UHPLC-HRMS-Based Untargeted Lipidomics Reveal Mechanism of Antifungal Activity of Carvacrol against Aspergillus flavus

Aspergillus flavus is a common contaminant in grain, oil and their products. Its metabolite aflatoxin B1 (AFB1) has been proved to be highly carcinogenic. Therefore, it is of great importance to find possible antifungal substances to inhibit the growth and toxin production of Aspergillus flavus. Carvacrol (CV) was reported as a potent antifungal monoterpene derived from plants. In this paper, the antifungal effects and mechanism of CV on Aspergillus flavus were investigated. CV was shown good inhibition on the growth of Aspergillus flavus and the production of AFB1. CV used in concentrations ranging from 0, 50, 100 and 200 μg/mL inhibited the germination of spores, mycelia growth and AFB1 production dose-dependently. To explore the antifungal mechanism of CV on Aspergillus flavus, we also detected the ergosterol content of Aspergillus flavus mycelia, employed Scanning Electron Microscopy (SEM) to observe mycelia morphology and utilized Ultra-High-Performance Liquid Chromatography-High-Resolution Mass Spectrometry (UHPLC-HRMS) to explore the lipidome profiles of Aspergillus flavus. The results showed that the production of ergosterol of mycelia was reduced as the CV treatment concentration increased. SEM photographs demonstrated a rough surface and a reduction in the thickness of hyphae in Aspergillus flavus treated with CV (200 µg/mL). In positive ion mode, 21 lipids of Aspergillus flavus mycelium were downregulated, and 11 lipids were upregulated after treatment with 200-µg/mL CV. In negative ion mode, nine lipids of Aspergillus flavus mycelium were downregulated, and seven lipids upregulated after treatment with 200-µg/mL CV. In addition, the analysis of different lipid metabolic pathways between the control and 200-µg/mL CV-treated groups demonstrated that glycerophospholipid metabolism was the most enriched pathway related to CV treatment.

Pre stimulation by gibberellic acid and the effect of extremely diluted agitated gibberellic acid on wheat stalk growth

Background: In previous multicentre studies[1,2], the influence of a homeopathic ultra high dilution of gibberellic acid on wheat growth was scrutinized. Data showed that this test dilution slowed down stalk growth when experiments were performed in autumn season. Aim: To test the hypothesis that pre treatment of grains with high concentrations of gibberellic acid will enhance the effect of the ultra high dilution of the plant hormone Methods: Grains of winter wheat (Triticum aestivum) were observed under the influence of extremely diluted gibberellic acid (10-30) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy (“G30x”). Analogously prepared water was used for control (“W30x”). Grains (500 or 1000 per group) were pre treated with (not agitated) gibberellic acid 10-5, 10-4 and 10-3 (“Ge-5, Ge-4, Ge-3”) or with water (“W”) for control prior to treatment with G30x or W30x. Seedlings were allowed to develop under standardized conditions for 7 days; plants were harvested and stalk lengths were measured. Results: With regard to pre treatment, it can be seen that the groups treated with Ge-3 showed most growth, followed by the Ge-4 groups, the Ge-5 groups and the non pre treated W groups. This decline is modulated by the application of G30x and W30x (figure 1). The lower the pre treatment concentration of G, the more marked is a slowing down effect of G30x versus W30x. Conclusion: The hypothesis that pre treatment of grains with high concentrations of gibberellic acid will enhance the effect of G30x had to be rejected. In contrast, G30x slowed down stalk growth in the W group with p < 0.001 but only moderately in the Ge-5 and Ge-4 group and not at all in the Ge-3 group. Keywords: homeopathy, wheat stalk growth, gibberellic acid, high dilution References: [1] Pfleger A., Hofäcker J., Scherer-Pongratz W., Lothaller H., Reich C., Endler P.C. The effect of extremely diluted agitated gibberellic acid (10e-30) on wheat seedling development – a two researcher study. Complementary Therapies in Medicine 2011; 19: 164-169. [2] Endler PC, Matzer W, Reich C, Reischl T, Hartmann AM, Thieves K, Pfleger A, Hofäcker J, Lothaller H, Scherer-Pongratz W. Seasonal variation of the effect of extremely diluted agitated gibberellic acid (10e-30) on wheat stalk growth – a multi researcher study. The Scientific World Journal 2011; 11: 1667-1678.

