Probiotics from kefir: Evaluating their immunostimulant and antioxidant potential in the carpet shell clam (Ruditapesdecussatus).

This study aimed to investigate the impact of incorporating kefir into the diet on biometric parameters, as well as the immune and antioxidant responses of the carpet shell clam (Ruditapes decussatus) after an experimental infection by Vibrio alginolyticus. Clams were divided into a control group and a treated group. The control group was fed on spirulina (Arthrospira platensis) alone. While, the treated group was fed on spirulina supplemented with 10% dried kefir. After 21 days, clams were immersed in a suspension of V. alginolyticus 5 × 105 CFU mL -1 for 30 min. Seven days after experimental infection, survival was 100% in both groups. The obtained results showed a slight increase in weight and condition index in clams fed with kefir-supplemented diet for 21 days compared to control clams. Regarding antioxidant responses, the treated group showed higher superoxide dismutase activity compared to the control group. However, the malondialdehyde level was lower in the treated clams than in the control. In terms of immune parameters, the treated group showed slightly elevated activities of phenoloxidase, lysozyme and alkaline phosphatase, whereas a decreased lectin activity was observed compared to the control group. The obtained results suggest that kefir enhanced both the antioxidant and immune response of infected clams.

Biological Monitoring of Occupational Exposure to Metals in Electric Steel Foundry Workers and Its Contribution to 8-Oxo-7,8-Dihydro-2'-Deoxyguanosine Levels.

In this study, the urinary concentrations of selected metals in workers from an electric steel foundry in Tunisia were assessed and compared with existing biological limit values and general population reference values. Moreover, the association between oxidative DNA damage, measured as urinary 8-oxo-7,8-dihydro-2'deoxyguanosine (8-oxodG) and co-exposure to metals and polycyclic aromatic hydrocarbons (PAHs) was evaluated. Urinary levels of 12 metals were determined by inductively coupled plasma-mass spectrometry (ICP-MS) in end-shift spot samples from 89 workers. The urinary levels of phenanthrene (U-PHE), as marker of exposure to PAHs, and 8-oxodG were also available. Median levels ranged from 0.4 µg/L (cobalt, Co, and thallium, Tl) to 895 µg/L (zinc, Zn). Only 1% of samples was above the biological limit values for Co, and up to 13.5% of samples were above limit values for Cd. From 3.4% (Co) to 72% (lead, Pb) of samples were above the reference values for the general population. Multiple linear regression models, showed that manganese (Mn), Zn, arsenic (As), barium (Ba), Tl, and Pb were significant predictors of 8-oxodG (0.012 ≤ p ≤ 0.048); U-PHE was also a significant predictor (0.003 ≤ p ≤ 0.059). The variance explained by models was low (0.11 ≤ R2 ≤ 0.17, p < 0.005), showing that metals and PAHs were minor contributors to 8-oxodG. Overall, the comparison with biological limit values showed that the study subjects were occupationally exposed to metals, with levels exceeding biological limit values only for Cd.

Oxidative Stress and Damage Biomarkers in Clam Ruditapes decussatus Exposed to a Polluted Site: The Reliable Biomonitoring Tools in Hot and Cold Seasons.

