Spatially selective deposition of electrically charged microparticles onto integrated circuits that generate electrical fields in programmable patterns using electrodes on their surface was ...previously limited to a pixel pitch of 100
μm. Now, we demonstrate spatially selective deposition onto pixels of 45
μm pitch in experiments on a test chip allowing arbitrary patterns, but being of limited size and of fixed characteristics, complemented by COMSOL simulations. Experiments on a prototype high voltage CMOS chip demonstrate the feasibility of miniaturisation in the first place, imply simulations of interest that cannot be tested experimentally and, conversely, complement the simplified simulation models by reality checks. Using COMSOL for the optimisation of the setup parameters, particles of decreasing average diameter in a number of aerosol and electrical field geometries are simulated with particular attention to minimising contamination (deposition of particles on undesirable locations). Combining these results, the average particle diameter is decreased from 10
μm to less than 3
μm and the deposition voltage is reduced from 100
V to 30
V, when using pixels with a pitch of 45
μm. Optimising these parameters allows for more than quadrupling the spot density compared to the previous chip, on which combinatorial particle deposition with minimal contamination is achieved. Peptide arrays, having been previously shown to be a major application for this method, benefit in particular, as the increase in density from 10,000
pixels/cm
2 to approximately 50,000
pixels/cm
2 promises a significant decrease in cost-per-peptide and amount of test specimens required.
The behavior of charged bio polymer micro particles when deposited onto a CMOS chip can be analytically modeled in form of the incompressible Navier-Stokes equation and the electrostatic Poisson ...equation, as we describe in this article. Based on these models, numerical simulations of depositions can be implemented in COMSOL that lead to improvements in the experimental setup, optimizing the size and charge distribution of the micro particles. Adapting the experiments according to the simulation results, we will show the powerful gain in deposition precision, which is essential for a contamination-free deposition and hence high quality combinatorial deposition.
While insulin replacement therapy restores the health and prevents the onset of diabetic complications (DC) for many decades, some T1D patients have elevated hemoglobin A1c values suggesting poor ...glycemic control, a risk factor of DC. We surveyed the stool microbiome and urinary proteome of a cohort of 220 adolescents and children, half of which had lived with T1D for an average of 7 years and half of which were healthy siblings. Phylogenetic analysis of the 16S rRNA gene did not reveal significant differences in gut microbial alpha-diversity comparing the two cohorts. The urinary proteome of T1D patients revealed increased abundances of several lysosomal proteins that correlated with elevated HbA1c values.
protein network analysis linked such proteins to extracellular matrix components and the glycoprotein LRG1. LRG1 is a prominent inflammation and neovascularization biomarker. We hypothesize that these changes implicate aberrant glycation of macromolecules that alter lysosomal function and metabolism in renal tubular epithelial cells, cells that line part of the upper urinary tract.
We built high voltage complementary metal oxide semiconductor (CMOS) chips that generate electrical fields on their surface, such that electrically charged microparticles (diameter 10–20
μm on ...average) can be addressed on distinct pixel electrodes according to arbitrary field patterns. Each pixel contains a memory cell in canonical low-voltage CMOS-technology controlling a high voltage (30–100
V) potential area on the top metal layer. Particle transfer with minimal contaminations in less than 10
s for a complete chip was observed for pixels of 100
μm
×
100
μm down to 65
μm
×
65
μm. This allows a new way to create surface modifications on top of CMOS chips without need for additional masks or stamps. Using suitable particles, a chemically modified chip surface, and compatible chemistry, this method can be utilized for self-aligned high-density biopolymer arrays, e.g., peptide arrays. Transfer of microparticles loaded with amino acids for combinatorial peptide synthesis is demonstrated. Successful synthesis of different peptides (octamers) was proven by immunostaining. Based on results obtained by a chip containing pixel areas of different characteristics, a chip for biological applications with 16,384
pixels (10,000
pixel/cm
2) was built. Good homogeneity of peptide synthesis over the chip area was verified by immunostaining.