Intercomparison of Radiosensitization Induced by Gold and Iron Oxide Nanoparticles in Human Glioblastoma Cells Irradiated by 6 MV Photons

In this work, an intercomparison of sensitization effects produced by gold (GNP) and dextran-coated iron oxide (SPION-DX) nanoparticles in M059J and U87 human glioblastoma cells was performed using 6MV-photons. Three variables were mapped: the nanoparticle material, treatment concentration, and cell radiosensitivity. For U87, GNP treatments resulted in high sensitization enhancement ratios (SER10% up to 2.04). More modest effects were induced by SPION-DX, but still significant reductions in survival were achieved (maximum SER10%=1.61). For the radiosensitive M059J, sensitization by both NPs was poor. SER10% increased with the degree of elemental uptake in the cells, but not necessarily with treatment concentration. For GNP, where exposure concentration and elemental uptake were found to be proportional, SER10% increased linearly with concentration in both cell lines. For SPION-DX, saturation of sensitization enhancement and metal uptake occurred at high exposures. Fold change in the α/β ratios extracted from survival curves are reduced by the presence of SPION-DX but strongly increased by GNPs, suggesting that sensitization by GNPs occurs mainly via promotion of lethal damage, while for SPION-DX repairable damage dominates. The NPs were more effective in eliminating the radioresistant glioblastoma cells, an interesting finding, as resistant cells are key targets to improve treatment outcome.

Modification Mechanism and Rheological Properties of Emulsified Asphalt Evaporative Residues Reinforced by Coupling-Modified Fiber

In order to solve the problems of the smooth surface of basalt fiber and its weak interfacial adhesion with emulsified asphalt cold recycled mixture, a silane coupling agent (KH550) was used to treat the surface of basalt fiber and the effects of treatment concentration and soaking time on fiber modification were studied. The influence of silane coupling-modified basalt fiber (MBF) on the rheological properties of emulsified asphalt evaporation residue was studied at high and low temperatures using three routine index tests: a dynamic shear rheological test (DSR), a bending beam rheological test (BBR), and a force ductility test. The elemental changes of the fiber before and after modification and the microstructure of the emulsified asphalt evaporation residue with the coupling-modified fiber were analyzed by Fourier infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and X-ray energy dispersive spectroscopy (EDS), which is used to study the modification mechanism of emulsified asphalt evaporation residue reinforced by coupling-modified fiber. The results indicate that the concentration and soaking time of the silane coupling agent have a great influence on the surface morphology and mechanical properties of the fiber, and that the optimal treatment concentration is 1.0% and the optimal soaking time is 60 min. The addition of coupling-modified fibers can reduce the phase angle and unrecoverable creep compliance of emulsified asphalt evaporation residue, increase the rutting factor and creep recovery rate, and improve the elastic recovery ability and permanent deformation resistance. However, excessive fiber will weaken the ductility of emulsified asphalt at low temperatures. The appropriate content of silane coupling-modified fiber (MBF) is 1.5%. After silane coupling modification, the fiber surface becomes rough and cohesion is enhanced between the fiber and the emulsified asphalt base. Silane coupling-modified basalt fiber (MBF) acts as reinforcement for stability and bridging cracks.

Proteomic Analysis: Explosive Salt Accumulation in Leaves of Morus alba L. under Salt Stress

The salt tolerance of glycophytes is thought to be related to their ability to restrict sodium access to their aboveground parts. A previous study on the mulberry (Morus alba L.) revealed a phenomenon of explosive salt accumulation in the leaves after exceeding a certain treatment concentration. Here, we aim to observe the internal state of mulberry seedlings under salt stress by the proteomic method and to identify the possible inducements associated with salt bursts. In this study, the target treatments for TMT-label free quantitative analyses were determined by measuring the sodium content in the roots and leaves. The results showed that the expressions of proteins classified as “plant hormones”, “ion channels”, “REDOX homeostasis”, “cytoskeleton” and “cell wall” changed significantly after salt bursts. This phenotype is associated with the destruction of the apoplast, in which the assembly of the Casparian strip may be affected by the inhibition of some key proteins, indirectly increasing the rate of ion migration through the endodermis into the shoots.

