Barrett’s Metaplasia Progression towards Esophageal Adenocarcinoma: An Attempt to Select a Panel of Molecular Sensors and to Reflect Clinical Alterations by Experimental Models

The molecular processes that predispose the development of Barrett’s esophagus (BE) towards esophageal adenocarcinoma (EAC) induced by gastrointestinal reflux disease (GERD) are still under investigation. In this study, based on a scientific literature screening and an analysis of clinical datasets, we selected a panel of 20 genes covering BE- and EAC-specific molecular markers (FZD5, IFNGR1, IL1A, IL1B, IL1R1, IL1RN, KRT4, KRT8, KRT15, KRT18, NFKBIL1, PTGS1, PTGS2, SOCS3, SOX4, SOX9, SOX15, TIMP1, TMEM2, TNFRSF10B). Furthermore, we aimed to reflect these alterations within an experimental and translational in vitro model of BE to EAC progression. We performed a comparison between expression profiles in GSE clinical databases with an in vitro model of GERD involving a BE cell line (BAR-T) and EAC cell lines (OE33 and OE19). Molecular responses of cells treated with acidified bile mixture (BM) at concentration of 100 and 250 μM for 30 min per day were evaluated. We also determined a basal mRNA expression within untreated, wild type cell lines on subsequent stages of BE and EAC development. We observed that an appropriately optimized in vitro model based on the combination of BAR-T, OE33 and OE19 cell lines reflects in 65% and more the clinical molecular alterations observed during BE and EAC development. We also confirmed previous observations that exposure to BM (GERD in vitro) activated carcinogenesis in non-dysplastic cells, inducing molecular alternations in the advanced stages of BE. We conclude that it is possible to induce, to a high extent, the molecular profile observed clinically within appropriately and carefully optimized experimental models, triggering EAC development. This experimental scheme and molecular marker panel might be implemented in further research, e.g., aiming to develop and evaluate novel compounds and prodrugs targeting GERD as well as BE and EAC prevention and treatment.     

The relationship between polypeptides and foaming during fermentation

The foam level during fermentations of hopped and unhopped wort and with fresh yeast and successive generations of yeast was examined. Simultaneously the total and hydrophobic polypeptide contents in worts, fermented worts and their concomitant foams were checked. It was shown that the total and hydrophobic polypeptide contents of the foam fraction and the foam level during fermentation were dependent on the generation number of yeast. The early generations of yeast (generations 1 and 2) promoted the formation of the largest amount of foam. It was also observed that a higher volume of foam occurred during fermentation of hopped wort in respect to unhopped one despite a higher concentration of polypeptides in unhopped wort. It could be a consequence of a higher foaming potential of polypeptides in hopped wort. The findings of the work may result in the limitation of foaming in the fermenters and consequently the increase of the brewery productivity without using additives and compromising the quality of the final product.     

Malted and unmalted oats in brewing

Using oats as a raw material in brewing has recently become the focus of increased interest. This is due to research findings that have shown that oats can be consumed safely by coeliac sufferers. It is also a response to consumer demand for products with novel sensory properties. In this study, beer was produced entirely from oat malt, from barley malt and from oat and barley malts mixed with various quantities of unmalted oats. Compared with barley wort, wort made from malted oats provided a lower extract content and had a higher protein content, but a lower free amino nitrogen content (FAN). The oat wort also showed increased viscosity and haze. The addition of unmalted oats during wort production produced significant changes in the physico‐chemical parameters of both oat and barley worts and beers. Unmalted oats caused an increase in wort viscosity and haze, and a reduction in total soluble nitrogen and FAN. Unmalted oats also contributed to lowering the concentration of higher alcohols and esters. Beer made from 100% oat and barley malts exhibited a similar alcohol content. The use of an oat adjunct in both cases resulted in a lower ethanol content. The introduction of enzyme preparations during the production of wort with oat adjunct had many benefits: increased extract content and FAN; a higher volume of wort; and a lower viscosity that led to faster wort filtration. This research suggests that the use of enzymes is necessary to make production using a high proportion of oats in the grist profitable. Copyright © 2014 The Institute of Brewing & Distilling     

Investigation of foam‐active polypeptides during beer fermentation

During ale fermentation there was an accumulation of total and hydrophobic polypeptides in the foam relative to the wort. Comparisons were made not only of the total and hydrophobic polypeptide contents but also of the molecular weights of these polypeptides present in wort, partially fermented wort and its concomitant foam. Wort, fermented wort and foam fractions had very similar polypeptide compositions with a major group having molecular weights of 40–43 kDa. Material of molecular weight in the range of 5–17 kDa and at 66 kDa was also detected. The polypeptides accumulated in foam displayed both hydrophobic and non‐hydrophobic character. The presence of yeast polypeptides in foam was confirmed. Comparison was also made between the fermentations of 10°Plato and 15°Plato wort. The results of the work may contribute to a better understanding of the mechanism of foam formation during beer fermentation, leading to reduced foaming and enabling an increase in the working capacities of fermenters. Copyright © 2004 Society of Chemical Industry     

