Analysis of microRNAs in Exosomes of Breast Cancer Patients in Search of Molecular Prognostic Factors in Brain Metastases

Brain metastases are the most severe tumorous spread during breast cancer disease. They are associated with a limited quality of life and a very poor overall survival. A subtype of extracellular vesicles, exosomes, are sequestered by all kinds of cells, including tumor cells, and play a role in cell-cell communication. Exosomes contain, among others, microRNAs (miRs). Exosomes can be taken up by other cells in the body, and their active molecules can affect the cellular process in target cells. Tumor-secreted exosomes can affect the integrity of the blood-brain barrier (BBB) and have an impact on brain metastases forming. Serum samples from healthy donors, breast cancer patients with primary tumors, or with brain, bone, or visceral metastases were used to isolate exosomes and exosomal miRs. Exosomes expressed exosomal markers CD63 and CD9, and their amount did not vary significantly between groups, as shown by Western blot and ELISA. The selected 48 miRs were detected using real-time PCR. Area under the receiver-operating characteristic curve (AUC) was used to evaluate the diagnostic accuracy. We identified two miRs with the potential to serve as prognostic markers for brain metastases. Hsa-miR-576-3p was significantly upregulated, and hsa-miR-130a-3p was significantly downregulated in exosomes from breast cancer patients with cerebral metastases with AUC: 0.705 and 0.699, respectively. Furthermore, correlation of miR levels with tumor markers revealed that hsa-miR-340-5p levels were significantly correlated with the percentage of Ki67-positive tumor cells, while hsa-miR-342-3p levels were inversely correlated with tumor staging. Analysis of the expression levels of miRs in serum exosomes from breast cancer patients has the potential to identify new, non-invasive, blood-borne prognostic molecular markers to predict the potential for brain metastasis in breast cancer. Additional functional analyzes and careful validation of the identified markers are required before their potential future diagnostic use.     

Molecular Cross-Talk between Gravity- and Light-Sensing Mechanisms in Euglena gracilis

Euglena gracilis is a photosynthetic flagellate. To acquire a suitable position in its surrounding aquatic environment, it exploits light and gravity primarily as environmental cues. Several physiological studies have indicated a fine-tuned relationship between gravity sensing (gravitaxis) and light sensing in E. gracilis. However, the underlying molecular mechanism is largely unknown. The photoreceptor photoactivated adenylyl cyclase (PAC) has been studied for over a decade. Nevertheless, no direct/indirect interaction partner (upstream/downstream) has been reported for PAC. It has been shown that a specific protein, kinase A (PKA), showed to be involved in phototaxis and gravitaxis. The current study reports the localization of the specific PKA and its relationship with PAC.     

Treatment of Critical Size Femoral Bone Defects with Biomimetic Hybrid Scaffolds of 3D Plotted Calcium Phosphate Cement and Mineralized Collagen Matrix

To treat critical-size bone defects, composite materials and tissue-engineered bone grafts play important roles in bone repair materials. The purpose of this study was to investigate the bone regenerative potential of hybrid scaffolds consisting of macroporous calcium phosphate cement (CPC) and microporous mineralized collagen matrix (MCM). Hybrid scaffolds were synthetized by 3D plotting CPC and then filling with MCM (MCM-CPC group) and implanted into a 5 mm critical size femoral defect in rats. Defects left empty (control group) as well as defects treated with scaffolds made of CPC only (CPC group) and MCM only (MCM group) served as controls. Eight weeks after surgery, micro-computed tomography scans and histological analysis were performed to analyze the newly formed bone, the degree of defect healing and the activity of osteoclasts. Mechanical stability was tested by 3-point-bending of the explanted femora. Compared with the other groups, more newly formed bone was found within MCM-CPC scaffolds. The new bone tissue had a clamp-like structure which was fully connected to the hybrid scaffolds and thereby enhanced the biomechanical strength. Together, the biomimetic hybrid MCM-CPC scaffolds enhanced bone defect healing by improved osseointegration and their differentiated degradation provides spatial effects in the process of critical-bone defect healing.     

