Pyridinylimidazole p38 mitogen-activated protein kinase inhibitors block intracellular Toxoplasma gondii replication

Toxoplasma gondii is a medically important, obligate intracellular parasite. Little is known regarding factors that regulate its replication within cells. Such knowledge would further understanding of T. gondii pathogenesis, and might lead to novel therapeutic strategies. Mitogen-activated protein kinases (MAPKs) govern diverse cellular processes including proliferation and differentiation. We now show that treatment of T. gondii-infected cells with SB203580 or SB202190, substituted pyridinylimidazoles that are potent inhibitors of human p38 MAPK, inhibits intracellular T. gondii replication. Several independent experimental approaches suggest that the anti-proliferative effects of pyridinylimidazoles depend on direct action on tachyzoites, not the host cell: (i) selective inhibition of host p38 MAPK using recombinant adenoviruses had little effect on tachyzoite replication, (ii) pyridinylimidazole-treated tachyzoites developed abnormal morphology suggesting defective parasite division, and (iii) pyridinylimidazole-resistant mutant tachyzoites were developed through culture in progressively higher drug concentrations. We hypothesise that pyridinylimidazoles target a human p38 MAPK homologue in tachyzoites that regulates their replication. Phylogenetic data suggest that T. gondii likely encodes a p38 MAPK homologue, but such a homologue is absent from the incomplete Toxoplasma genomic data base. As all eukaryotic pathogens, including agents of malaria, leishmaniasis and trypanosomiasis encode endogenous MAPKs, drugs inhibiting endogenous MAPK activation may represent a novel, potentially broadly-acting class of anti-parasitic agents. Pyridinylimidazoles also represent tools to elucidate factors governing intracellular tachyzoite replication.     

Thiocyanate effects on thyroid function of weaned mice

The aim of this study was to determine the effects of thiocyanate on thyroid function in weaned mice. At this developmental period, induction and reversibility of thiocyanate effects have not yet been studied. In the present work, adult female mice were given thiocyanate [SCN(-) (1 g L(-1))] in their drinking water from the 15(th) day of pregnancy until either the 25(th) (group B) or the 15(th) day (group C) after parturition. During five days after weaning, water and food consumptions of treated mice (group B) were 42.2+/-1.2% and 56.4+/-0.9%, respectively, less than those of the controls (group A). On the sacrifice day (the 25(th) day after birth), body weight, thyroid iodine content and thyroid hormone levels (FT(4) and FT(3)) decreased by 10.4+/-3.0%, 40.6+/-2.3%; 18.7+/-2.3% and 18.1+/-1.3%, respectively. Plasma TSH increased by 30.6+/-1.7% along with the hypertrophy of thyroid glands (52.6+/-3.1%). We have observed a hypertrophy of follicle cells and a decrease in colloid volume within histological slides. After SCN(-) withdrawal (group C), partial or total recovery were noted in all parameters studied. We concluded that hypothyroidism effects added to the weaning event affected greatly thyroid function and behaviour of mice; these would be largely reversed by withdrawing thiocyanate treatment for a period of ten days.     

Molecular diversity analysis and bacterial population dynamics of an adapted seawater microbiota during the degradation of Tunisian zarzatine oil

The indigenous microbiota of polluted coastal seawater in Tunisia was enriched by increasing the concentration of zarzatine crude oil. The resulting adapted microbiota was incubated with zarzatine crude oil as the only carbon and energy source. Crude oil biodegradation capacity and bacterial population dynamics of the microbiota were evaluated every week for 28 days (day 7, day 14, day 21, and day 28). Results show that the percentage of petroleum degradation was 23.9, 32.1, 65.3, and 77.8%, respectively. At day 28, non-aromatic and aromatic hydrocarbon degradation rates reached 92.6 and 68.7%, respectively. Bacterial composition of the adapted microflora was analysed by 16S rRNA gene cloning and sequencing, using total genomic DNA extracted from the adapted microflora at days 0, 7, 14, 21, and 28. Five clone libraries were constructed and a total of 430 sequences were generated and grouped into OTUs using the ARB software package. Phylogenetic analysis of the adapted microbiota shows the presence of four phylogenetic groups: Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. Diversity indices show a clear decrease in bacterial diversity of the adapted microflora according to the incubation time. The Proteobacteria are the most predominant (>80%) at day 7, day 14 and day 21 but not at day 28 for which the microbiota was reduced to only one OTU affiliated with the genus Kocuria of the Actinobacteria. This study shows that the degradation of zarzatine crude oil components depends on the activity of a specialized and dynamic seawater consortium composed of different phylogenetic taxa depending on the substrate complexity.     