Activated Carbon Modified with Different Chemical Agents as a Catalyst in the Dehydration and Dehydrogenation of Isopropanol

Activated carbons modified with different chemical agents such as HNO₃, H₂SO₄, peroxyacetic acid (PAA), air, NH₃ and Cl₂ have been tested as catalysts in decomposition (dehydration and dehydrogenation) of isopropanol. The majority of the samples obtained have been characterised by well-developed microporous surface with a small contribution of mesopores (8–18%). The influence of the surface area of the samples on their catalytic performance has been insignificant. The carbon oxidation with oxidants in the liquid or gas phase leads to an increased catalytic activity and the dominant process is dehydration of the alcohol studied. Carbon modification by contact with gas ammonia or chlorine results in a decrease in the catalytic activity and a significant increase in the contribution of dehydrogenation of isopropanol. It has been shown that such behaviour of the catalysts has been a consequence of changes in the acid-base character of the carbons induced by their modification.     

Isoprenoid Derivatives of Lysophosphatidylcholines Enhance Insulin and GLP-1 Secretion through Lipid-Binding GPCRs

Insulin plays a significant role in carbohydrate homeostasis as the blood glucose lowering hormone. Glucose-induced insulin secretion (GSIS) is augmented by glucagon-like peptide (GLP-1), a gastrointestinal peptide released in response to ingesting nutriments. The secretion of insulin and GLP-1 is mediated by the binding of nutrients to G protein-coupled receptors (GPCRs) expressed by pancreatic β-cells and enteroendocrine cells, respectively. Therefore, insulin secretagogues and incretin mimetics currently serve as antidiabetic treatments. This study demonstrates the potency of synthetic isoprenoid derivatives of lysophosphatidylcholines (LPCs) to stimulate GSIS and GLP-1 release. Murine insulinoma cell line (MIN6) and enteroendocrinal L cells (GLUTag) were incubated with LPCs bearing geranic acid (1-GA-LPC), citronellic acid (1-CA-LPC), 3,7-dimethyl-3-vinyloct-6-enoic acid (GERA-LPC), and (E)-3,7,11-trimethyl- 3-vinyldodeca-6,10-dienoic acid (1-FARA-LPC). Respective free terpene acids were also tested for comparison. Besides their insulin- and GLP-1-secreting capabilities, we also investigated the cytotoxicity of tested compounds, the ability to intracellular calcium ion mobilization, and targeted GPCRs involved in maintaining lipid and carbohydrate homeostasis. We observed the high cytotoxicity of 1-GERA-LPC and 1-FARA-LPC in contrast 1-CA-LPC and 1-GA-LPC. Moreover, 1-CA-LPC and 1-GA-LPC demonstrated the stimulatory effect on GSIS and 1-CA-LPC augmented GLP-1 secretion. Insulin and GLP-1 release appeared to be GPR40-, GPR55-, GPR119- and GPR120-dependent.     

Application of Conjugated Polymer–Platinum Group Metal Composites as Heterogeneous Catalysts

Composites of polyaniline (PANI) or polypyrrole (PPy) doped with chloride ions and Pt or Rh particles were prepared by chemical reduction of metal ions conducted in the presence of the polymers. Based on X-ray diffraction studies it was established that the composites contained metal nanoparticles (5–9 in size). However, according to SEM investigations metal particles were agglomerated (40 nm–1.1 μm in size). Redox activity of the composites in the catalytic isopropyl alcohol conversion was ca. ten times higher than the acid–base one. Pt dispersed in polymer matrices showed higher catalytic activity than Rh. PPy doped with chloride ions had a promoting effect on the activity of Pt catalysts.     

Numerical investigation of a high pressure hydrogen jet of 82 MPa with adaptive mesh refinement: Concentration and velocity distributions

