Nourin-Associated miRNAs: Novel Inflammatory Monitoring Markers for Cyclocreatine Phosphate Therapy in Heart Failure

Background: Cyclocreatine phosphate (CCrP) is a potent bioenergetic cardioprotective compound known to preserve high levels of cellular adenosine triphosphate during ischemia. Using the standard Isoproterenol (ISO) rat model of heart failure (HF), we recently demonstrated that the administration of CCrP prevented the development of HF by markedly reducing cardiac remodeling (fibrosis and collagen deposition) and maintaining normal ejection fraction and heart weight, as well as physical activity. The novel inflammatory mediator, Nourin is a 3-KDa formyl peptide rapidly released by ischemic myocardium and is associated with post-ischemic cardiac inflammation. We reported that the Nourin-associated miR-137 (marker of cell damage) and miR-106b-5p (marker of inflammation) are significantly upregulated in unstable angina patients and patients with acute myocardial infarction, but not in healthy subjects. Objectives: To test the hypothesis that Nourin-associated miR-137 and miR-106b-5p are upregulated in ISO-induced “HF rats” and that the administration of CCrP prevents myocardial injury (MI) and reduces Nourin gene expression in “non-HF rats”. Methods: 25 male Wistar rats (180–220 g) were used: ISO/saline (n = 6), ISO/CCrP (0.8 g/kg/day) (n = 5), control/saline (n = 5), and control/CCrP (0.8 g/kg/day) (n = 4). In a limited study, CCrP at a lower dose of 0.4 g/kg/day (n = 3) and a higher dose of 1.2 g/kg/day (n = 2) were also tested. The Rats were injected SC with ISO for two consecutive days at doses of 85 and 170 mg/kg/day, respectively, then allowed to survive for an additional two weeks. CCrP and saline were injected IP (1 mL) 24 h and 1 h before first ISO administration, then daily for two weeks. Serum CK-MB (U/L) was measured 24 h after the second ISO injection to confirm myocardial injury. After 14 days, gene expression levels of miR-137 and miR-106b-5p were measured in serum samples using quantitative real-time PCR (qPCR). Results: While high levels of CK-MB were detected after 24 h in the ISO/saline rats indicative of MI, the ISO/CCrP rats showed normal CK-MB levels, supporting prevention of MI by CCrP. After 14 days, gene expression profiles showed significant upregulation of miR-137 and miR-106b-5p by 8.6-fold and 8.7-fold increase, respectively, in the ISO/saline rats, “HF rats,” compared to the control/saline group. On the contrary, CCrP treatment at 0.8 g/kg/day markedly reduced gene expression of miR-137 by 75% and of miR-106b-5p by 44% in the ISO/CCrP rats, “non-HF rats,” compared to the ISO/Saline rats, “HF rats.” Additionally, healthy rats treated with CCrP for 14 days showed no toxicity in heart, liver, and renal function. Conclusions: Results suggest a role of Nourin-associated miR-137 and miR-106b-5p in the pathogenesis of HF and that CCrP treatment prevented ischemic injury in “non-HF rats” and significantly reduced Nourin gene expression levels in a dose–response manner. The Nourin gene-based mRNAs may, therefore, potentially be used as monitoring markers of drug therapy response in HF, and CCrP—as a novel preventive therapy of HF due to ischemia.     

Mechanisms of the Procognitive Effects of Xanthotoxin and Umbelliferone on LPS-Induced Amnesia in Mice

Neuroinflammation plays an essential role in the pathogenesis of neurodegenerative diseases such as Alzheimer’s disease. Although coumarins have been shown to improve cognitive function in animal models and exert anti-inflammatory effects in cell cultures, the exact mechanism of their neuroprotective effects has not yet been fully elucidated. The present study aimed to investigate the neuroprotective effects of xanthotoxin (furanocoumarin) and umbelliferone (simple coumarin) in lipopolysaccharide-induced cognitive dysfunction in mice. For evaluation memory and learning processes, a passive avoidance test was used. Furthermore, acetylcholinesterase level and impact on the tumor necrosis factor α, interleukin 10 levels in the whole brain, and cyclooxygenase-II in hippocampus was established. Subchronic administration of both coumarins (15 mg/kg) enhanced the learning and memory function, but only the xanthotoxin improved cognitive processes impaired by lipopolysaccharide (0.8 mg/kg) administration. Behavioral results stay in line with acetylcholinesterase level in the brain. A statistically significant decrease in the level of tumor necrosis factor α and cyclooxygenase-II in lipopolysaccharide-treated rodents after coumarins’ administration was observed. Together, our findings demonstrate that both coumarins improved cognitive functions, but only xanthotoxin significantly enhanced the learning and memory function and reduced the level of acetylcholinesterase in lipopolysaccharide-treated mice. This effect may suggest that only furanocoumarin—xanthotoxin attenuates neuroinflammation and enhances cholinergic neurotransmission, thus it can be a potential remedy with procognitive potential effective in treatment of neuroinflammatory disease.     