In the present study, a multi-biomarker approach was used to assess the biological effects of metal pollution in the southern lagoon of Tunis, on clam Ruditapes decussatus both in "hot" (in summer) and "cold" (in winter) seasons. Clams were collected in August 2015 and February 2016 from three sites of the lagoon and from Louza considered a reference site. The concentrations of five trace metals (cadmium, copper, iron, lead, and zinc) in the soft tissues of R. decussatus were evaluated at the sampling sites. A core of biomarkers indicative of (a) neurotoxicity (acetylcholinesterase, AChE); (b) biotransformation (glutathione S-transferase, GST); (c) oxidative stress (catalase, CAT; total glutathione peroxidase, T-GPx; total glutathione peroxidase, T-GPx; selenium-dependent glutathione peroxidase, Se-GPx; glutathione reductase, GR; superoxide dismutase, SOD) (d) lipid peroxidation (malondialdhyde, MDA level), and (e) apoptotic process (caspase 3-like, CSP3) was selected for measurements of environmental effects on the populations of clams collected from the different sampling sites. The results of metal bioaccumulation in soft tissues of Ruditapes decussatus revealed a high pollution in the South Lagoon of Tunis with spatial variation and relatively high levels at the navigation channel. Anthropogenic pollutants in the lagoon led to the activation of antioxidant defense and biotransformation enzymes to oxidative damage of the membrane and activation of apoptosis, and revealed neurotoxicity. Among this core of biomarkers, the antioxidants enzymes (CAT, SOD, GR, and GPx) were very sensitive, allowing the discrimination among sites and pointing to the navigation channel as the most impacted site in the southern lagoon of Tunis. Moreover, a significant effect of season was recorded on biomarkers responses (e.g., CAT, GR, SOD, AChE, and CSP3 activities and MDA levels) with higher levels in winter than in summer, probably influenced by the reproductive stage and food availability. Finally, the measurement of the selected core of biomarkers in the whole soft tissues of clams was considered as an integrated indicator of environmental stress. Moreover, R. decussatus proved to be a remarkable sentinel species capable to establish a reliable diagnosis of the health status of the marine environment in different areas of the southern lagoon of Tunis, both in "hot" and "cold" seasons.

Occurrence, sources and effects of polycyclic aromatic hydrocarbons in the Tunis lagoon, Tunisia: an integrated approach using multi-level biological responses in Ruditapes decussatus.

Coastal lagoons are critical ecosystems presenting a strategic economic importance, but they are subjected to potential anthropogenic impact. As part of the Tunis lagoon (Tunisia) biomonitoring study, levels, composition pattern and sources of polycyclic aromatic hydrocarbons (PAHs) in surface sediments along with their bioavailability in clam Ruditapes decussatus were investigated in polluted (S2-S4) and reference (S1) sites. In order to investigate the contamination effects at different biological levels in clams, a wide set of biomarkers, including gene expression changes, enzymatic activities disruption and histopathological alterations, was analysed. Biomarkers were integrated in a biomarker index (IBR index) to allow a global assessment of the biological response. Principal component analysis (PCA) was used for chemical and biological data integration to rank the sampling sites according to their global environmental quality. Sediment PAHs levels ranged between 144.5 and 3887.0 ng g-1 dw in the Tunis lagoon sites versus 92.6 ng g-1 dw in the reference site. The high PAH concentrations are due to anthropogenic activities around the lagoon. PAH composition profiles and diagnostic isomer ratios analysis indicated that PAHs were of both pyrolitic and petrogenic origins. Clams sampled from S2 and S3 exhibited the highest PAH contents with 2192.6 ng g-1 dw and 2371.4 ng g-1 dw, respectively. Elevated levels of tissue PAHs were associated to an increase in biotransformation and antioxidant activities, and lipid peroxidation levels along with an overexpression of different genes encoding for general stress response, mitochondrial metabolism and antioxidant defence, in addition to the emergence of severe and diverse histopathological alterations in the clams' digestive glands. IBR index was suitable for sampling sites ranking (S1 = 0 < S4 = 0.4 < S3 = 1.15 < S2 = 1.27) based on the level of PAH-induced stress in clams. PCA approach produced two components (PC1, 83.8% and PC2, 12.2%) that describe 96% of the variance in the data and thus highlighted the importance of integrating contaminants in sediments, their bioaccumulation and a battery of biomarkers of different dimensions for the assessment of global health status of coastal and lagoon areas.

A Nanocomposite Based on Reduced Graphene and Gold Nanoparticles for Highly Sensitive Electrochemical Detection of Pseudomonas aeruginosa through Its Virulence Factors.