Glucosylceramide synthesis inhibition affects cell cycle progression,membrane trafficking,and stage differentiation in Giardia lamblia

Synthesis of glucosylceramide via glucosylceramide synthase (GCS) is a crucial event in higher eukaryotes, both for the production of complex glycosphingolipids and for regulating cellular levels of ceramide, a potent antiproliferative second messenger. In this study, we explored the dependence of the early branching eukaryote Giardia lamblia on GCS activity. Biochemical analyses revealed that the parasite has a GCS located in endoplasmic reticulum (ER) membranes that is active in proliferating and encysting trophozoites. Pharmacological inhibition of GCS induced aberrant cell division, characterized by arrest of cytokinesis, incomplete cleavage furrow formation, and consequent block of replication. Importantly, we showed that increased ceramide levels were responsible for the cytokinesis arrest. In addition, GCS inhibition resulted in prominent ultrastructural abnormalities, including accumulation of cytosolic vesicles, enlarged lysosomes, and clathrin disorganization. Moreover, anterograde trafficking of the encystations-specific protein CWP1 was severely compromised and resulted in inhibition of stage differentiation. Our results reveal novel aspects of lipid metabolism in G. lamblia and specifically highlight the vital role of GCS in regulating cell cycle progression, membrane trafficking events, and stage differentiation in this parasite. In addition, we identified ceramide as a potent bioactive molecule, underscoring the universal conservation of ceramide signaling in eukaryotes.     

Salinity gradient and nutrients as major structuring factors of the phytoplankton communities in salt marshes

This study investigated the major environmental factors structuring, for a year, phytoplankton assemblages in the Sfax saltern (Tunisia): salinity and nutrients. A STATICO analysis based on 11 environmental variables and the abundances of 64 phytoplanktonic species was conducted. STATICO is used to analyze the stable part of the relationships between the environment and species, and then to determine how these relationships change over time. The analysis confirmed that the salinity gradient had a considerable influence on the composition of the phytoplanktonic communities. Bacillariophyceae and Dinophyceae dominated in the least salty ponds, whereas Chlorophyceae and Cyanophyceae dominated in the saltiest ponds, in accordance with the halotolerance level estimated for each species by calculating the optimum salinity and salt tolerance. Nitrogen (N) and phosphorus (P) ions could have a secondary influence on the phytoplankton distribution and its dynamics. Dinophyceae seem to be favored by high ammonium (NH₄ ⁺) concentrations, whereas diatoms seem to be favored by high orthophosphates (PO₄ ^3?) and nitrates (NO₃ ^?) values. The Chlorophyceae Dunalliela salina thrived in the saltiest ponds when the NO₃ ^? concentrations increased.     

Side effects of low serum lithium concentrations on renal, thyroid, and sexual functions in male and female rats

The present study, carried out in rats, is a contribution to explore physiological mechanisms underlying lithium toxicity. Male and female mature rats were divided into three groups and fed on commercial pellets: group (C) was control, group (Li1) was given 2000 mg lithium carbonate/kg of food, and group (Li2) was given 4000 mg lithium carbonate/kg of food. If we take into account the BW of the rats and the quantity of food they eat every day, we can estimate that the quantities of lithium carbonate ingested per day and kilogram of BW are, respectively, for the groups Li1 and Li2, of 212 mg (5,738 mmol Li) and 323 mg (8,742 mmol Li) for the males, and about 190 mg (5,142 mmol Li) and 289 mg (7,822 mmol Li) for the females. After 7, 14, 21 and 28 days, serum concentrations of lithium, creatinine, free triiodothyronine (FT3) and thyroxine (FT4), testosterone and estradiol were measured. Attention was also paid to growth rate and a histological examination of testes or vaginal mucosa was carried out. In treated rats, a dose-dependent loss of appetite and a decrease in growth rate were observed together with polydipsia, polyuria, and diarrhoea. Lithium serum concentrations were found to increase from 0.44 mM (day 7) to 1.34 mM (day 28) in Li1 rats and from 0.66 to 1.45 mM (day 14) in Li2 rats. Treatment was stopped at day 14 in Li2 rats because of a high mortality. The significant increase of creatinine that appeared, respectively, at day 7 and 14 in Li2 and Li1 rats shows that serum lithium concentrations ranging from 0.62 to 0.75 mM were able to induce renal insufficiency, secondarily leading to a time-dependent rise in lithium serum concentrations. A significant decrease of serum thyroxine (FT4) and triiodothyronine (FT3) levels was observed for lithium concentrations ranging from: 0.66 to 0.75 mmol l(-1) (Li2 rats) to 1.27 mmol l(-1) (Li1 rats). This effect was more pronounced for FT3, suggesting a defect of FT4/FT3 conversion. Under lithium treatment, the testosterone level decreased and spermatogenesis was stopped. By contrast, in treated female rats, estradiol level was found to be increased in a dose-dependent manner and animals were blocked in the diestrus phase at day 28. These results show that lithium can rapidly induce toxic effects in the rat at concentrations used for the treatment of bipolar disorders in human.     