Fermentative hydrogen production by a new alkaliphilic Clostridium sp. (strain PROH2) isolated from a shallow submarine hydrothermal chimney in Prony Bay,New Caledonia

The hydrogen-producing strain PROH2 pertaining to the genus Clostridium was successfully isolated from a shallow submarine hydrothermal chimney (Prony Bay, New Caledonia) driven by serpentinization processes. Cell biomass and hydrogen production performances during fermentation by strain PROH2 were studied in a series of batch experiments under various conditions of pH, temperature, NaCl and glucose concentrations. The highest hydrogen yield, 2.71 mol H₂/mol glucose, was observed at initial pH 9.5, 37 °C, and glucose concentration 2 g/L, and was comparable to that reported for neutrophilic clostridial species. Hydrogen production by strain PROH2 reached the maximum production rate (0.55 mM-H₂/h) at the late exponential phase. Yeast extract was required for growth of strain PROH2 and improved significantly its hydrogen production performances. The isolate could utilize various energy sources including cellobiose, galactose, glucose, maltose, sucrose and trehalose to produce hydrogen. The pattern of end-products of metabolism was also affected by the type of energy sources and culture conditions used. These results indicate that Clostridium sp. strain PROH2 is a good candidate for producing hydrogen under alkaline and mesothermic conditions.     

Insecticidal activities of essential oils from leaves of Laurus nobilis L. from Tunisia,Algeria and Morocco,and comparative chemical composition

Laurus nobilis essential oils from Tunisia, Algeria and Morocco were analyzed for their chemical composition and assessed for their repellent and toxic activities against two major stored product pests: Rhyzopertha dominica and Tribolium castaneum. The three oils showed quantitative rather than qualitative differences in their chemical compositions. 1,8-cineole, linalool and isovaleraldehyde, were identified as the major common compounds whereas, α-pinene, α-terpineol, eugenylmethylether, β-pinene, spathulenol and β-myrcene were also well represented in all three oils. Results showed that L. nobilis essential oils were repellant and toxic to adults of R. dominica and T. castaneum. Repellent and fumigant toxicities were highly dependent upon insect species and oil origin.In filter paper tests, L. nobilis essential oil from Morocco was more effective compared to Tunisian and Algerian oils. RD₅₀ values were respectively 0.013 μl/cm², 0.036 μl/cm² and 0.033 μl/cm² for R. dominica versus 0.045 μl/cm², 0.139 μl/cm² and 0.096 μl/cm² for T. castaneum. Moreover, fumigant activity tests showed that both R. dominica and T. castaneum were more susceptible to L. nobilis essential oil from Morocco than that from Algeria or Tunisia. The corresponding LC₅₀ values were respectively 68, 99 and 113 μl/l air for R. dominica against 172, 194 and 217 μl/l air for T. castaneum. Our work clearly vindicates interest in the efficacy of essential oils from plants of Mediterranean origin both as insecticides and repellents against stored product pests.     

Chemical and aroma volatile compositions of date palm (Phoenix dactylifera L.) fruits at three maturation stages

The effect of maturity stage on the physicochemical composition and volatile components of date palm fruits (Phoenix dactylifera L.) was investigated at three different stages. Total sugars, pH, proteins and ash increased up to the full-ripe stage of all date types. On the contrary, weight, moisture and total lipids decreased across the same period. Pearson’s test established significant differences (p < 0.05) between the different cultivars at each stage for all the parameters. Furthermore, multivariate tests show that the composition and the variation of the chemical compositions mainly depended on the cultivar kind. Eighty volatile compounds were identified during the maturation stages, 43 of them were newly identified in Tunisian date fruits. The profiles of volatile compounds seem to be similar for Degla and Horra varieties. Results from this work revealed essential information about the richness of littoral cultivars in many nutrients and the possibility of their uses as a functional food ingredient.     