Pyoverdine is a fluorescent siderophore produced by Pseudomonas aeruginosa that can be considered as a detectable marker in nosocomial infections. The presence of pyoverdine in water can be directly linked to the presence of the P. aeruginosa, thus being a nontoxic and low-cost marker for the detection of biological contamination. A novel platform was developed and applied for the electrochemical selective and sensitive detection of pyoverdine, based on a graphene/graphite-modified screen-printed electrode (SPE) that was electrochemically reduced and decorated with gold nanoparticles (NPs). The optimized sensor presenting higher sensitivity towards pyoverdine was successfully applied for its detection in real samples (serum, saliva, and tap water), in the presence of various interfering species. The excellent analytical performances underline the premises for an early diagnosis kit of bacterial infections based on electrochemical sensors.

Urinary 8-Oxo-7,8-Dihydro-2'-Deoxyguanosine in Tunisian Electric Steel Foundry Workers Exposed to Polycyclic Aromatic Hydrocarbons.

In this study, urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), as biomarker of oxidative DNA damage, was evaluated in Tunisian electric steel foundry workers and was associated with polycyclic aromatic hydrocarbon (PAH) exposure. Ninety-three healthy male workers were enrolled in the study; 8-oxodG was assessed by liquid chromatography-triple quadrupole mass spectrometry. Exposure to PAHs was evaluated by measuring 16 urinary PAHs (U-PAHs) and 8 monohydroxylated metabolites (OHPAHs). The median 8-oxodG level for all subjects was 3.20 µg l-1 (1.85 µg g-1 creatinine). No correlation between 8-oxodG and 1-hydroxypyrene or any other OHPAH was found. Significant linear correlations between 8-oxodG and some U-PAHs were found, particularly urinary acenaphthylene (r = 0.249), phenanthrene (r = 0.327), anthracene (r = 0.357), fluoranthene (r = 0.248), and pyrene (r = 0.244). Multiple regression analyses confirmed that urinary phenanthrene, anthracene, and naphthalene (the latter with a non-linear relationship) were predictors of 8-oxodG; job title, but not smoking, was a determinant of 8-oxodG; the variance explained by these models was up to 20%. The oxidative DNA damage assessed by urinary 8-oxodG was moderate and in the range of values reported in other occupational fields or in the general population. The results of this study indicate that the investigated biomarkers of PAH exposure were only minor contributors to urinary 8-oxodG.

Combined effects of metal contamination and abiotic parameters on biomarker responses in clam Ruditapes decussatus gills: an integrated approach in biomonitoring of Tunis lagoon.

The spatial and seasonal alteration in a battery of biomarkers responses (enzymatic activity of glutathione-S-transferase, catalase and acetylcholinesterase and lipid peroxidation) were investigated to assess the metal derived effects in clam (Ruditapes decussatus) gills, collected from Tunis lagoon (Tunisia). Trace metals (Ag, As, Cd, Cu, Hg, Mn, Ni, Pb, V and Zn) concentrations were assessed seasonally in sediments and tissues of R. decussatus from three different sites (S1, S2 and S3). Bioaccumulation factor (BAF) analysis showed a spatio-temporal variation of metal uptake rates in clams through sediments. Likewise, the multibiomarker approach enabled a time-site trend differentiation between sites with distinctive degrees of anthropogenic contamination. Site S2 was identified as the most impacted region due to the presence of different contamination sources (shipping and industrial activities). The results suggest that biomarker's seasonal variation arises from a complex interaction between environmental conditions (e.g. temperature, salinity) and probably biological factors (mainly the reproduction process) along with anthropogenic pressure. The general biological response measured with the IBR index at all the sampling sites revealed the highest metabolic stress in summer. The combined effects of metal contamination and increased temperature and salinity in summer appear to induce the highest metabolic adaptation response. The selected biomarkers provided an integrated response, which is useful for the assessment of the combined effects of metal contamination and abiotic parameters in clams and the environmental status of coastal lagoon ecosystem.

Biological Monitoring of Occupational Exposure to Polycyclic Aromatic Hydrocarbons at an Electric Steel Foundry in Tunisia.