Comparative Transcriptome Analysis of Two Contrasting Chinese Cabbage (Brassica rapa L.) Genotypes Reveals That Ion Homeostasis Is a Crucial Biological Pathway Involved in the Rapid Adaptive Response to Salt Stress

Salt is the most important limiting factor in plant yield and quality. Different Chinese cabbage cultivars appeared different salt tolerances, but there are few studies attempting to elucidate the mechanism underlying this phenomenon. In this study, 100 mmol L–1 NaCl was found to be the most suitable treatment concentration according to a sprouting bag test of 39 Chinese cabbage cultivars, and through comprehensive comparison and analysis, the relative values of fresh weight and electrolyte leakage in leaves proved to be convenient indicators for the identification of salt tolerance in Chinese cabbage. We analyzed the physiological responses of Qinghua45 (salt-tolerant) and Biyuchunhua (salt-sensitive) in terms of the growth indexes, ion homeostasis and Photosynthesis, the results indicated that Qinghua45 could ensure osmotic regulation, ion homeostasis and photosynthesis under salt stress. Next, we compared the transcriptome dynamics of the two cultivars. Overall, 2,859 differentially expressed genes (DEGs) were identified, and the number of DEGs in Qinghua45 was significantly less than that in Biyuchunhua. VDAC promoted the release of Ca2+, which indirectly promoted the transport of Na+ to vacuoles through the SOS2 pathway. Cation/H (+) antiporter 17 and V-H + -ATPase improve the exchange of Na+ and H+ and maintain Na+ in the vacuoles, thereby reducing the injury affected by salt stress. Increases in galactinol synthase and soluble protein synthesis helped relieve osmotic stress caused by salt, together, they regulated the Na+ content and chlorophyll biosynthesis of the plant and enabled the plant to adapt to salt stress over time.

Impact of Adjuvant Treatment on Heparanase Concentration in Invasive, Unilateral Breast Cancer Patients: Results of a Prospective Single-Centre Cohort Study

Background: In recent years, great progress has been made in the treatment of breast cancer, but it is still one of the ten leading causes of death in women. The aim of the study was to evaluate the heparanase concentration of invasive breast cancer (IBrC) patients, before and after cancer adjuvant treatment. Methods: Eighty patients with stage IA to IIB IBrC receiving adjuvant treatment were included prospectively in this study. The heparanase concentrations were determined by an enzyme-linked immunosorbent assay. A univariate analysis was used to estimate the factors influencing the low or high pre-treatment concentration of heparanase and the low or high numerical decrease in heparanase concentration after completion of adjuvant treatment. Results: Treatment reduced the concentration of heparanase by almost four times in the general IBrC cohort. Higher levels of pre- and post-treatment heparanase were noted in oestrogen receptor-negative cancers than in positive ones. A higher post-treatment concentration of heparanase was found in patients with a triple-negative tumour compared to patients with a luminal B HER2 negative type of IBrC. Overweight IBrC subjects and those with a tumour diameter of ≥2 cm demonstrated a lower chance of a lower pre-treatment heparanase concentration. Interestingly, a pre-treatment heparanase concentration is the main predictor of the changes in heparanase concentration after adjuvant treatment. Follow-up revealed significantly lower progression-free survival (PFS) rates in IBrC patients with a pre-treatment concentration of heparanase higher than 181.46 pg/mL (PFS = 80%). Conclusions: Our findings provide supporting evidence that IBrC therapy reduced the heparanase levels, regardless of treatment patterns and a pre-treatment concentration of heparanase may serve as a prognostic indicator for future outcomes.

