Syndecan-1 Depletion Has a Differential Impact on Hyaluronic Acid Metabolism and Tumor Cell Behavior in Luminal and Triple-Negative Breast Cancer Cells

Glycosaminoglycans (GAGs) and proteoglycans (PGs) are major components of the glycocalyx. The secreted GAG and CD44 ligand hyaluronic acid (HA), and the cell surface PG syndecan-1 (Sdc-1) modulate the expression and activity of cytokines, chemokines, growth factors, and adhesion molecules, acting as critical regulators of tumor cell behavior. Here, we studied the effect of Sdc-1 siRNA depletion and HA treatment on hallmark processes of cancer in breast cancer cell lines of different levels of aggressiveness. We analyzed HA synthesis, and parameters relevant to tumor progression, including the stem cell phenotype, Wnt signaling constituents, cell cycle progression and apoptosis, and angiogenic markers in luminal MCF-7 and triple-negative MDA-MB-231 cells. Sdc-1 knockdown enhanced HAS-2 synthesis and HA binding in MCF-7, but not in MDA-MB-231 cells. Sdc-1-depleted MDA-MB-231 cells showed a reduced CD24-/CD44+ population. Furthermore, Sdc-1 depletion was associated with survival signals in both cell lines, affecting cell cycle progression and apoptosis evasion. These changes were linked to the altered expression of KLF4, MSI2, and miR-10b and differential changes in Erk, Akt, and PTEN signaling. We conclude that Sdc-1 knockdown differentially affects HA metabolism in luminal and triple-negative breast cancer model cell lines and impacts the stem phenotype, cell survival, and angiogenic factors.     

On-line laser-photometric monitoring of aerosol deposition in ventilated rabbit lungs

A photometric technique was developed for on-line measurement of aerosol deposition in isolated, ventilated, and perfused rabbit lungs. A jet nebulizer was used for aerosolization of saline (hygroscopic particles) and di(2-ethylhexyl) sebacate (nonhygroscopic particles). Aerosol concentration (laser photometer, constructed for measurements in rabbit lungs) and flow rate (commercial pneumotachograph) were continuously monitored at the inlet of the tracheal cannula. Computer-assisted data processing allowed the breath-by-breath calculation of inhaled and exhaled aerosol mass, thus providing the deposition fraction. With the use of hygroscopic particles, however, this approach was hampered by the humidity-induced particle growth in the airways, leading to an overestimation of the aerosol concentration in exhaled air. This effect was corrected by an algorithm using a "particle growth factor" derived breath by breath from the photometer signal. To test the reliability of this approach, saline particles carrying technetium-99m label were aerosolized into rabbit lungs with the use of various ventilator settings, and the aerosol deposition was assessed in parallel by photometry and by radioactivity detection over the lung and over a trap in the exhaled-air circuit. Superimposable curves of cumulative aerosol deposition, with changes in kinetics dependent on the ventilator mode, were obtained. For a given ventilator setting, absolute values of the deposition fraction were 0.32 +/- 0.04 (radiotracer quantification) and 0.36 +/- 0.04 (photometry; means +/- SD; n = 4). We conclude that the presented laser-photometric technique allows reliable on-line monitoring of the deposition of both nonhygroscopic and hygroscopic aerosol particles in ventilated lungs.     

The metabolism of fenazox in tomato plants

Results from investigations on the metabolism of 14C-Fenazox show that in 6-8 weeks old tomato-plants (sort "Harzfeuer") the agent undergoes a biotransformation. After chromatographic separation the structure of the biotransformation products was elucidated by comparison to authentic test substances, by derivatization as well as by means of physico-chemical methods. With that, the following metabolites were identified: non-transformed Fenazox, o-hydroxyazobenzene, o-hydroxyaniline, and p-hydroxyaniline. Probably, the agent is first transformed enzymatically or non-enzymatically into o-hydroxyazobenzene, then follows a reductive cleavage into o-hydroxyaniline and aniline. In its turn, the latter is transformed into o-hydroxy- and p-hydroxyaniline. Experiments in the field of transformation kinetics indicated that the concentration of the applied Fenazox rapidly decreases and that the first transformation product o-hydroxyazobenzene in the beginning increases and then again decreases in its concentration. With increasing application time it comes to an increase of the non-extractable residues.     