Occupational exposures during iron and steel founding have been classified as carcinogenic to humans, and the exposure to polycyclic aromatic hydrocarbons (PAHs) in this industrial setting may contribute to cancer risk. The occupational exposure to PAHs was assessed in 93 male workers at an electric steel foundry in Tunisia by biomonitoring, with the aims of characterizing the excretion profile and investigating the influence of job title and personal characteristics on the biomarkers. Sixteen 2-6 ring unmetabolized PAHs (U-PAHs) and eight hydroxylated PAH metabolites (OHPAHs) were analyzed by gas chromatography-triple quadrupole tandem mass spectrometry and liquid chromatography triple quadrupole tandem mass spectrometry, respectively. Among U-PAHs, urinary naphthalene (U-NAP) was the most abundant compound (median level: 643ng l(-1)), followed by phenanthrene (U-PHE, 18.5ng l(-1)). Urinary benzo[a]pyrene (U-BaP) level was <0.30ng l(-1) Among OHPAHs, 2-hydroxynaphthalene (2-OHNAP) was the most abundant metabolite (2.27 µg l(-1)). Median 1-hydroxypyrene (1-OHPYR) was 0.52 µg l(-1) Significant correlations among urinary biomarkers were observed, with Pearson's r ranging from 0.177 to 0.626. 1-OHPYR was correlated to benzo[a]pyrene, but not to five- and six-rings PAHs. A multiple linear regression model showed that job title was a significant determinant for almost all U-PAHs. In particular, employees in the steel smelter workshop had higher levels of high-boiling U-PAHs and lower levels of low-boiling U-PAHs than those of workers with other job titles. Among OHPAHs, this model was significant only for naphthols and 1-hydroxyphenanthrene (1-OHPHE). Smoking status was a significant predictor for almost all biomarkers. Among all analytes, U-PHE and 1-OHPHE were the less affected by tobacco smoke, and they were significantly correlated with both low- and high-molecular-weight compounds, and their levels were related to job titles, so they could be proposed as suitable biomarkers of PAH exposure at steel foundries. Based on 1-OHPYR levels, our findings show that occupational exposure of these workers was similar to that reported in recent studies of electric steel foundry workers. The multianalytic approach is useful in revealing different exposure levels among job titles.

Transcriptomic, Biochemical, and Histopathological Responses of the Clam Ruditapes decussatus from a Metal-Contaminated Tunis Lagoon.

This study was designed to investigate the molecular (transcriptional expression), biochemical (oxidative stress and neurotoxicity), and histopathological effects of metal contamination in the gill of clams (Ruditapes decussatus) sampled from the Tunis lagoon. The concentrations of five heavy metals (Cd, Pb, Hg, Cu, and Zn) in surface sediments and their accumulation in soft tissues of R. decussatus were evaluated in three sites (Z1, Z2, and Z3). A metal contamination state of Tunis lagoon sediments was noted with spatial variations with relatively high levels at Z2. Biomarker analyses showed an increase in glutathione S-transferase and catalase activities and lipid peroxidation levels and a decrease in acetylcholinesterase activity in the studied sites. Molecular investigation showed a significant overexpression of: cytochrome c oxidase subunit I, ribosomal RNA 16S, Cu/Zn superoxide dismutase, heat shock protein 70, and metallothioneins in the three sampling sites. Moreover, our data were correlated to severe and diverse histopathological alterations in the clam gills. The principal component analysis showed that the Z2 region is more affected by metal contamination than Z1 and Z3 regions. Current field results suggest the use of several combined biomarkers at different cell levels instead of individual ones in monitoring programs.

Attenuation of endoplasmic reticulum stress and mitochondrial injury in kidney with ischemic postconditioning application and trimetazidine treatment.

