Decellularized Wharton's jelly extracellular matrix as a promising scaffold for promoting hepatic differentiation of human induced pluripotent stem cells

Liver tissue engineering as a therapeutic option for restoring of damaged liver function has a special focus on using native decellularized liver matrix, but there are limitations such as the shortage of liver donor. Therefore, an appropriate alternative scaffold is needed to circumvent the donor shortage. This study was designed to evaluate hepatic differentiation of human induced pluripotent stem cells (hiPSCs) in decellularized Wharton's jelly (WJ) matrix as an alternative for native liver matrix. WJ matrices were treated with a series of detergents for decellularization. Then hiPSCs were seeded into decellularized WJ scaffold (DWJS) for hepatic differentiation by a defined induction protocol. The DNA quantitative assay and histological evaluation showed that cellular and nuclear materials were efficiently removed and the composition of extracellular matrix was maintained. In DWJS, hiPSCs-derived hepatocyte-like cells (hiPSCs-Heps) efficiently entered into the differentiation phase (G1) and gradually took a polygonal shape, a typical shape of hepatocytes. The expression of hepatic-associated genes (albumin, TAT, Cytokeratin19, and Cyp7A1), albumin and urea secretion in hiPSCs-Heps cultured into DWJS was significantly higher than those cultured in the culture plates (2D). Altogether, our results suggest that DWJS could provide a proper microenvironment that efficiently promotes hepatic differentiation of hiPSCs.     

Antagonistic activity and hyphal interactions of Trichoderma spp. against Fusarium proliferatum and F. oxysporum in vitro

Trichoderma is a well-known antagonist against soilborne plant pathogens. However, the species and even various isolates have different biocontrol potential. To evaluate the antagonistic activities of Trichoderma harzianum, T. harzianum strain T100 (T100), T. viride and T. haematum against Fusarium oxysporum and F. proliferatum, we used dual culture and productions of volatile and non-volatile metabolites in three different phases in vitro. An analysis of the data in dual culture tests represented T. viride, T. haematum and T100 as effective antagonists of Fusarium while T100 was the only fungus being able to lyse the confronting mycelia. Similar results were obtained in the volatile metabolites tests also. In contrast with the two previous tests, the non-volatile metabolites produced by T. harzianum inhibited Fusarium mycelial growth the most, and T100 acted moderately. It was also clearly showed that the antagonistic effect of Trichoderma spp. was more on F. proliferatum than on F. oxysporum. Finally, because Trichoderma spp. was most effective in the second phase, we recommend to use T100 against F. proliferatum at the initial stages of infection as its mycoparasitism on F. oxysporum was observed microscopically through forming apressoria structures without any coiling around the pathogen.     

Molecular Dynamics Study of the Human Beta-defensins 2 and 3 Chimeric Peptides with the Cell Membrane Model of Pseudomonas aeruginosa

International Journal of Peptide Research and Therapeutics - Pseudomonas aeruginosa is a unique microorganism among the antibiotic-resistant microbial pathogens. Among the various therapeutic...     

Sensitive microanalysis of gabapentin by high-performance liquid chromatography in human serum using pre-column derivatization with 4-chloro-7-nitrobenzofurazan: Application to a bioequivalence study

Most of the published methods for analysis of gabapentin, an antiepileptic agent, in human serum require automated o-phthalaldehyde derivatization of the drug and immediate injection of the unstable derivatives formed. A new, very sensitive and simple high-performance liquid chromatographic method for quantitation of the drug in human serum using 4-chloro-7-nitrobenzofurazan (NBD-Cl) as a fluorescent labeling agent is presented. In this method the sensitivity was significantly improved and the limit of quantification of 0.002 microg/ml was obtained using 100 microl serum sample and 10 microl injection. However, the LOQ can be improved by increasing the sampling volume. The procedure involved protein precipitation of serum by acetonitrile followed by derivatization with NBD-Cl. Amlodipine was used as internal standard and chromatographic separation was performed on a Shimpack CLC-C18 (150 mm x 4.6 mm) column. The fluorescence derivative of the drug was monitored at excitation and emission wavelengths of 470 and 537 nm, respectively. A mobile phase consisting of methanol and sodium phosphate buffer (0.05 M; pH 2.5) containing 1 ml/l triethylamine (65:35, v/v) was used. The calibration curve was linear over the concentration range of 0.002-15 microg/ml. No interferences were found from commonly co-administrated antiepileptic drugs. The method was applied in a randomized cross-over bioequivalence study of two different gabapentin preparations in 24 healthy volunteers.     

