Autochthonous strains of Trichoderma isolated from tannery solid waste improve phytoextraction potential of heavy metals by sunflower.

This research work was aimed at isolating and demonstrating the significant potential of autochthonous fungi for phytoextraction of hazardous metals in metal polluted soil using Helianthus annuus. Four multi-metal resistant strains of Trichoderma were selected from a total of 21 strains isolated from tannery polluted soil and tannery solid waste. Autochthonous Trichoderma strains were used singly and in the form of consortium (TC). Sunflower was grown in pots for 90 days having eight different amendments of tannery polluted soil with and without Trichoderma inoculation. Growth and biochemical attributes of the plants were observed along with metal content extract by different plant parts. The results revealed that TC enhanced shoot length, shoot dry weight, and metal uptake as compared to single specie inoculation. Similarly, BCF (72.8-118.23%) and TF were significantly pronounced in shoots of H. annuus grown with TC at 40% amended soil. The biochemical analysis of the plants showed that Trichoderma strains boosted the enzymatic (catalase, peroxidase, and superoxide dismutase) antioxidants in the plants. The use of indigenous fungi with metal accumulating plants like sunflower can help to alleviate metal contamination from industrial sites and can make the soil cultivable for energy crops.

The genus Trichoderma is among the most common cosmopolitan soil fungi that enhance phytoextraction capability of plants. Hence, the isolation and identification of diversified and potent Trichoderma strains from contaminated environments is the need of the hour for broad spectrum applications in bioremediation. In the present study, contaminated soil mycoflora was explored and multi-metal resistant strains of Trichoderma were isolated. Their application in myco-assisted phytoextraction with Helianthus annuus was assessed to analyze their impact on the metal removal efficacy and enhancing growth in highly contaminated soil.

Equilibrium kinetics and thermodynamic studies on biosorption of heavy metals by metal-resistant strains of Trichoderma isolated from tannery solid waste.

This study was aimed at finding the metal sorption potential of six indigenous Trichoderma strains by using batch experiments for Cd (II), Cr (VI), Cu (II), and Pb (II). Trichoderma atrobrunneum showed maximum metal biosorption potential at 800 mg L-1 of initial concentration. Two adsorption isotherm models, (1) Langmuir (2) Freundlich models, were employed on the biosorption data obtained at various initial metal concentrations (10 mg L-1-200 mg L-1) and pseudo-first (PSI) and pseudo-second (PSII) order equilibrium kinetic models were subjected to data of agitation time (3-7 days). A maximum correlation coefficient value (R2) of ≤ 1 was observed for the Langmuir and PSII model. Results revealed that pH 6-7 was the best for metal sorption, while metal removal efficiency was increased by increasing temperature (298 K, 303 K, 308 K, 313 K). The results of thermodynamic study parameters (∆G°, ∆H°, ∆S°) indicated that heavy metal biosorption by Trichoderma strains was an endothermic, spontaneous, and feasible process. Moreover, surface characterization analysis through SEM, BET, FTIR, and XRD showed that T. atrobrunneum and Trichoderma sp. could adsorb more metal ions when grown in high metal concentrations. The results indicate that living biomass of T. atrobrunneum and Trichoderma sp. is an effective multi-metal biosorbent that can be used for efficacious bioremediation of bio-treatment of heavy metal polluted wastewater.

A comparison of total RNA extraction methods for RT-PCR based differential expression of genes from Trichoderma atrobrunneum.

The genus Trichoderma is ubiquitous in various niches and is currently used for biocontrol, biofertilizer, enzyme production and bioremediation. However, molecular mechanisms underlying its diverse biological functions are yet not fully elucidated. Extraction of high-quality RNA for downstream applications such as quantitative polymerase chain reaction is a prerequisite. The current study aims to optimize a total RNA extraction protocol for high-quality and quantity RNA from Trichoderma atrobrunneum. Seven RNA extraction protocols including Trizol, RiboEx PureLink RNA mini kit, high salt CTAB (cetyltrimethylammonium bromide), modified high salt CTAB, SDS (sodium dodecyl sulfate) and CTAB-PVP (polyvinylpyrrolidone) were performed separately, to extract RNA. Quality and quantity of extracted RNA samples were further analyzed by Nanodrop spectrophotometry, agarose gel electrophoresis and RT-PCR analysis. The results of quantitative and qualitative analysis of RNA samples showed that more intact, high-quality RNA was extracted using the modified high salt CTAB as compared to other methods. The RT-PCR results for the amplification of the genes encoding ß-tubulin and Ubiquitin carrier protein also showed lowest threshold cycle (Ct) and coefficient of variation (CV) for RNA samples extracted with the modified high salt CTAB method as compared to RNA samples extracted with other protocols. Therefore, it is proposed that the modified high salt CTAB protocol is an excellent method to obtain high-quality RNA with good yield from T. atrobrunneum for its downstream applications. Moreover, the optimized protocol is very economical and can be used to extract total RNA from a large number of samples.