BACKGROUND: Endoplasmic reticulum (ER) and mitochondria have been implicated in the pathology of renal ischemia/reperfusion (I/R). In the present study, we investigated whether the use of ischemic postconditioning (IPostC) and trimetazidine (TMZ) separately or combined could reduce ER stress and mitochondria damage after renal ischemia. METHODS: Kidneys of Wistar rats were subjected to 60-min of warm ischemia followed by 120-min of reperfusion (I/R group, n = 6), or to 6 cycles of ischemia/reperfusion (10-s each cycle) just after 60-min of warm ischemia (IPostC group, n = 6), or to i.p. injection of TMZ (3 mg/kg) 30-min before ischemia (TMZ group, n = 6), or to the combination of both treatments (IPostC+TMZ group, n = 6). The results of these experimental groups were compared to those of a sham-operated group in which rat renal pedicles were only dissected. Sodium reabsorption rate, creatinine clearance lactate deshydrogenase (LDH) activity in plasma, and concentration of malonedialdehyde (MDA) in tissue were determined. In addition, Western blot analysis was performed to identify the amounts of cytochrome c, c-JunNH2-terminal kinase (JNK), voltage-dependent anion channel (VDAC), glycogen synthase kinase 3-beta (GSK3-ß), and ER stress parameters. RESULTS: IPostC or/and TMZ significantly decreased cytolysis, oxidative stress and improved renal function in comparison to I/R group. IPostC but not TMZ significantly attenuated ER stress parameters versus I/R group. Indeed, it down-regulated the glucose-regulated protein 78 (GRP78), the activating transcription factor 4 (ATF4), the RNA activated protein kinase (PKR)-like ER kinas (PERK), the X box binding protein-1 (XBP-1) and the caspase12 protein levels. TMZ treatment significantly augmented GSK3-ß phosphorylation and reduced levels of cytochrome c and VDAC phosphorylation in comparison to IPostC application. The combination of both treatments gave a synergetic effect. It significantly improved the survival rate, attenuated cytolysis, oxidative stress and improved renal function. CONCLUSION: This study revealed that IPostC protects kidney from I/R injury by suppressing ER stress while the beneficial effects of TMZ are mediated by mitochondria protection. The combination of both treatments ameliorated functional recovery.

Ischemic preconditioning reduces endoplasmic reticulum stress and upregulates hypoxia inducible factor-1α in ischemic kidney: the role of nitric oxide.

BACKGROUND: Although recent studies indicate that renal ischemic preconditioning (IPC) protects the kidney from ischemia-reperfusion (I/R) injury, the precise protective mechanism remains unclear. In the current study, we investigated whether early IPC could upregulate hypoxia inducible transcription factor-1α (HIF-1α) expression and could reduce endoplasmic reticulum (ER) stress after renal I/R and whether pharmacological inhibition of nitric oxide (NO) production would abolish these protective effects. METHODS: Kidneys of Wistar rats were subjected to 60 min of warm ischemia followed by 120 min of reperfusion (I/R group), or to 2 preceding cycles of 5 min ischemia and 5 min reperfusion (IPC group), or to intravenously injection of NG-nitro-L-arginine methylester (L-NAME, 5 mg/kg) 5 min before IPC (L-NAME+IPC group). The results of these experimental groups were compared to those of a sham-operated group. Sodium reabsorption rate, creatinine clearance, plasma lactate dehydrogenase (LDH) activity, tissues concentrations of malonedialdehyde (MDA), HIF-1α and nitrite/nitrate were determined. In addition, Western blot analyses were performed to identify the amounts of Akt, endothelial nitric oxide synthase (eNOS) and ER stress parameters. RESULTS: IPC decreased cytolysis, lipid peroxidation and improved renal function. Parallelly, IPC enhanced Akt phosphorylation, eNOS, nitrite/nitrate and HIF-1α levels as compared to I/R group. Moreover, our results showed that IPC increased the relative amounts of glucose-regulated protein 78 (GRP78) and decreased those of RNA activated protein kinase (PKR)-like ER kinase (PERK), activating transcription factor 4 (ATF4) and TNF-receptor-associated factor 2 (TRAF2) as judged to I/R group. However, pre treatment with L-NAME abolished these beneficial effects of IPC against renal I/R insults. CONCLUSION: These findings suggest that early IPC protects kidney against renal I/R injury via reducing oxidative and ER stresses. These effects are associated with phosphorylation of Akt, eNOS activation and NO production contributing thus to HIF-1α stabilization. The beneficial impact of IPC was abolished when NO production is inhibited before IPC application.

