A New HPLC-MS Method for Measuring Maslinic Acid and Oleanolic Acid in HT29 and HepG2 Human Cancer Cells

Maslinic acid (MA) and oleanolic acid (OA), the main triterpenic acids present in olive, have important properties for health and disease prevention. MA selectively inhibits cell proliferation of the HT29 human colon-cancer cell line by inducing selective apoptosis. For measuring the MA and OA concentration inside the cell and in the culture medium, a new HPLC-MS procedure has been developed. With this method, a determination of the amount of MA and OA incorporated into HT29 and HepG2 human cancer-cell lines incubated with different concentrations of MA corresponding to 50% growth inhibitory concentration (IC₅₀), IC_50/2, IC_50/4, and IC_50/8 has been made. The results demonstrate that this method is appropriate for determining the MA and OA concentration in different types of cultured cells and reveals the specific dynamics of entry of MA into HT29 and HepG2 cells.     

Trans-Splicing Improvement by the Combined Application of Antisense Strategies

Spliceosome-mediated RNA trans-splicing has become an emergent tool for the repair of mutated pre-mRNAs in the treatment of genetic diseases. RNA trans-splicing molecules (RTMs) are designed to induce a specific trans-splicing reaction via a binding domain for a respective target pre-mRNA region. A previously established reporter-based screening system allows us to analyze the impact of various factors on the RTM trans-splicing efficiency in vitro. Using this system, we are further able to investigate the potential of antisense RNAs (AS RNAs), presuming to improve the trans-splicing efficiency of a selected RTM, specific for intron 102 of COL7A1. Mutations in the COL7A1 gene underlie the dystrophic subtype of the skin blistering disease epidermolysis bullosa (DEB). We have shown that co-transfections of the RTM and a selected AS RNA, interfering with competitive splicing elements on a COL7A1-minigene (COL7A1-MG), lead to a significant increase of the RNA trans-splicing efficiency. Thereby, accurate trans-splicing between the RTM and the COL7A1-MG is represented by the restoration of full-length green fluorescent protein GFP on mRNA and protein level. This mechanism can be crucial for the improvement of an RTM-mediated correction, especially in cases where a high trans-splicing efficiency is required.     

Incidence of an Ozonation Stage on the Treatment of Cherry Stillage by Activated Sludge

This article presents a laboratory study of the ozonation of diluted cherry stillage, a high-strength wastewater. Influence of variables, kinetics, and the effects of an ozonation stage coupled with the biological treatment by activated sludge are addressed. Single activated sludge processing was shown effective to remove biological oxygen demand (BOD) and chemical oxygen demand (COD) but polyphenols were reduced to a lesser extent. On the other hand, direct wastewater ozonation did not reduce COD and total organic carbon (TOC) appreciably, and foaming problems were experienced when a high gas flow rate was applied. However, polyphenols and UV₂₅₄ absorbance decreased substantially by means of ozonation. To best achieve complete cherry stillage purification, two ways of coupling ozonation with activated sludge are proposed. Ozonation prior to activated sludge is advised for high-concentration wastewater to reduce polyphenol concentration, thus removing inhibiting effects. For wastewater with low polyphenol concentration the sequence activated sludge–ozonation–activated sludge is preferred to enhance the overall process performance in terms of oxidation efficiency and sludge settling.     

Pharmacogenetics Biomarkers and Their Specific Role in Neoadjuvant Chemoradiotherapy Treatments: An Exploratory Study on Rectal Cancer Patients

Background: Pathological complete response (pCR) to neoadjuvant chemoradiotherapy (CRT) in locally advanced rectal cancer (LARC) is still ascribed to a minority of patients. A pathway based-approach could highlight the predictive role of germline single nucleotide polymorphisms (SNPs). The primary aim of this study was to define new predictive biomarkers considering treatment specificities. Secondary aim was to determine new potential predictive biomarkers independent from radiotherapy (RT) dosage and cotreatment with oxaliplatin. Methods: Thirty germ-line SNPs in twenty-one genes were selected according to a pathway-based approach. Genetic analyses were performed on 280 LARC patients who underwent fluoropyrimidine-based CRT. The potential predictive role of these SNPs in determining pathological tumor response was tested in Group 1 (94 patients undergoing also oxaliplatin), Group 2 (73 patients treated with high RT dosage), Group 3 (113 patients treated with standard RT dosage), and in the pooled population (280 patients). Results: Nine new predictive biomarkers were identified in the three groups. The most promising one was rs3136228-MSH6 (p = 0.004) arising from Group 3. In the pooled population, rs1801133-MTHFR showed only a trend (p = 0.073). Conclusion: This exploratory study highlighted new potential predictive biomarkers of neoadjuvant CRT and underlined the importance to strictly define treatment peculiarities in pharmacogenetic analyses.     