Impairment of Spermatogenesis in Rats by Mercuric Chloride: Involvement of Low 17β-Estradiol Level in Induction of Acute Oxidative Stress

Mercuric chloride (HgCl₂) has been shown to affect the male reproductive organs, and oxidative stress has been linked with hypospermatogenesis and with male infertility. However, the specific mode of impairment of spermatogenesis during HgCl₂ exposure has not yet been clarified fully. Because of the involvement of 17β-estradiol (E2) in the male reproductive tract and its putative role on spermatogenesis, the present study aimed to investigate the possibility that HgCl₂-induced oxidative stress-mediated modulation of the E2 level exerts adverse effects on testicular steroidogenic and gametogenic activities. HgCl₂ treatment at 50 and 100 ppm for 90 days by continuous oral administration in the drink water resulted in significant dose-dependent fashion decrease in serum and testicular E₂ levels and an increase in testicular testosterone levels in dose-dependent manner, without statistical alteration in serum testosterone level among HgCl₂ exposed groups compared to the control. Cauda epididymal sperm count and motility were decreased significantly (p < 0.01), in a dose-dependent manner, in the HgCl₂-treated groups, and qualitative examination revealed inhibition of spermatogenesis and the preferential loss of maturing and elongated spermatids. The seminiferous tubules were dilated in treated animals. When compared to the control, increase in lipid peroxidation due to toxic effects of HgCl2 was accompanied by significant reduction (p < 0.01) in antioxidant enzymes activities, superoxide dismutase, catalase, and glutathione peroxidase of testes, implicating the presence of oxidative tissue damage. Furthermore, these tissue injuries caused functional impairment as evidenced with testicular elevated activity of lactate dehydrogenase. Unless oxidative stress can lead to cancer development, testis’ tumor markers as beta human chorionic gonadotropin and alpha-fetoprotein levels have shown no significant differences in the HgCl₂-exposed group compared with respect to the control. Large quantities of metal accumulated in the testis tissue are in agreement with the testis-activity failure verified in this tissue. These findings suggest that a decrease in E2 level after mercury exposure may render testis more susceptible to oxidative damage leading to its functional inactivation, thus providing new dimension to mechanisms underlying heavy metal-induced male infertility.     

Impact du thé vert “Camellia sinensis” sur les effets du vanadium sur la croissance et l’appareil génital du ratWistar mâle

Some heavy metals are known to exert harmful effects by generating an oxidative stress which, in turn, can affect the sexual and reproductive functions of male animals. The addition of antioxidants to the diet could decrease the cytotoxic effect related to oxidative stress (in the presence of heavy metals as food or water contaminants). As a contribution to this problem, the protective effect ofCamellia sinensis green tea, which is know to be rich in antioxidant compounds (polyphenols, etc.), was studied in vanadium-treated adult male rats, with particular attention to growth and genital tract function. White male Wistar rats were given ammonium metavanadate in drinking water (0.46 g/L) for 90 days One group of animals received green tea supplement in drinking water and the control group did not. Chronic vanadium intoxication (without green tea supplement) induced a low growth rate and relative atrophy of the testes, epididymis, prostate and seminal vesicles. Motility and number of spermatozoa were also decreased. Histological examination of the testes revealed atrophy of the seminiferous tubules and defects of spermatogenesis leading to the absence of spermatozoa in 50% of seminiferous tubules. Blood testosterone levels, evaluated by radioimmunoassay, were also decreased from day 2 to day 20. In control animals, these levels were 0.717±0.107 ng/ml; 4.366±0.666 ng/ml and 1.979±0.42 ng/ml on day 2, day 10 and day 20, respectively. After vanadium treatment, they were reduced to 0.043±0.012 ng/ml, 2.494±0.17 ng/ml and 1.086±0.53 ng/ml, respectively, at the same periods. These morphological, histological and functional disorders mostly occured during the first phase of the intoxication period (day 2 to day 10) and were subsequently attenuated, indicating adaptation to the poisoning. In rats receiving green tea, vanadium ingestion did not modify growth rate compared to control animals. Very minor changes were observed in the genital tract. Testicular atrophy and absence of spermatozoa were observed in only some seminiferous tubules. Our results underscore the protective effect of green tea on vanadium poisoning. Polyphenols, which are abundant in green tea, are known to chelate iron. It is proposed that polyphenols may also form insoluble complexes with vanadium, allowing it to be eliminated in the feces. This could explain the decreased effects of vanadium poisoning under our experimental conditions.