Improvement of the Biosynthesis of Resveratrol in Endophytic Fungus (Alternaria sp. MG1) by the Synergistic Effect of UV Light and Oligomeric Proanthocyanidins

Resveratrol, a natural polyphenol compound with multiple bioactivities, is widely used in food and pharmaceutical industry. Endophytic fungus Alternaria sp. MG1, as a native producer of resveratrol, shows increasing potential application. However, strategies for improvement of the biosynthesis of resveratrol in this species are still scarce. In this study, different elicitors were used to investigate their effect on the biosynthesis of resveratrol in MG1 and the induction mechanism. Ultrasound and sodium butyrate had no effect and slight inhibition on the resveratrol production and related gene expression, respectively. UV radiation and co-culture with Phomopsis sp. XP-8 significantly promoted the biosynthesis of resveratrol with the highest production (240.57μg/l) coming from UV 20min. Co-culture altered the profiles of secondary metabolites in MG1 by promoting and inhibiting the synthesis of stilbene and lignin compounds, respectively, and generating new flavonoids ((+/−)-taxifolin, naringin, and (+)-catechin). Oligomeric proanthocyanidins (OPC) also showed an obviously positive influence, leading to an increase in resveratrol production by 10 to 60%. Two calcium-dependent protein kinases (CDPK) were identified, of which CDPK1 was found to be an important regulatory factor of OPC induction. Synergistic treatment of UV 20min and 100μm OPC increased the production of resveratrol by 70.37% compared to control and finally reached 276.31μg/l.

Antioxidant Activity, Polyphenolic Profiles and Antibacterial Properties of Leaf Extract of Various Paulownia spp. Clones

Paulownia spp. are widely distributed ornamental trees with leaves abundant in secondary metabolites of high medicinal potential. Eighteen breeding clones of Paulownia spp. were tested in terms of their antioxidant activity and total polyphenolic contents. The 50% ethanolic extracts (2 g/30 mL) of leaves and petioles were compared in the screening step. Eight paulownia clones were selected for detailed analyses including HPTLC polyphenolic profile, verbascoside content and antibacterial activity against five bacteria species (S. aureus, B. cereus, E. coli, Y. enterocolitica, S. enterica). The species-specific differences in terms of antioxidant activity correlated with phenolic compounds were found mainly in the case of leaf blade extracts, the highest for P. tomentosa × P. fortunei and the lowest for P. elongata × P. fortunei clones. The P. tomentosa clones varied greatly in this regard. In the HPTLC polyphenolic profile, the occurrence of some polyphenols was proved and the specific verbascoside content was quantified (70 to 225 mg/g DW). The P. tomentosa × P. fortunei hybrids had the highest inhibitory activity, mainly against Gram-positive bacteria, whereas only slight inhibition of S. aureus growth was observed for P. elongata × P. fortunei clones. The obtained results indicate diverse suitability of paulownia clones as a source of active ingredients.

The Use of Waste Sorbents for Surface Wastewater Treatment to Intensify Plant Growth

The possibility of using sorbents from FOPS® filters, spent in the processes of purification of surface wastewater, is shown to intensify the growth of radish plants of the “French breakfast” variety. The ability of these waste sorbents, introduced into soil-ground substrates (SGS) as structure-forming additives enriched with microelements, was found to significantly stimulate the development of the root system with some slight inhibition of shoot growth. In the course of the experiments, the optimal value of the addition of sorbents spent in the processes of cleaning surface runoff (from roads) to the SGS was determined, which provides the greatest increase in the biomass of the root system of plants used as a test culture. It has been established that the accumulation of heavy metals (HMs) in the leaves and roots of a plant test culture increases with an increase in their content in the spent sorbents, but the actual concentrations of HMs in plants do not exceed the standards for their content in robust feed for animal husbandry. An assessment was made of the use of sorbents spent in water purification processes to create soil used in the reclamation of landfills for the placement of solid municipal waste, industrial dumps, road slopes, as well as in the restoration of forest resources and soil remediation.

DEMOGRAPHIC CHANGES IN THE RURAL AREAS OF THE WIELKOPOLSKIE PROVINCE IN THE YEARS 2005-2015

The aim of the research was to diagnose demographic changes that took place in the rural areas of the Wielkopolska province between 2005 and 2015. The study identified rural areas that were located within the boundaries of the Poznań Metropolitan Area and beyond this area. The research was based on data from the Central Statistical Office and the Statistical Office in Poznań. As a result of the conducted research, it was observed that demographic changes in the rural areas of Wielkopolska province such as population growth or changes in population structure by place of residence or economic age groups are largely caused by migration movements. In the analyzed period there was an influx of population into rural areas (positive migration balance), although in recent years a slight inhibition of this phenomenon has been observed. In addition, there was a slight increase in positive natural growth. More dynamic changes have been observed in rural areas within the Poznan Metropolitan Area. In the studied period, changes in the opposite direction were observed in cities, ie the decrease in the number of inhabitants and in birthrate, as well as the increase in emigration from cities (decrease of the negative migration balance).