Starch Consolidation Casting of Cordierite Precursor Mixtures—Rheological Behavior and Green Body Properties

The rheology of suspensions and mechanical properties of green bodies with cordierite composition (raw materials 37 wt% kaolin, 41 wt% talc, 22 wt% alumina, resulting in 46.6 wt% SiO₂, 38.1 wt% Al₂O₃, 13.6 wt% MgO) and two types of starch (corn or potato) are investigated. Rotational viscometry of suspensions with solids loading 50, 60, and 70 wt% without starch showed that all tend to be shear‐thinning with a small degree of thixotropy. Suspensions with a total solids loading of 60 wt% with 25 wt% replaced by starch exhibited higher viscosity and thixotropy, but the viscometric behavior is almost identical for the two starch types (apparent viscosities 130–50 mPa ·s). Oscillatory rheometry shows that for suspensions with potato starch the onset temperature for gelatinization is 61°C–63°C, that is, lower than for corn starch (72°C–73°C). Maximum storage moduli and phase shift values after gelatinization are similar for both systems. The mechanical properties of green disks, measured via diametral compression tests, reveal clear differences between materials prepared with corn and potato starch, with the latter showing higher elastic modulus, higher strength, and higher deformation at fracture, obviously because of incompletely gelatinized starch granules in the green bodies prepared with corn starch .     

Introducing a Semi-Coated Model to Investigate Antibacterial Effects of Biocompatible Polymers on Titanium Surfaces

Peri-implant infections from bacterial biofilms on artificial surfaces are a common threat to all medical implants. They are a handicap for the patient and can lead to implant failure or even life-threatening complications. New implant surfaces have to be developed to reduce biofilm formation and to improve the long-term prognosis of medical implants. The aim of this study was (1) to develop a new method to test the antibacterial efficacy of implant surfaces by direct surface contact and (2) to elucidate whether an innovative antimicrobial copolymer coating of 4-vinyl-N-hexylpyridinium bromide and dimethyl(2-methacryloyloxyethyl) phosphonate (VP:DMMEP 30:70) on titanium is able to reduce the attachment of bacteria prevalent in peri-implant infections. With a new in vitro model with semi-coated titanium discs, we were able to show a dramatic reduction in the adhesion of various pathogenic bacteria (Streptococcus sanguinis, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis), completely independently of effects caused by soluble materials. In contrast, soft tissue cells (human gingival or dermis fibroblasts) were less affected by the same coating, despite a moderate reduction in initial adhesion of gingival fibroblasts. These data confirm the hypothesis that VP:DMMEP 30:70 is a promising antibacterial copolymer that may be of use in several clinical applications.     

Degradation of Phenolic Compounds in Aqueous Sucrose Solutions by Ozonation

The sugar industry is concerned with color formation due to the oxidation of phenolic compounds in the presence of carbohydrates. In this study, we investigated the ozonation of a mixture of five phenolic compounds in water and aqueous sucrose solution: p-coumaric (p-COU), caffeic (CAF), syringic (SYR), and chlorogenic (CHO) acids, as well as the flavonoid quercetin (QUE). The experiments were carried out in a 3-L glass reactor with magnetic stirring and a diffuser plate at the bottom to feed the ozone-oxygen gas mixture. Initial solution concentrations of 5 mg L?1 of each acid, 15 mg L?1 of quercetin, and 40 g L?1 of sucrose were used. The degradation of phenolic compounds followed apparent first-order kinetics, with rate constants and percent removals decreasing in the presence of sucrose. In water, average consumed ozone dosages of 10.4 and 18.7 mg L?1 were necessary for 50% and 90% removals, respectively, for CHO, CAF, and p-COU; for QUE they were slightly higher (13.9 and 20.5 mg L?1, respectively). At a consumed dosage of 20.8 mg O3 L?1, more than 99% removal was obtained for CHO, CAF, and p-COU, while 96.2% was achieved for SYR. In contrast, QUE revealed to be more recalcitrant during ozonation in the absence of sucrose, with only 70% removal at the highest consumed O3 dosage. The consumed ozone dosages for 50% and 90% removals were higher for CHO, CAF, and p-COU in aqueous sucrose solution, which may impact ozone consumption during real sugarcane juice treatment. Sucrose and t-butanol were the main influential parameters that significantly affected the total amount of phenolic compounds degraded.     

Selective sorption of a Ge(IV) oxoanion by composite sorbent with hydrous oxide of cerium

In this work, composite sorbent based on hydrous cerium oxide was used for selective removal of a Ge (IV) oxoanion. Experiments were carried out by batch equilibrium tests and dynamic column sorption. The best sorption capacity of the Ge(IV) anion was reached at a pH of 9, where the sorption capacity was about 1.1 g/L. The negative effect of chlorides and sulfates was not observed in the concentration range of 100–1000 mg/L. The optimal flow rate for Ge(VI) sorption by sorbent CeO₂/XAD-7 was determined to be 6–12 BV/h. For regeneration, 10 BV of 1 mol/L HCl solution was used.     

Study of the Mass Transfer Behavior in a Centrifugal Contactor and Verification of the Solvent Extraction Model for the SX Process Program

The SX Process program has been developed for modelling of extraction processes in centrifugal contactors where the transfer kinetics is of big importance due to the short hold up time. Apparent distribution ratios are calculated using a stage efficiency which is flow-rate independent. In this work the dependency of the stage efficiency on parameters affecting the extraction transfer rate, such as metal loading, O/A ratio and acidity, has been investigated in single stage centrifugal contactor experiments for extraction of americium(III) into a 0.015 M CyMe₄-BTBP/0.25 M DMDOHEMA/octanol system. A model is proposed on how to calculate the stage efficiency and to accurately predict the apparent distribution ratios under the different conditions used.