Insulin like growth factor-1 increases fatty liver preservation in IGL-1 solution.

AIM: To investigate the benefits of insulin like growth factor-1 (IGF-1) supplementation to serum-free institut georges lopez-1 (IGL-1) solution to protect fatty liver against cold ischemia reperfusion injury. METHODS: Steatotic livers were preserved for 24 h in IGL-1 solution supplemented with or without IGF-1 and then perfused "ex vivo" for 2 h at 37degrees C. We examined the effects of IGF-1 on hepatic damage and function (transaminases, percentage of sulfobromophthalein clearance in bile and vascular resistance). We also studied other factors associated with the poor tolerance of fatty livers to cold ischemia reperfusion injury such as mitochondrial damage, oxidative stress, nitric oxide, tumor necrosis factor-α (TNF-α) and mitogen-activated protein kinases. RESULTS: Steatotic livers preserved in IGL-1 solution supplemented with IGF-1 showed lower transaminase levels, increased bile clearance and a reduction in vascular resistance when compared to those preserved in IGL-1 solution alone. These benefits are mediated by activation of AKT and constitutive endothelial nitric oxide synthase (eNOS), as well as the inhibition of inflammatory cytokines such as TNF-α. Mitochondrial damage and oxidative stress were also prevented. CONCLUSION: IGL-1 enrichment with IGF-1 increased fatty liver graft preservation through AKT and eNOS activation, and prevented TNF-α release during normothermic reperfusion.

Normal human gingival epithelial cells sense C. parapsilosis by toll-like receptors and module its pathogenesis through antimicrobial peptides and proinflammatory cytokines.

This study was designed to investigate the interaction between C. parapsilosis and human epithelial cells using monolayer cultures and an engineered human oral mucosa (EHOM). C. parapsilosis was able to adhere to gingival epithelial cells and to adopt the hyphal form in the presence of serum. Interestingly, when cultured onto the engineered human oral mucosa (EHOM), C. parapsilosis formed small biofilm and invaded the connective tissue. Following contact with C. parapsilosis, normal human gingival epithelial cells expressed high levels of Toll-like receptors (TLR)-2, -4, and -6, but not TLR-9 mRNA. The upregulation of TLRs was paralleled by an increase of IL-1beta, TNFalpha, and IFNgamma mRNA expression, suggesting the involvement of these cytokines in the defense against infection with C. parapsilosis. The active role of epithelial cells in the innate immunity against C. parapsilosis infection was enhanced by their capacity to express high levels of human beta-defensin-1, -2, and -3. The upregulation of proinflammatory cytokines and antimicrobial peptide expression may explain the growth inhibition of C. parapsilosis by the gingival epithelial cells. Overall results provide additional evidence of the involvement of epithelial cells in the innate immunity against C. parapsilosis infections.

Cytokine release and cytotoxicity in human keratinocytes induced by polycyclic aromatic hydrocarbons (1-methylpyrene and perylene).

The cytotoxic effects of two polycyclic aromatic hydrocarbons (PAH) (1-methyplyrene and perylene) were investigated on human skin keratinocytes. Normal human keratinocytes were cultured in the presence of various concentrations of 1-methylpyrene and perylene either alone or in combination. Following incubation, keratinocyte adhesion, viability, proliferation, colony-forming efficiency, and apoptosis/necrosis level were examined. The effects of PAH on wound healing were also determined in vitro using a scrape-wound healing assay on epidermis-like tissue. In addition, the inflammatory cell response to PAH insult was examined through interleukin-1 (IL-1) alpha and interleukin-6 (IL-6) secretion. Each individual PAH significantly decreased keratinocyte adhesion and viability in a concentration-dependent manner, which was associated with a reduced ability of keratinocytes to proliferate and form colonies. When PAH were combined, a greater effect on keratinocyte adhesion, viability, and proliferation was noted. Decreased cell proliferation/colony-forming efficiency was accompanied by increased cell apoptosis following incubation with either PAH. This effect was enhanced by the inhibitory influence on keratinocyte migration, as assessed by culture scratching. Each PAH also exerted a significant effect on keratinocyte immune functions by modulating the secretion of inflammatory mediators. Indeed, 1-methylpyrene or perylene, individually or when combined, significantly upregulated IL-1alpha and IL-6 secretion. This effect was greater and was concentration dependent when the PAH combination was used. Overall results indicate that 1-methylpyrene and perylene exerted a cytotoxic effect on human keratinocytes. Our findings may shed light on mechanisms underlying potential adverse effects of 1-methylpyrene and perylene on human skin.