Bioassay-Guided Chromatographic Isolation and Identification of Antibacterial Compounds from Artemisia annua L. That Inhibit Clostridium perfringens Growth

Clostridium perfringens is the causative agent of necrotic enteritis leading to significant losses in the poultry industry. Dichloromethane and n-hexane extracts of aerial parts of Artemisia annua (Asteraceae) exhibited activity against C. perfringens with minimum inhibitory concentrations (MIC) of 185 and 270 μg/mL, respectively. Bioassay-guided fractionation of the extracts gave several active fractions (MIC between 75 and 600 μg/mL). Investigations of the most active fractions resulted in the iso ponticaepoxide (MIC between 100 and 200 μg/mL) and (+)-threo-(5E)-trideca-1,5-dien-7,9,11-triyne-3,4-diol (MIC between 400 and 800 μg/mL), the flavonols chrysosplenol D (MIC between 200 and 400 μg/mL) and casticin (slight inhibition at 800 μg/mL), and 2,4-dihydroxy-6-methoxyacetophenone (slight inhibition at 800 μg/mL). Also, the coumarin scopoletin and the sesquiterpene lactone artemisinin were isolated from active fractions but showed no inhibition of C. perfringens growth at 800 and 2000 μg/mL, respectively. Fractions containing essential oil components with camphor constituting >60% did not show inhibition of C. perfringens up to 1600 μg/mL. Extracts and some active fractions showed higher antibacterial effect than individual bioactive compounds, suggesting that synergistic effects may underlie the observed antibacterial effect. The present study adds new valuable information on the antibacterial effect of A. annua against C. perfringens.

Combinatorial treatment using CTO with paclitaxel or paclitaxel orotate in sc-implanted human OVCAR-5 ovarian tumor model.

e15515 Background: Paclitaxel (PXL) was converted to paclitaxel orotate (PXLO). Different combinations of Carboxyamidotriazole orotate (CTO) and PXL or PXLO were first evaluated in female non-tumored athymic NCr-nu/nu mice to determine the tolerance of the combinations. The antiproliferative and antimetastatic effects of CTO are related to inhibition of receptor-operated calcium channel-mediated calcium influx. CTO can inhibit calcium sensitive signal transduction in the VEGF and the PI3K pathways, and inhibit FGF-2-induced tyrosine kinase. Methods: The antitumor effect of combinations was tested for: i) PXLO compared to PXL, ii) combinatorial effect of low (342mg/kg/dose) and high (513mg/kg/dose) doses of CTO (Q1Dx14, PO) with low (11.6 and 10mg/kg/inj, Q2Dx3/2 weeks, IV) and high (23.2 and 20mg/kg/inj, Q2Dx3/2 weeks, IV) respectively, doses of PXLO or PXL, against sc-implanted human OVCAR-5 ovarian tumor xenografts. Results: Results obtained in non-tumored mice show high PXLO plus CTO was better tolerated than high PXL plus CTO. In tumor bearing mice, oral CTO at doses of 513 or 342mg/kg/dose Q1Dx14 given alone resulted in slight inhibition of tumor growth, reaching statistical significance only when administered at a low dose (two tumor mass doubling: p=0.036 for 342mg/kg/dose). PXLO at 23.2 and 11.6mg/kg/inj. given alone inhibited tumor growth (p<0.001, 0.029, respectively). PXL at 20 and 10mg/kg/inj inhibited tumor growth (p<0.001 and 0.011, respectively). Since body weight loss was less with the orotate form of PXL, PXLO should be the preferred drug over PXL. Addition of oral CTO 513 or 342mg/kg/dose to PXLO 23.2 or PXL 20mg/kg/inj resulted in comparable tumor inhibition to PXLO or PXL alone. However, combination of PXLO 11.6 and PXL 10mg/kg/inj with CTO 513 or 342mg/kg/dose resulted in tumor inhibition suggesting that a low dose of PXL or PXLO may be combined with CTO to achieve tumor inhibition and to allow the use of the lower doses of PXL to reduce toxicity. Conclusions: Results suggest that PXLO is less toxic than PXL and CTO influences the sensitivity of PXL suggesting a role for combinatorial therapy with low doses of PXL and CTO in this ovarian tumor model.

Enhanced Hemostatic Performance of Tranexamic Acid-Loaded Chitosan/Alginate Composite Microparticles

Novel microparticles based on chitosan and sodium alginate were prepared using emulsification and cross-linking technologies. The spherical microparticles had a porous surface and a diameter of2~40 μm. In simulated body fluid, these microparticles quickly swelled but gradually degraded. The results of the MTT assay revealed that a slight inhibition of cell proliferation was observed on day 2 and then gradually decreased afterward. No cell morphology changes were observed. By loading tranexamic acid, the hemostatic performance of the microparticles was obviously improved. Using fast-acting styptic powder (Flashclot) as the control, the hemostatic efficiency was investigated in rabbits using a liver transection bleeding model. It was found that both Flashclot and the microparticles achieved hemostasis in3.07±0.84 min and2.48±0.88 min, respectively; however, the tranexamic acid-loaded microparticles stopped the bleeding in1.90±0.75 min (P<0.05). Additionally, Flashclot resulted in heat injury to the experimental livers, while the microparticles did not. Thus, with their biodegradability, safety, and superior hemostatic efficiency, tranexamic acid-loaded microparticles might be a promising new powdered hemostatic agent with a wide range of potential applications.

