Fabrication of a biocompatible flexible electroosmosis micropump

In in-vivo microsystems, one of the components is a biocompatible micropump in order to produce the necessary force to deliver the fluid from the inlet to the outlet. In this contribution, a flexible micropump is fabricated which is aimed to be suitable in drug delivery applications. It provides high degree of biocompatibility, since the only employed materials are implantation grade polydimethylsiloxane elastomer and gold for the electrical interconnects. The working principle of the micropump is based on transverse DC electroosmosis which is a new variant of conventionally applied high voltage DC electroosmosis. This new technique is based on topography irregularities introduced in the channel resulting in a non-uniform charge distribution. The advantage is to drive the micropump using a relatively low DC voltage of 10 V while getting an effective flow speed of 60 μm/s. In order to characterize the flow speed, dyed 3 μm beads are dispersed in the working fluid and their speed is measured by the line scanning technique using a confocal microscope. It is also observed that the flow has a helical profile which is an attractive feature for an efficient micro-mixer in active microfluidics and μ-TAS applications.     

Notch sensitivity of thermoset and thermoplastic laminates loaded in tension

Inplane tensile fracture of unnotched and notched thermoset graphite-epoxy and thermoplastic graphite-PEEK composite laminates is examined. Both fibre-dominated quasi-isotropic and matrix dominated ±45 angle-ply layups were investigated.Classical lamination theory predictions of elastic and strength properties of unnotched specimens are compared with experiments. Several notched geometries, i.e. centre-notched, double-edge notched and open-hole specimens subjected to tensile loading to fracture were examined. The notched strength of the quasi-isotropic laminates was analysed by a damage zone model, where damage around the notch is represented by an equivalent crack with cohesive force acting between the crack surfaces.Good agreement between experimental and calculated strength was observed for the graphite-epoxy laminates which failed in a collinear manner. For the graphite-PEEK laminates discrepancies between predicted and experimental strength are related to observed deviations from collinear crack growth. The angle-ply graphite-PEEK laminates showed larger notch sensitivity than the corresponding graphite-epoxy, probably due to less degree of stress relieving damage formation around the notch.     

The effect of hydrothermic treatment on the physicochemical properties of rye grain

Summary The gelatinized, autoclaved and sodiumhydroxide-treated starch preparations were used and their interaction with soluble pentosans and gliadin was analysed. The molecular sieving technique and optical methods were used to detect the complexes formed. The modified starches formed complexes with both soluble pentosans and gliadin in acidic and neutral media (non-ionic molecular forces being involved). The autoclaved starch showed the highest reactivity. The amylose and amylopectin played different roles in the interactions and the aggregates of amylose with gliadin and amylopectin with soluble pentosans were the prevailing forms present in the complexes. Small quantities of other products were also identified.

---

Einfluß der hydrothermischen Behandlung auf die physiko-chemischen Eigenschaften des RoggenkornsII. Wechselwirkung der Eiweiß-und Kohlenhydratkomplexe (Modelluntersuchung)

Zusammenfassung Die Wechselwirkungen verkleisterter, autoklavierter und mit Natronlauge behandelter Stärke mit löslichen Pentosanen und Gliadin werden mit Hilfe der Gelpermeation und mit optischen Methoden untersucht. Die modifizierten Stärken bildeten Komplexe mit den Pentosanen und mit Gliadin im sauren und neutralen Medium, autoklavierte Stärke zeigte die hächste Reaktion. Die unterschiedliche Rolle von Amylose und Amylopection zeigte vorherrschend Amylose-Gliadin und Amylopectin-PentosanKomplexe, während andere Komplexe in kleineren Mengen auftraten.