From expert-driven to user-oriented communication of infection control guidelines

Currently, infection control guidelines in hospitals and other health care institutions are more expert-driven than user-oriented. In order to enhance the usability of the expert-driven guideline format, we developed a website for the communication of existing guidelines that better fit the practical information needs of health care workers (HCWs). We employed a user-centered design process that involved two studies.In the initial study, 28 HCWs were asked to solve tasks using existing, paper-based infection control guidelines. In order to detect their strategies and problems, the participants were asked to think aloud. Usability problems occurred due to poorly structured information, insufficient quality of information, and a mismatch between experts’ and HCWs’ vocabulary. To overcome these shortcomings, three design principles were applied for communicating infection control guidelines: better navigation (the guidelines should be searchable in several ways); multimodality (the guidelines should not be presented as text only), and action-orientation (the guidelines should be presented as HCWs’ behaviors). A website was developed to meet these principles.In the second study, the same 28 HCWs completed tasks identical to those of the first study while thinking aloud, but this time using the website. The percentage of correctly completed tasks increased and the mean time for task completion decreased significantly. Also, respondents were more satisfied with the website than the paper-based guidelines. The number of problems due to poor information quality and a mismatch in vocabulary declined, although the number of structural problems increased. This can probably be explained by the fact that the navigation structure was user-generated (using Card Sort), in contrast to a standardized answer format based on common usability principles.Overall, we found that involving HCWs in the development process is important to create a sense of ownership and to foster the implementation of the guidelines, which might eventually result in compliance and reduce health care-associated infections. This paper outlines concrete steps for how to involve HCWs in the design process.     

Tensile modulus modeling of carbon‐filled nylon 6,6 and polycarbonate‐based resins

Electrically and thermally conductive resins can be produced by adding conductive fillers to insulating polymers. Mechanical properties, such as tensile modulus, are also important. This research focused on performing compounding runs followed by injection molding and tensile testing of carbon‐filled nylon 6,6 and polycarbonate‐based resins. The three carbon fillers investigated included an electrically conductive carbon black, synthetic graphite particles, and a milled pitch–based carbon fiber. For each polymer, resins were produced and tested that contained varying amounts of these single‐carbon fillers. In addition, combinations of fillers were investigated by conducting a full 2³ factorial design and a complete replicate in each polymer. These tensile modulus experimental results were then compared to results predicted by several different models. For the composites containing only one filler type, the Nielsen model with the modified Ψ term provided the best prediction of the actual experimental values. For the composites containing more than one filler type, a new parameter, which includes the vibrated bulk density (VBD) of the fillers, was incorporated into the Nielsen model with the modified Ψ term. This model with the new VBD parameter provided the best estimate of experimental tensile modulus for composites containing multiple‐filler types. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 1716–1728, 2003     

Customizing the comonomer incorporation distribution in Ziegler–Natta-based LLDPE: Harnessing the influences of the titanation temperature during catalyst synthesis and of polymerization process parameters

The microstructure of linear low-density polyethylene (LLDPE) is strongly influenced by short-chain branches (SCBs) incorporated into the polymer backbone. Varying the number, distribution, and length of SCBs allows the properties of the resulting polymer to be tailored to meet specific requirements. Using Ziegler–Natta (ZN) catalysts for synthesis has disadvantages in terms of the comonomer incorporation distribution (CID) compared to, for instance, metallocene and post–metallocene catalysts. Nevertheless, ZN catalysts continue to be widely used, as many of the new generations of catalysts are more difficult to handle and cannot match the cheap cost of ZN catalysts. To improve this aspect of ZN catalysts, we investigated the influence of catalyst titanation temperature and polymerization process parameters on the CID. Our results show that it is possible to manipulate the process parameters of the present ZN catalyst system to yield a desired comonomer amount and CID in the polymer. Varying the titanation temperature clearly influenced the titanium content of the catalyst. Molecular-weight distribution analysis and deconvolution results indicate that changes in the amounts of comonomer incorporated and in the CID are directly related to the catalyst's active site that produces the lowest-molecular-weight fraction.     