EKSTRAK KARAGINAN RUMPUT LAUT (Eucheuma cottonii) YANG DIPEROLEH DARI PANTAI TABLOLONG

ABSTRACTEucheuma cottonii (E. cottonii) seaweed is a low level plant that has a high nutritional value. One of the ingredients that play a role in the formation of texture is carrageenan.Carrageenan is a type of hydrocolloid extracted from the seaweed of the red algae group (Rhodophyceae). The purpose of this study was to determine the quality of yields of Seaweed Carrageenan Extract (E. cottonii) obtained from Tablolong beach in the form of yield, ash content and fat content. The method used in this study is an experimental method with a completely randomized design (CRD) and for fat testing using quantitative descriptive methods. This study uses various concentrations of 0.1 N KOH, 0.2 N KOH, 0.3 N KOH and 0.4 N KOH. The results of the carrageenan yield test show that the addition of 0.1 N KOH concentration has a value of 56.10, KOH 0 , 2 N 62.16, KOH 0.3 N 72.73 and the best treatment was at P4 KOH 0.4 N 80.90. Carrageenan ash content test results showed that the addition of KOH concentration value obtained P1 KOH 0.1 N was 60.50, P2 KOH 0.2 N was 70.70, P3 KOH 0.3 N was 80.50, and P4 KOH 0.4 with a value of 90.50. The results of the carrageenan fat content of 0.1 KOH treatment has a fat content of 0, 2 while the treatment of KOH 0,2, KOH 0,3, KOH 0,4 have no fat content by showing the value (0). The results showed that the highest yield in the treatment of KOH concentration of 0.4 N with a value 80.95% and carrageenan ash content has the highest value in the treatment concentration of 0.4 N with a value of 90.50% while the fat content has a value of 0.2 in the treatment of KOH concentration of 0.1 N Keywords: Carrageenan, extract of E. cottonii seaweed.

PSVI-9 Effects of Mixing Prior to Automated Packaging in Extended Boar Semen

It is known that sperm cells will settle out of extender over time which could impact the concentration per dose within a batch. The objective of this study to investigate the effects of mixing prior to packaging on concentration of packaged insemination doses. At a commercial boar stud, three ejaculates were collected, extended, pooled and assigned to one of three treatment groups: pour-mix (PM) where ejaculates were immediately packaged after combining the three ejaculates, stir-mix (SM) where three ejaculates were combined and stirred thoroughly then packaged, or no mixing (NM) where pooled ejaculates were allowed to sit for 10 min then packaged. Semen was packaged with 35ml per tube creating ~350 individual doses of semen. Every fifth dose was labeled with consecutive numbers (~70/treatment) and overnighted to Purdue University. Concentration of each dose was determined using the NucleoCounter® SP-100TM. The entire study was repeated over 6 weeks. Within treatment, concentration did not differ among doses for the PM and SM treatments. However, NM doses were less concentrated (P&lt; 0.001) in doses &gt;300 compared to all others and doses 251–300 were less concentrated than doses 1 to 50. NM samples 1–250 averaged 36.12 million cells/ml whereas samples 251 to the end of the batch averaged 34.68 million/ml. This results in the last 20 doses having ~50.4 million less cells/dose. Across treatments, NM had lower concentrations (P=0.02) from doses &gt;200 compared to the SM treatment. PM appeared to be an intermediate as it was numerically lower than SM, but not statistically different from either NM or SM. The difference in concentrations between the NM and SM &gt;200 was 1.85 million/ml equaling 64.75 million fewer cells. Overall, allowing semen to settle and not mixing prior to packaging can result in 50–64 million cells fewer per dose in the last 50–100 doses packaged.

Surface Modification of Areca Fibre by Benzoyl Peroxide and Mechanical Behaviour of Areca-Epoxy Composites

Natural fibre composites are playing great role in current life scenario where the focus is more on replacing synthetic fibre composites with natural fibre composites. In this current study, investigation was done on tensile and flexural behaviour of benzoyl peroxide treated areca sheath fibre epoxy composites. The surface modification of the fibre was confirmed by FTIR analysis. Treatment concentration was the major criteria which effects mechanical properties of the composites. At 4% concentration tensile strength and flexural strength was found to be maximum which was reported as 37.05 N/mm2 and 235.5 N/mm2 respectively which gradually decreased with increasing concentration of benzoyl peroxide. SEM analysis proved that at lesser concentration, the bonding between fibre and resin was effective which reduced as the concentration of benzoyl peroxide increased. This results in ineffective stress transfer between reinforcing material and the matrix which was the reason for failure of composites manufactured at higher treatment concentration.