     

The Effect of Hypoxia on the Expression of CXC Chemokines and CXC Chemokine Receptors—A Review of Literature

Hypoxia is an integral component of the tumor microenvironment. Either as chronic or cycling hypoxia, it exerts a similar effect on cancer processes by activating hypoxia-inducible factor-1 (HIF-1) and nuclear factor (NF-κB), with cycling hypoxia showing a stronger proinflammatory influence. One of the systems affected by hypoxia is the CXC chemokine system. This paper reviews all available information on hypoxia-induced changes in the expression of all CXC chemokines (CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8 (IL-8), CXCL9, CXCL10, CXCL11, CXCL12 (SDF-1), CXCL13, CXCL14, CXCL15, CXCL16, CXCL17) as well as CXC chemokine receptors—CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CXCR7 and CXCR8. First, we present basic information on the effect of these chemoattractant cytokines on cancer processes. We then discuss the effect of hypoxia-induced changes on CXC chemokine expression on the angiogenesis, lymphangiogenesis and recruitment of various cells to the tumor niche, including myeloid-derived suppressor cells (MDSCs), tumor-associated macrophages (TAMs), tumor-associated neutrophils (TANs), regulatory T cells (T_regs) and tumor-infiltrating lymphocytes (TILs). Finally, the review summarizes data on the use of drugs targeting the CXC chemokine system in cancer therapies.     

Sensitivity to Cisplatin in Head and Neck Cancer Cells Is Significantly Affected by Patient-Derived Cancer-Associated Fibroblasts

Cancer-associated fibroblasts (CAFs) are one of the most abundant and critical components of the tumor stroma. CAFs can impact many important steps of cancerogenesis and may also influence treatment resistance. Some of these effects need the direct contact of CAFs and cancer cells, while some involve paracrine signals. In this study, we investigated the ability of head and neck squamous cell carcinomas (HNSCC) patient-derived CAFs to promote or inhibit the colony-forming ability of HNSCC cells. The effect of cisplatin on this promoting or inhibiting influence was also studied. The subsequent analysis focused on changes in the expression of genes associated with cancer progression. We found that cisplatin response in model HNSCC cancer cells was modified by coculture with CAFs, was CAF-specific, and different patient-derived CAFs had a different “sensitizing ratio”. Increased expression of VEGFA, PGE2S, COX2, EGFR, and NANOG in cancer cells was characteristic for the increase of resistance. On the other hand, CCL2 expression was associated with sensitizing effect. Significantly higher amounts of cisplatin were found in CAFs derived from patients who subsequently experienced a recurrence. In conclusion, our results showed that CAFs could promote and/or inhibit colony-forming capability and cisplatin resistance in HNSCC cells via paracrine effects and subsequent changes in gene expression of cancer-associated genes in cancer cells.     

Energy Metabolites as Biomarkers in Ischemic and Dilated Cardiomyopathy

With more than 25 million people affected, heart failure (HF) is a global threat. As energy production pathways are known to play a pivotal role in HF, we sought here to identify key metabolic changes in ischemic- and non-ischemic HF by using a multi-OMICS approach. Serum metabolites and mRNAseq and epigenetic DNA methylation profiles were analyzed from blood and left ventricular heart biopsy specimens of the same individuals. In total we collected serum from n = 82 patients with Dilated Cardiomyopathy (DCM) and n = 51 controls in the screening stage. We identified several metabolites involved in glycolysis and citric acid cycle to be elevated up to 5.7-fold in DCM (p = 1.7 × 10⁻⁶). Interestingly, cardiac mRNA and epigenetic changes of genes encoding rate-limiting enzymes of these pathways could also be found and validated in our second stage of metabolite assessment in n = 52 DCM, n = 39 ischemic HF and n = 57 controls. In conclusion, we identified a new set of metabolomic biomarkers for HF. We were able to identify underlying biological cascades that potentially represent suitable intervention targets.     