Effect of Air-Drying Temperature on Kinetics of Quality Attributes of Lemon (Citrus limon cv. lunari) Peels

The aim of this work was to determine the effect of air drying temperature (40 °C, 50 °C, and 60 °C) on the degradation kinetics of quality parameters of lemon peels. Drying at 40 °C was in favor of maximal water and oil retention capacities and lowest color loss (35%), but it also induced lower phenolic and flavonoids residual contents. First-order reaction model showed a good fit for lightness and yellowness color parameter. Zero-order reaction model fitted well redness color parameter and total color difference. A higher value of activation energy indicated greater heat sensitivity of lightness and redness during air drying.     

Lead removal from aqueous solutions by a Tunisian smectitic clay

The adsorption of Pb(2+) ions onto Tunisian smectite-rich clay in aqueous solution was studied in a batch system. Four samples of clay (AYD, AYDh, AYDs, AYDc) were used. The raw AYD clay was sampled in the Coniacian-Early Campanian of Jebel A?doudi in El Hamma area (South of Tunisia). AYDh and AYDs corresponds to AYD activated by 2.5 mol/l hydrochloric acid and 2.5 mol/l sulphuric acid, respectively. AYDc corresponds to AYD calcined at different temperatures (100, 200, 300, 400, 500 and 600 degrees C). The raw AYD clay was characterized by X-ray diffraction, chemical analysis, infrared spectroscopy and coupled DTA-TGA. Specific surface area of all the clay samples was determined from nitrogen adsorption isotherms. Preliminary adsorption tests showed that sulphuric acid and hydrochloric acid activation of raw AYD clay enhanced its adsorption capacity for Pb(2+) ions. However, the uptake of Pb(2+) by AYDs was very high compared to that by AYDh. This fact was attributed to the greater solubility of clay minerals in sulphuric acid compared to hydrochloric acid. Thermic activation of AYD clay reduced the Pb(2+) uptake as soon as calcination temperature reaches 200 degrees C. All these preliminary results were well correlated to the variation of the specific surface area of the clay samples. The ability of AYDs sample to remove Pb(2+) from aqueous solutions has been studied at different operating conditions: contact time, adsorbent amount, metal ion concentration and pH. Kinetic experiments showed that the sorption of lead ions on AYDs was very fast and the equilibrium was practically reached after only 20 min. The results revealed also that the adsorption of lead increases with an increase in the solution pH from 1 to 4.5 and then decreases, slightly between pH 4.5 and 6, and rapidly at pH 6.5 due to the precipitation of some Pb(2+) ions. The equilibrium data were analysed using Langmuir isotherm model. The maximum adsorption capacity (Q(0)) increased from 25 to 25.44 mg/g with increasing temperature from 25 to 40 degrees C. Comparative study between sulphuric acid activated clay (AYDs) and powder activated carbon (PAC) for the adsorption of lead was also conducted. The results showed that sulphuric acid activated clay is more efficient than PAC.     

Monitoring of genetically modified food and feed in the Tunisian market using qualitative and quantitative real-time PCR

Genetically modified organisms (GMO) invade more and more the agricultural production in the world. Although there are no legislations on GM labeling and cultivation of GM crops in Tunisia, the present study aims to check the status of GMO in Tunisian market using qualitative and quantitative real time-PCR (QRT-PCR). Three-hundred-sixty five samples were collected and different DNA extraction methods were adapted and optimized. Specific primers targeting 35S promoter from Cauliflower mosaic virus (CaMV) and nopaline synthase terminator from Agrobacterium tumefaciens (At) were used for the detection of the GMO insert and Taxon specific primers for the detection of plant species. Validated Taqman® probes (EU-RL) targeting event specific regions of the maize events MON810, Bt11, and the soybean event RRS were used for the quantification studies. Seven food and feed products showed different amounts of RRS (1.9%), MON810 (2.1%), and Bt11 (1.6%). The results demonstrate for the first time the presence of GMO in Tunisian markets reinforcing the need for the development of accurate quantitative methods in routine analyses.