Candida famata modulates toll-like receptor, beta-defensin, and proinflammatory cytokine expression by normal human epithelial cells.

Candida albicans is no longer the only yeast involved in infectious disorders, as others, such as C. famata, commonly associated with foods as well as terrestrial and marine environments, are being recognized as potential emerging pathogens that cause human candidiasis. We investigated the interaction between C. famata and human epithelial cells using monolayer cultures and an engineered human oral mucosa (EHOM). C. famata was able to adhere to gingival epithelial cells but failed to adopt the hyphal form in the presence/absence of proteins. Interestingly, when cultured onto the engineered human oral mucosa (EHOM), C. famata formed a biofilm and invaded the connective tissue. When normal human gingival epithelial cells were put in contact with C. famata, they expressed high levels of Toll-like receptors (TLR)-2, -4, and -6, but not TLR-9 mARN. The upregulation of TLRs was paralleled by an increase of IL-1beta and TNFalpha, but not IFNgamma mARN expression, suggesting the involvement of specific pro-inflammatory cytokines (IL-1beta and TNFalpha) in the defense against infection with C. famata. The active role of epithelial cells in the innate immunity against C. famata infection was enhanced by their capacity to express high levels of human beta-defensin (HBD)-1, -2, and -3. The upregulation of pro-inflammatory cytokines and antimicrobial peptide expression may explain the growth inhibition of C. famata by the gingival epithelial cells. Overall results provide additional evidence of the involvement of C. famata in the activation of innate immunity and the contribution of human epithelial cells in local defenses against such exogenous stimulations as C. famata infections.

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.

[Effect of ischaemic preconditioning and vitamin C on functional recovery of ischaemic kidneys]. / Effet du pré-conditionnement ischémique et de la vitamine C sur la reprise fonctionnelle des reins ischémiques.

OBJECTIVE: To present the results of ischaemic preconditioning (IPC), pharmacological preconditioning with vitamin C (Vit C) and a combination of the two modalities on functional recovery of rat kidneys after prolonged warm ischaemia. MATERIAL: Forty six rats were divided into 5 experimental groups. Kidneys of the sham group (n = 9) were only submitted to dissection of the pedicle. Ischaemia (60 min, n = 10) was induced by clamping both kidneys. IPC (n = 9) consisted of two successive cycles (5 min/5 min) of ischaemia/reperfusion (I/R). Vit C (100 mg/Kg, n = 9) was administered by intravenous injection 30 min before warm ischaemia. The Vit C + IPC (n= 9) group received the combination of both treatments. Tissue malonedialdehyde (MDA), plasma lactate dehydrogenase (LDH), tubular reabsorption of sodium (TRNa) and creatinine clearance (GFR) were evaluated after 120 min of kidney reperfusion. RESULTS: Ischaemic kidneys showed a significant increase of MDA and LDH concentrations and a significant reduction of GFR and TRNa compared to the sham group. The use of lPC or Fit C induced a significant improvement of renal function compared to the ischaemia group. The combination of Vit C + IPC induced a slight improvement of experimental parameters compared to those of the ischaemia group, but did not improve kidney functioning compared to the use of either IPC or Vit C alone. CONCLUSION: IPC and Fit C improve the functional parameters of ischaemic kidneys. However the protective effects are attenuated when the two treatments are combined.