Novel Type II Fatty Acid Biosynthesis (FAS II) Inhibitors as Multistage Antimalarial Agents

Malaria is a potentially fatal disease caused by Plasmodium parasites and poses a major medical risk in large parts of the world. The development of new, affordable antimalarial drugs is of vital importance as there are increasing reports of resistance to the currently available therapeutics. In addition, most of the current drugs used for chemoprophylaxis merely act on parasites already replicating in the blood. At this point, a patient might already be suffering from the symptoms associated with the disease and could additionally be infectious to an Anopheles mosquito. These insects act as a vector, subsequently spreading the disease to other humans. In order to cure not only malaria but prevent transmission as well, a drug must target both the blood‐ and pre‐erythrocytic liver stages of the parasite. P. falciparum (Pf) enoyl acyl carrier protein (ACP) reductase (ENR) is a key enzyme of plasmodial type II fatty acid biosynthesis (FAS II). It has been shown to be essential for liver‐stage development of Plasmodium berghei and is therefore qualified as a target for true causal chemoprophylaxis. Using virtual screening based on two crystal structures of PfENR, we identified a structurally novel class of FAS inhibitors. Subsequent chemical optimization yielded two compounds that are effective against multiple stages of the malaria parasite. These two most promising derivatives were found to inhibit blood‐stage parasite growth with IC₅₀ values of 1.7 and 3.0 μM and lead to a more prominent developmental attenuation of liver‐stage parasites than the gold‐standard drug, primaquine.     

A Nondestructive Methodology for the Study of Colored Enamels: Insights into Manufacturing and Weathering Processes

We studied ancient enamels on gilded copper from a collection of archeological horse harness pendants of the Museo Instituto Valencia de Don Juan (Madrid, Spain) to test the benefits of a new, nondestructive analytical methodology based on chemometric analysis (i.e., Principal Component Analysis, PCA) on micro‐ATR‐FTIR spectral data and chemical quantification using SEM‐EDS. The novelty of this approach was threefold: (i) PCA allowed the discrimination of the different harness pendants of known origin and attributed to the 14th and 15th centuries according to the chemical complex composition, nanostructure, glass weathering, and/or coloring mechanisms of each colored enamel, separately (i.e., red, purple, blue, and white), (ii) it is a cheap, easily available and nondestructive methodology that enables us to (iii) draw archeological conclusions about the quality of the manufacturing process, reassess the chronology of these objects and attempt to attribute them to different workshops according to the different traditional recipes identified. In particular, the enamels were made of alkali and/or alkaline earth lead‐glass with a wide range of chemical compounds in the form of pigments or opacifiers. Two types of coloring mechanisms were identified, colloidal particles such as copper‐ruby for red enamels, and ionic mechanisms such as Fe(II) and Co(II) to achieve a blue pigments; Mn(III) in the purple pigment; and two kind of white enamels were identified, i.e., tin oxide as an opacifier and uranium oxide. In addition, we established the reason for the poor state of conservation of some of the enamels by means of the identification of depolymerization and ion exchanges, well‐known harmful effects of glass weathering, and finally a chronology was assigned for some of these pieces according to the enamel composition.     

Influence of precursors on the formation of 3‐MCPD and glycidyl esters in a model oil under simulated deodorization conditions

To find new ways for reducing the potential of palm oil to form 3‐monochloropropane‐1,2‐diol (3‐MCPD) and glycidyl esters during refining it is helpful to know more about the influence of different precursors like diacylglycerols (DAGs) and monoacylglycerols (MAGs), lecithin, and chlorine containing compounds. After adding increasing amounts of the different precursors to a model oil obtained by removal of polar compounds from crude palm oil and heating the mixture under standardized conditions to 240°C for 2 h the contents of 3‐MCPD and glycidyl esters were analyzed according to the standard procedure of DGF C‐VI 18 (10). DAGs and MAGs were found to increase the potential of palm oil to form 3‐MCPD and glycidyl esters, but refined lecithin showed no influence. Sodium chloride as well as tetra‐n‐butylammoniumchloride (TBAC) led to higher contents of the esters. Whereas the addition of TBAC raised the amount of glycidyl esters as well as 3‐MCPD esters, sodium chloride largely raised the amount of 3‐MCPD esters. An addition of 5 mmol of sodium carbonate/kg model oil spiked with sodium chloride reduced the amount of glycidyl esters almost completely; the 3‐MCPD esters were reduced by 50%. About 1 mmol sodium hydrogen carbonate/kg oil reduced both 3‐MCPD and glycidyl esters almost completely. Practical applications : For the mitigation of the formation of 3‐MCPD esters and related compounds in refined edible oils, it is helpful to know more about the effect of different possible precursors. Using a broader data basis, it is possible to adopt the oil processing but especially the choice of the raw material to the demands of the market for lower contents of the esters in the refined products.