Chronic hyperammonemia reduces the activity of neuronal nitric oxide synthase in cerebellum by altering its localization and increasing its phosphorylation by calcium-calmodulin kinase II

Impaired function of the glutamate-nitric oxide-cGMP pathway contributes to cognitive impairment in hyperammonemia and hepatic encephalopathy. The mechanisms by which hyperammonemia impairs this pathway remain unclear. Understanding these mechanisms would allow designing clinical treatments for cognitive deficits in hepatic encephalopathy. The aims of this work were: (i) to assess whether chronic hyperammonemia in vivo alters basal activity of neuronal nitric oxide synthase (nNOS) in cerebellum and/or its activation in response to NMDA receptor activation and (ii) to analyse the molecular mechanisms by which hyperammonemia induces these alterations. It is shown that hyperammonemia reduces both basal activity of nNOS and its activation following NMDA receptor activation. Reduced basal activity is because of increased phosphorylation in Ser847 (by 69%) which reduces basal activity of nNOS by about 40%. Increased phosphorylation of nNOS in Ser847 is because of increased activity of calcium-calmodulin-dependent protein kinases (CaMKII) which in turn is because of increased phosphorylation at Thr286. Inhibiting CaMKII with KN-62 normalizes phosphorylation of Ser847 and basal NOS activity in hyperammonemic rats, returning to values similar to controls. Reduced activation of nNOS in response to NMDA receptor activation in hyperammonemia is because of altered subcellular localization of nNOS, with reduced amount in post-synaptic membranes and increased amount in the cytosol.     

Using aerated compost tea in comparison with a chemical pesticide for controlling rose powdery mildew

Rose powdery mildew (Sphaerotheca pannosa var. rosae) is one of the most common foliar diseases of roses worldwide. Application of chemical products on the plant or in the soil kills a range of the beneficial micro-organisms thereby disturbing ecosystem. Compost tea helps to restore and increase the populations of those beneficial micro-organisms. The objective of the present study was to evaluate the comparison of biopesticide (compost tea) and a chemical pesticide. The experiment was performed in three treatments, which were compost tea, fungicide (Topaz) and no treatment in three replications. After foliar applications of biopesticide and fungicide, the control percentage was estimated based on the number of infected flowers with powdery mildew. The results indicated that there was a significant difference between these treatments on rose in controlling powdery mildew (F?=?23.25, p?=?0.0015, df?=?2), at a probability level of 1% (p???0.01). So, that control percentage of compost tea treatment was the most.     

Phytosiderophore release by wheat genotypes differing in zinc deficiency tolerance grown with Zn-free nutrient solution as affected by salinity

There is limited information concerning the effect of salinity on phytosiderophores exudation from wheat roots. The aim of this hydroponic experiment was to investigate the effect of salinity on phytosiderophore release by roots of three bread wheat genotypes differing in Zn efficiency (Triticum aestivum L. cvs. Rushan, Kavir, and Cross) under Zn deficiency conditions. Wheat seedlings were transferred to Zn-free nutrient solutions and exposed to three salinity levels (0, 60, and 120 mM NaCl). The results indicated that Cross and Rushan genotypes exuded more phytosiderophore than did the Kavir genotype. Our findings suggest that the adaptive capacity of Zn-efficient ‘Cross’ and ‘Rushan’ wheat genotypes to Zn deficiency is due partly to the higher amounts of phytosiderophore release. Only 15 days of Zn deficiency stress was sufficient to distinguish between Zn-efficient (Rushan and Cross) and Zn-inefficient (Kavir) genotypes, with the former genotypes exuding more phytosiderophore than the latter. Higher phytosiderophore exudation under Zn deficiency conditions was accompanied by greater Fe transport from root to shoot. The maximum amount of phytosiderophore was exuded at the third week in ‘Cross’ and at the fourth week in ‘Kavir’ and ‘Rushan’. For all three wheat genotypes, salinity stress resulted in higher amounts of phytosiderophore exuded by the roots. In general, for ‘Kavir’, the largest amount of phytosiderophore was exuded from the roots at the highest salinity level (120 mM NaCl), while for ‘Cross’ and ‘Rushan’, no significant difference was found in phytosiderophore exudation between the 60 and 120 mM NaCl treatments. More investigation is needed to fully understand the physiology of elevated phytosiderophore release by Zn-deficient wheat plants under salinity conditions.     

Behavioral deficits induced by lead exposure are accompanied by serotonergic and cholinergic alterations in the prefrontal cortex

The effects of long-term lead (Pb) exposure producing a blood Pb concentration of lower than 20 μg/dL, i.e. below that associated with overt neurological deficits in occupationally exposed individuals, was studied in adult rats. In order to assess gender differences, we performed parallel behavioral experiments in male and female rats. Exposure to Pb acetate (50 ppm in drinking water) for 6 months induced motor and cognitive alterations, however these effects were gender- and task-dependent. Chronic lead exposure impaired spatial learning assessed in the Morris water maze test (MWM) in both genders, whereas it only induced hyperactivity in the open field and impaired motor coordination in the rotarod test, only in male rats. Hyperactivity in male rats was accompanied by an increase in extracellular level of acetylcholine in the prefrontal cortex. Extracellular dopamine concentration in the prefrontal cortex was unaffected by lead exposure whereas serotonin concentration in the same brain area was significantly decreased in both male and female rats exposed to lead. These results unveil new molecular mechanisms underlying neuropsychiatric alterations induced by chronic lead exposure.