To investigate the safety properties of high-pressure hydrogen discharge or leakage, an under-expanded hydrogen jet flow with a storage pressure of 82 MPa from a small jet orifice with a diameter of 0.2 mm is studied by three-dimensional (3D) numerical calculations. The full 3D compressible Navier-Stokes equations are utilized in a domain with a size of about 3 × 3 × 6 m which is discretized by employing an adaptive mesh refinement (AMR) technology to reduce the number of grid cells. By AMR, the local mesh resolutions can narrowly cover the Taylor microscale $l_T$ and direct numerical simulations (DNS) are performed. Both the instantaneous and mean hydrogen concentration distributions in the present jet are discussed. The instantaneous concentrations of hydrogen $C_{{\text{H}}_2}$ on the axis presents significant turbulent pulsating oscillations. The centerline value of the intensity of concentration fluctuation ${\stackrel{?}{\sigma }}_{{\text{H}}_2}$ asymptotically comes to 0.23, which is in a good agreement with the existing experimental results. It substantiates the conclusion that the asymptotic centerline value of ${\stackrel{?}{\sigma }}_{{\text{H}}_2}$ is independent of jet density ratio. The probability distributions function (PDF) of instantaneous axial $C_{{\text{H}}_2}$ agree approximately with the Gaussian distribution while skewing a little to the higher range. The time averaged hydrogen concentration ${\overline{C}}_{{\text{H}}_2}$ along the radial directions can also be described as a Gaussian distribution. The axial ${\overline{C}}_{{\text{H}}_2}$ of 82 MPa hydrogen jet tends to obey the distribution discipline approximated with ${\overline{C}}_{{\text{H}}_2}=4200/(z/\theta )$ where z is the axial distance from the nozzle and $\theta$ is the effective ejection diameter, which is consistent with the experimental results. In addition, the hydrogen tip penetration $Z_{\text{tip}}$ is found to be in a linear relationship with the square root of jet flow time $\sqrt{t}$. Meanwhile, the jet's velocity half-width $L_{\text{Vh}}$ approximately gains an linear relation with z which can be expressed as $L_{\text{Vh}}=0.09z$.     

New Glycosylated Dihydrochalcones Obtained by Biotransformation of 2′-Hydroxy-2-methylchalcone in Cultures of Entomopathogenic Filamentous Fungi

Flavonoids, including chalcones, are more stable and bioavailable in the form of glycosylated and methylated derivatives. The combined chemical and biotechnological methods can be applied to obtain such compounds. In the present study, 2′-hydroxy-2-methylchalcone was synthesized and biotransformed in the cultures of entomopathogenic filamentous fungi Beauveria bassiana KCH J1.5, Isaria fumosorosea KCH J2 and Isaria farinosa KCH J2.6, which have been known for their extensive enzymatic system and ability to perform glycosylation of flavonoids. As a result, five new glycosylated dihydrochalcones were obtained. Biotransformation of 2′-hydroxy-2-methylchalcone by B. bassiana KCH J1.5 resulted in four glycosylated dihydrochalcones: 2′-hydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside, 2′,3-dihydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside, 2′-hydroxy-2-hydroxymethyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside, and 2′,4-dihydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside. In the culture of I. fumosorosea KCH J2 only one product was formed—3-hydroxy-2-methyldihydrochalcone 2′-O-β-d-(4″-O-methyl)-glucopyranoside. Biotransformation performed by I. farinosa KCH J2.6 resulted in the formation of two products: 2′-hydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside and 2′,3-dihydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside. The structures of all obtained products were established based on the NMR spectroscopy. All products mentioned above may be used in further studies as potentially bioactive compounds with improved stability and bioavailability. These compounds can be considered as flavor enhancers and potential sweeteners.     

The switch mechanism of the cell death mode from apoptosis to necrosis in menadione-treated human osteosarcoma cell line 143B cells

Time-dependent changes in the cell death mode from apoptosis to necrosis were studied in cultured 143B cells treated with menadione, an anti-cancerous drug, excluding a possible involvement of "secondary necrosis." The population of apoptotic cells judged by FITC-Annexin V and propidium iodide (PI) double staining reached its maximum at 6 hours after 100 microM menadione treatment followed by an abrupt decrease thereafter, while that of necrotic cells continuously increased reaching 90% at 24 hours. Electron microscopically, cells attached to the culture dish at 6 hours after the treatment consisted of two different types of cells: cells with typical apoptotic features occupying the major population and those with condensed nuclei and swollen cytoplasm. Cells attached to the culture dish at 8 hours after the treatment consisted exclusively of those with condensed nuclei and swollen cytoplasm. Mitochondria in these cells showed various structural changes: those swollen to various degrees with deposition of flocculent densities, or those with highly condensed matrix. Distinct decreases both in intracellular levels of ATP and caspase-3-like activities and remarkable elevations of intracellular levels of superoxide, which were partly suppressed by NAD(P)H oxidase inhibitors, occurred at 6 hours after the treatment. These results may suggest that distinct increases of the intracellular level of superoxide derived from plasma membrane NAD(P)H oxidase besides that from mitochondria have triggered the transition of cell death mode from apoptosis to necrosis. Transition of highly condensed mitochondria to extremely swollen ones may reflect necrotic processes in menadione-treated cells. The present study strongly suggests that time-dependent study is essential using the electron microscopic technique to analyze detailed processes in the changes of the cell death mode.