An Oral Mixed Fat Load Is Followed by a Modest Anti-inflammatory Adipocytokine Response in Overweight Patients with Metabolic Syndrome

We investigated the postprandial changes in plasma levels of adipocytokines in overweight patients with metabolic syndrome after an oral fat load. After an oral fat load and during a prolonged fast, blood was drawn at 0, 2, 3, 4 and 8 h for measurement of adiponectin, adipsin, cathepsin S, chemerin, hepatic growth factor, interferon‐γ‐inducible protein‐10, leptin, macrophage chemoattractant protein‐1, macrophage migration inhibitory factor, nerve growth factor, retinol binding protein‐4, resistin, serum amyloid A1, tissue inhibitor of metalloproteinase‐1 and thrombopoietin using a microbead‐based Luminex assay. Area under the curves (AUC) were calculated and compared. Plasma adiponectin levels were higher after an oral fat load compared to fasting at t = 2 h (950 ± 513 vs. ?1,881 ± 713 ng/ml) while the plasma levels for adipsin (?9 ± 5 vs. 16 ± 5 ng/ml), chemerin (?122 ± 35 vs. 13 ± 21 ng/ml), SAA‐1 (?391 ± 213 vs. 522 ± 173 ng/ml) and TPO (?335 ± 144 vs. 622 ± 216 ng/ml) were lower after an oral fat load compared to fasting. The baseline corrected AUC for IP‐10 was higher after fat load compared to fasting (median ?116 pg h/ml; IQR ?270 to 10 vs. ?21 pg h/ml; IQR ?136 to 418 (p = 0.047). In conclusion, in overweight male subjects with the metabolic syndrome, an oral fat load is accompanied with a modest anti‐inflammatory response of adipose tissue‐derived adipocytokines.     

Perceptions of the health and convenience characteristics of fresh and dried fruits

Dried fruits consumption has not received much research attention, although these foods are associated with potential health benefits. Therefore, the aim of this study was to compare fresh fruits and dried fruits on measures of health, convenience and consumer feelings.An on-line questionnaire that measured consumers' perceptions of the health benefits and convenience characteristics and their feelings about the consumption of both fresh fruits (apples and plums) and dried fruits (e.g., raisins, dates, and apricots) was completed by 693 respondents (Polish, French, and Dutch).Generally, fresh fruits were perceived to be healthier and less convenient than dried fruits. Consumers also reported more positive feelings about the consumption of fresh fruits compared to dried fruits. The more that the respondents were willing to sacrifice for their health, the more positively they rated the health aspects of both fresh and dried fruits, as well as several perceptions of convenience of both fresh and dried fruits. Health preoccupation was unrelated to most of the convenience perceptions and was associated with lower scores on several perceptions of healthiness. Recommendations for the consumer-oriented development of dried fruits are also provided.     

Gb3 Glycosphingolipids with Fluorescent Oligoene Fatty Acids: Synthesis and Phase Behavior in Model Membranes

Glycosphingolipids are involved in a number of physiological and pathophysiological processes, and they serve as receptors for a variety of bacterial toxins and viruses. To investigate their function in lipid membranes, fluorescently labeled glycosphingolipids are highly desirable. Herein, a synthetic route to access Gb₃ glycosphingolipids with fluorescently labeled fatty acids, consisting of pentaene and hexaene moieties either at the terminus or in the middle of the acyl chain, has been developed. The fluorescent properties of the Gb₃ derivatives were investigated in small unilamellar vesicles composed of a raft-like mixture. Phase-separated giant unilamellar vesicles (GUVs) allowed the quantification of the apparent partitioning coefficients of the Gb₃ compounds by means of confocal fluorescence laser scanning microscopy. The determined partition coefficients demonstrate that the Gb₃ derivatives are preferentially localized in the liquid-disordered (l_d) phase. To analyze whether the compounds behave like their physiological counterparts, Cy3-labeled (Cy: cyanine) Shiga toxin B subunits (STxB) were specifically bound to Gb₃-doped GUVs. However, the protein was favorably localized in the l_d phase, in contrast to results reported for STxB bound to naturally occurring Gb₃, which is discussed in terms of the packing density of the lipids in the liquid-ordered (l_o) phase.     

Efficient Engineering and Production Concepts for Products in Regulated Environments – Dream or Nightmare?

Manufacturing of chemical‐pharmaceutical products is moving increasingly fast on a global scale. Therefore, developing and starting up production facilities fast, with high quality, and at reasonable costs has become extremely challenging. Engineering concepts like modularization, standardization and simultaneous/parallel engineering are discussed as methods for speeding up process design and filing for regulatory approval. Transfer from batch to continuous operation mode of production is pointed out as the key‐issue in such strategies.