Novel Phenylazo Derivatives of Condensed and Uncondensed Thiophene. Synthesis, Characterization, and Antimicrobial Studies

In the search for new therapeutic agents against microbial infections, two novel series of monocyclic and tricyclic 5-(phenylazo)thiophene systems were synthesized based on 3-amino-2-thioxopyrimidinone and 2-cyanoacetamidothiophene derivatives 4 and 6. Functionalization of the pyrimidine ring in precursor 4 resulted in the formation of the target tricyclic condensed thiophenes 7, 12, and 13a, b, by the application of a variety of addition, substitution, and condensation reactions. On the other hand, derivatization of the versatile cyanoacetylated compound 6 led to a second series of monocyclic N-substituted aminothiophenes 15, 17, 19, and 20, through convenient methods. The new thiophene-based derivatives were tested for their antimicrobial activity with reference to relevant standard drugs. They displayed different levels of antibacterial activity, with compound 7 showing essentially the highest antipseudomonal activity. As for antifungal action, the compounds under investigation, unfortunately, had no inhibitory effects against test fungi isolates except for 7 and 20, that revealed slight inhibition.

3-Iodothyronamine metabolism and functional effects in FRTL5 thyroid cells

3-Iodothyronamine (T1AM), produced from thyroid hormones (TH) through decarboxylation and deiodination, is a potent agonist of trace amine-associated receptor 1 (TAAR1), a G protein-coupled receptor belonging to the family of TAARs.In vivoT1AM induces functional effects opposite to those produced on a longer time scale by TH and might represent a novel branch of TH signaling. In this study, we investigated the action of T1AM on thyroid and determined its uptake and catabolism using FRTL5 cells. The expression of TAAR1 was determined by PCR and western blot in FRTL5 cells, and cAMP, iodide uptake, and glucose uptake were measured after incubation with increasing concentrations of T1AM for different times. T1AM and its catabolites thyronamine (T0AM), 3-iodothyroacetic acid (TA1), and thyroacetic acid (TA0) were analyzed in FRTL5 cells by HPLC coupled to tandem mass spectrometry. The product of amplification ofTAAR1gene and TAAR1 protein was demonstrated in FRTL5 cells. No persistent and dose-dependent response to T1AM was observed after treatment with increasing doses of this substance for different times in terms of cAMP production and iodide uptake. A slight inhibition of glucose uptake was observed in the presence of 100 μM T1AM after 60 and 120 min (28 and 32% respectively), but the effect disappeared after 18 h. T1AM was taken up by FRTL5 cells and catabolized to T0AM, TA1, and TA0confirming the presence of deiodinase and amine oxidase activity in thyroid. In conclusion, T1AM determined a slight inhibition of glucose uptake in FRTL5 cells, but it was taken up and catabolized by these cells.

In vitro sugar uptake by grapefruit (Citrus paradisi) juice-sac cells

To further our understanding of the mechanisms of sugar uptake and accumulation into grapefruit (Citrus paradisi Macf. cv. Marsh seedless), the patterns of uptake and utilisation of sucrose, glucose and fructose by Citrus juice cells was investigated. Analyses were conducted on sliced juice sacs that were incubated in radioactive [14C]-sugar solutions with unlabelled sugars, in the presence or absence of metabolic inhibitors. Both hexoses demonstrated an initial uptake peak in December and a second uptake peak in February–March. From March through April the rates of sucrose uptake increased to levels comparable to those of glucose and fructose. Sucrose and its moieties fructose and glucose entered the juice cells of Citrus juice fruit by an insaturable, and mostly by an independent, process. However, NaN3 and carbonylcyanide m-chlorophenylhydrazone (CCCP) produced slight inhibition of these processes. Cells took up hexoses at a greater rate than sucrose, with accumulation reaching a plateau by 4–8 h, and then continuing unabated, in the case of glucose, for 42 h. Uptake of all three sugars increased linearly in the range of sugar concentrations tested, which extended from 0.01 to 320 mm, denoting an insaturable system for sugar uptake. 14CO2 evolution was relatively low in all the experiments, the lowest evolution being recorded when the uptake of [14C]-sucrose was studied, while the highest 14CO2 evolution was recorded when the uptake of [14C]-glucose was studied. The data demonstrate a preferential utilisation of glucose over fructose and sucrose. In all the experiments, the two metabolic inhibitors significantly inhibited the decarboxylation of the three sugars.