GCase and LIMP2 Abnormalities in the Liver of Niemann Pick Type C Mice

The lysosomal storage disease Niemann–Pick type C (NPC) is caused by impaired cholesterol efflux from lysosomes, which is accompanied by secondary lysosomal accumulation of sphingomyelin and glucosylceramide (GlcCer). Similar to Gaucher disease (GD), patients deficient in glucocerebrosidase (GCase) degrading GlcCer, NPC patients show an elevated glucosylsphingosine and glucosylated cholesterol. In livers of mice lacking the lysosomal cholesterol efflux transporter NPC1, we investigated the expression of established biomarkers of lipid-laden macrophages of GD patients, their GCase status, and content on the cytosol facing glucosylceramidase GBA2 and lysosomal integral membrane protein type B (LIMP2), a transporter of newly formed GCase to lysosomes. Livers of 80-week-old Npc1^−/− mice showed a partially reduced GCase protein and enzymatic activity. In contrast, GBA2 levels tended to be reciprocally increased with the GCase deficiency. In Npc1^−/− liver, increased expression of lysosomal enzymes (cathepsin D, acid ceramidase) was observed as well as increased markers of lipid-stressed macrophages (GPNMB and galectin-3). Immunohistochemistry showed that the latter markers are expressed by lipid laden Kupffer cells. Earlier reported increase of LIMP2 in Npc1^−/− liver was confirmed. Unexpectedly, immunohistochemistry showed that LIMP2 is particularly overexpressed in the hepatocytes of the Npc1^−/− liver. LIMP2 in these hepatocytes seems not to only localize to (endo)lysosomes. The recent recognition that LIMP2 harbors a cholesterol channel prompts the speculation that LIMP2 in Npc1^−/− hepatocytes might mediate export of cholesterol into the bile and thus protects the hepatocytes.     

Refining Genotypes and Phenotypes in KCNA2-Related Neurological Disorders

Pathogenic variants in KCNA2, encoding for the voltage-gated potassium channel K_v1.2, have been identified as the cause for an evolving spectrum of neurological disorders. Affected individuals show early-onset developmental and epileptic encephalopathy, intellectual disability, and movement disorders resulting from cerebellar dysfunction. In addition, individuals with a milder course of epilepsy, complicated hereditary spastic paraplegia, and episodic ataxia have been reported. By analyzing phenotypic, functional, and genetic data from published reports and novel cases, we refine and further delineate phenotypic as well as functional subgroups of KCNA2-associated disorders. Carriers of variants, leading to complex and mixed channel dysfunction that are associated with a gain- and loss-of-potassium conductance, more often show early developmental abnormalities and an earlier onset of epilepsy compared to individuals with variants resulting in loss- or gain-of-function. We describe seven additional individuals harboring three known and the novel KCNA2 variants p.(Pro407Ala) and p.(Tyr417Cys). The location of variants reported here highlights the importance of the proline(405)–valine(406)–proline(407) (PVP) motif in transmembrane domain S6 as a mutational hotspot. A novel case of self-limited infantile seizures suggests a continuous clinical spectrum of KCNA2-related disorders. Our study provides further insights into the clinical spectrum, genotype–phenotype correlation, variability, and predicted functional impact of KCNA2 variants.     

X-ray Fluorescence Uptake Measurement of Functionalized Gold Nanoparticles in Tumor Cell Microsamples

Quantitative cellular in vitro nanoparticle uptake measurements are possible with a large number of different techniques, however, all have their respective restrictions. Here, we demonstrate the application of synchrotron-based X-ray fluorescence imaging (XFI) on prostate tumor cells, which have internalized differently functionalized gold nanoparticles. Total nanoparticle uptake on the order of a few hundred picograms could be conveniently observed with microsamples consisting of only a few hundreds of cells. A comparison with mass spectroscopy quantification is provided, experimental results are both supported and sensitivity limits of this XFI approach extrapolated by Monte-Carlo simulations, yielding a minimum detectable nanoparticle mass of just 5 pg. This study demonstrates the high sensitivity level of XFI, allowing non-destructive uptake measurements with very small microsamples within just seconds of irradiation time.