The increase in NTD prevalences over these years was statistically significant (P = 0.000). In addition, differences between prevalences of NTD subtypes over season (P = 0.003) and between genders (P < 0.001) were significant. The highest frequency was noticed in winter with 3, 7 per 10,000 births among females. The difference in fetal term between subtypes was significant (P = 0.017). The probability to have a malformed fetus with a weight less than 1500 g was three times higher in myelomeningocele than in craniorachischisis, two times higher in anencephaly and encephalocele, but two times lower than rachischisis. Mothers with one gestation were two fold higher in anencephaly than in encephalocele. Nulliparous mothers' cases were significantly more likely to have NTD than uni- or multiparous mothers. O+ mother's blood type presented a significant risk factor and was significantly less common in myelomeningocele than in rachischisis, but three times higher than in craniorachischisis. Consanguinity was present in cases with rachischisis and was two times higher than in cases with anencephaly, and three times higher than in cases with encephalocele. In this study, the results have been interpreted with caution due to analyses not being adjusted. CONCLUSION: One of the main findings of the study is that there are many differences between NTD subtypes, which suggests that there may be etiologic differences between subtypes. This suggests that, although epidemiologic studies frequently do not distinguish between NTD subtypes in analyses, they should be analyzed separately when possible.