miR-15a/-16 Inhibit Angiogenesis by Targeting the Tie2 Coding Sequence: Therapeutic Potential of a miR-15a/16 Decoy System in Limb Ischemia.

MicroRNA-15a (miR-15a) and miR-16, which are transcribed from the miR-15a/miR-16-1 cluster, inhibit post-ischemic angiogenesis. MicroRNA (miRNA) binding to mRNA coding sequences (CDSs) is a newly emerging mechanism of gene expression regulation. We aimed to (1) identify new mediators of the anti-angiogenic action of miR-15a and -16, (2) develop an adenovirus (Ad)-based miR-15a/16 decoy system carrying a luciferase reporter (Luc) to both sense and inhibit miR-15a/16 activity, and (3) investigate Ad.Luc-Decoy-15a/16 therapeutic potential in a mouse limb ischemia (LI) model. LI increased miR-15a and -16 expression in mouse muscular endothelial cells (ECs). The miRNAs also increased in cultured human umbilical vein ECs (HUVECs) exposed to serum starvation, but not hypoxia. Using bioinformatic tools and luciferase activity assays, we characterized miR-15a and -16 binding to Tie2 CDS. In HUVECs, miR-15a or -16 overexpression reduced Tie2 at the protein, but not the mRNA, level. Conversely, miR-15a or -16 inhibition improved angiogenesis in a Tie2-dependent manner. Local Ad.Luc-Decoy-15a/16 delivery increased Tie2 levels in ischemic skeletal muscle and improved post-LI angiogenesis and perfusion recovery, with reduced toe necrosis. Bioluminescent imaging (in vivo imaging system [IVIS]) provided evidence that the Ad.Luc-Decoy-15a/16 system responds to miR-15a/16 increases. In conclusion, we have provided novel mechanistic evidence of the therapeutic potential of local miR-15a/16 inhibition in LI.

N-Acetyl Cysteine Inhibits Endothelin-1-Induced ROS Dependent Cardiac Hypertrophy through Superoxide Dismutase Regulation.

OBJECTIVE: Oxidative stress down regulates antioxidant enzymes including superoxide dismutase (SOD) and contributes to the development of cardiac hypertrophy. N-Acetyl cysteine (NAC) can enhance the SOD activity, so the aim of this study is to highlight the inhibitory role of NAC against endothelin-1 (ET-1)-induced cardiac hypertrophy. MATERIALS AND METHODS: In this experimental study at QAU from January, 2013 to March, 2013. ET-1 (50 µg/kg) and NAC (50 mg/kg) were given intraperitoneally to 6-day old neonatal rats in combination or alone. All rats were sacrificed 15 days after the final injection. Histological analysis was carried out to observe the effects caused by both drugs. Reactive oxygen species (ROS) analysis and SOD assay were also carried out. Expression level of hyper- trophic marker, brain natriuretic peptide (BNP), was detected by western blotting. RESULTS: Our findings showed that ET-1-induced cardiac hypertrophy leading towards heart failure was due to the imbalance of different parameters including free radical-induced oxidative stress and antioxidative enzymes such as SOD. Furthermore NAC acted as an antioxidant and played inhibitory role against ROS-dependent hypertrophy via regulatory role of SOD as a result of oxidative response associated with hypertrophy. CONCLUSION: ET-1-induced hypertrophic response is associated with increased ROS production and decreased SOD level, while NAC plays a role against free radicals-induced oxidative stress via SOD regulation.

Stress-responsive factor regulation in patients suffering from type 2 diabetes and myocardial infarction.

BACKGROUND/AIM: Pro-free radical oxidative stresses, as well as regulatory factors, are believed to be the key players in the development of diabetes and heart-related disorders such as myocardial infarction. The aim of the present study was to highlight the role of oxidative stress-responsive factors (reactive oxygen species [ROS], super oxide dismutase [SOD], and calpain-1) in type 2 diabetes and myocardial infarction. MATERIALS AND METHODS: A total of 100 type 2 diabetes patients with myocardial infarction and 50 normal individuals were selected for this analysis. The levels of ROS and activities of SOD in the serum were determined. Serum calpain-1 expression was checked using western blotting. RESULTS: The serum level of ROS and the expression of calpain-1 were significantly higher while the activity of SOD was significantly lower in diabetic patients with myocardial infraction compared to normal individuals. CONCLUSION: These findings suggest a possible link between decreased antioxidant (SOD) and increased ROS levels as well as calpain-1 expression, supporting the role of oxidative stress-regulatory factors in diabetes and myocardial infraction.

Tea and its consumption: benefits and risks.

The recent convention of introducing phytochemicals to support the immune system or combat diseases is a centuries' old tradition. Nutritional support is an emerging advancement in the domain of diet-based therapies; tea and its constituents are one of the significant components of these strategies to maintain the health and reduce the risk of various malignancies. Tea is the most frequently consumed beverage worldwide, besides water. All the three most popular types of tea, green (unfermented), black (fully fermented), and oolong (semifermented), are manufactured from the leaves of the plant Camellia sinensis. Tea possesses significant antioxidative, anti-inflammatory, antimicrobial, anticarcinogenic, antihypertensive, neuroprotective, cholesterol-lowering, and thermogenic properties. Several research investigations, epidemiological studies, and meta-analyses suggest that tea and its bioactive polyphenolic constituents have numerous beneficial effects on health, including the prevention of many diseases, such as cancer, diabetes, arthritis, cardiovascular disease (CVD), stroke, genital warts, and obesity. Controversies regarding beneficialts and risks of tea consumption still exist but the limitless health-promoting benefits of tea outclass its few reported toxic effects. However, with significant rise in the scientific investigation of role of tea in human life, this review is intended to highlight the beneficial effects and risks associated with tea consumption.

Interplay of Phosphorylated Apoptosis Repressor with CARD, Casein Kinase-2 and Reactive Oxygen Species in Regulating Endothelin-1-Induced Cardiomyocyte Hypertrophy.

OBJECTIVE(S): The role of the Apoptosis repressor with caspase recruitment domain (ARC) in apoptosis and in certain hypertrophic responses has been previously investigated, but its regulation of Endothelin-1 induced cardiac hypertrophy remains unknown. The present study discusses the inhibitory role of ARC against endothelin-induced hypertrophy. RESULTS: In present study Endothelin treated cardiomyocytes were used as a hypertrophic model, that were subsequently treated with adenovirus ARC and its mutant at different multiplicity of infections. Casein-kinase-2 inhibitors were used to produce dephosphorylated ARC and to study its effect on hypertrophy. Hypertrophy was assessed by cell surface area measurement, Atrial-natriuretic-Factor mRNA analysis and total protein assay. Reactive oxygen species analysis was carried out using the dichlorofluorescin-diacetate (DCFH-DA) assay. Over expression of ARC significantly inhibits Endothelin-induced cardiomyocyte hypertrophy. The nonphosphorylated mutant ARC (T149 A) remained unable to control endothelin-induced hypertrophy, suggesting a vital role for ARC phosphorylation in regulation of its activity. Sensitization study has been carried out to check the role of endogenous ARC using casein-kinase inhibitors. Finally, the significant role of ARC in regulating reactive oxygen species -mediated control of endothelin induced hypertrophy has also been assessed. Conclusion : Conclusively, present study showed the vital and potential therapeutic interventional role of ARC in preventing endothelin-1-induced cardiomyocyte hypertrophy. The regulation of hypertrophic pathway by ARC relies on blunting the reactive oxygen species attack. This study further suggests a mediatory role of casein-kinase-2 in Endothelin-induced hypertrophy, mainly through its phosphorylation of ARC.