Individuals with type 1 diabetes (T1D) often have higher than normal blood glucose levels, causing advanced glycation end product formation and inflammation and increasing the risk of vascular ...complications years or decades later. To examine the urinary proteome in juveniles with T1D for signatures indicative of inflammatory consequences of hyperglycemia, we profiled the proteome of 40 T1D patients with an average of 6.3 years after disease onset and normal or elevated HbA1C levels, in comparison with a cohort of 41 healthy siblings. Using shotgun proteomics, 1036 proteins were identified, on average, per experiment, and 50 proteins showed significant abundance differences using a Wilcoxon signed-rank test (FDR q-value ≤ 0.05). Thirteen lysosomal proteins were increased in abundance in the T1D versus control cohort. Fifteen proteins with functional roles in vascular permeability and adhesion were quantitatively changed, including CD166 antigen and angiotensin-converting enzyme 2. α-N-Acetyl-galactosaminidase and α-fucosidase 2, two differentially abundant lysosomal enzymes, were detected in western blots with often elevated quantities in the T1D versus control cohort. Increased release of proteins derived from lysosomes and vascular epithelium into urine may result from hyperglycemia-associated inflammation in the kidney vasculature.
Context: As part of the third Gaia Data Release, we present the contributions of the non-stellar and classification modules from the eighth coordination unit (CU8) of the Data Processing and Analysis ...Consortium, which is responsible for the determination of source astrophysical parameters using Gaia data. This is the third in a series of three papers describing the work done within CU8 for this release.
Aims: For each of the five relevant modules from CU8, we summarise their objectives, the methods they employ, their performance, and the results they produce for Gaia DR3. We further advise how to use these data products and highlight some limitations.
Methods: The Discrete Source Classifier (DSC) module provides classification probabilities associated with five types of sources: quasars, galaxies, stars, white dwarfs, and physical binary stars. A subset of these sources are processed by the Outlier Analysis (OA) module, which performs an unsupervised clustering analysis, and then associates labels with the clusters to complement the DSC classification. The Quasi Stellar Object Classifier (QSOC) and the Unresolved Galaxy Classifier (UGC) determine the redshifts of the sources classified as quasar and galaxy by the DSC module. Finally, the Total Galactic Extinction (TGE) module uses the extinctions of individual stars determined by another CU8 module to determine the asymptotic extinction along all lines of sight for Galactic latitudes |b| > 5 degrees.
Results: Gaia DR3 includes 1591 million sources with DSC classifications; 56 million sources to which the OA clustering is applied; 1.4 million sources with redshift estimates from UGC; 6.4 million sources with QSOC redshift; and 3.1 million level 9 HEALPixes of size 0 :013 deg(2) where the extinction is evaluated by TGE.
Conclusions: Validation shows that results are in good agreement with values from external catalogues; for example 90% of the QSOC redshifts have absolute error lower than 0:1 for sources with empty warning flags, while UGC redshifts have a mean error of 0:008 +/- 0:037 if evaluated on a clean set of spectra. An internal validation of the OA results further shows that 30 million sources are located in high confidence regions of the clustering map.
Chronic liver disease due to alcohol-use disorder contributes markedly to the global burden of disease and mortality
. Alcoholic hepatitis is a severe and life-threatening form of alcohol-associated ...liver disease. The gut microbiota promotes ethanol-induced liver disease in mice
, but little is known about the microbial factors that are responsible for this process. Here we identify cytolysin-a two-subunit exotoxin that is secreted by Enterococcus faecalis
-as a cause of hepatocyte death and liver injury. Compared with non-alcoholic individuals or patients with alcohol-use disorder, patients with alcoholic hepatitis have increased faecal numbers of E. faecalis. The presence of cytolysin-positive (cytolytic) E. faecalis correlated with the severity of liver disease and with mortality in patients with alcoholic hepatitis. Using humanized mice that were colonized with bacteria from the faeces of patients with alcoholic hepatitis, we investigated the therapeutic effects of bacteriophages that target cytolytic E. faecalis. We found that these bacteriophages decrease cytolysin in the liver and abolish ethanol-induced liver disease in humanized mice. Our findings link cytolytic E. faecalis with more severe clinical outcomes and increased mortality in patients with alcoholic hepatitis. We show that bacteriophages can specifically target cytolytic E. faecalis, which provides a method for precisely editing the intestinal microbiota. A clinical trial with a larger cohort is required to validate the relevance of our findings in humans, and to test whether this therapeutic approach is effective for patients with alcoholic hepatitis.
Aerococcus urinae (Au) and Globicatella sanguinis (Gs) are gram-positive bacteria belonging to the family Aerococcaceae and colonize the human immunocompromised and catheterized urinary tract. We ...identified both pathogens in polymicrobial urethral catheter biofilms (CBs) with a combination of 16S rDNA sequencing, proteomic analyses, and microbial cultures. Longitudinal sampling of biofilms from serially replaced catheters revealed that each species persisted in the urinary tract of a patient in cohabitation with 1 or more gram-negative uropathogens. The Gs and Au proteomes revealed active glycolytic, heterolactic fermentation, and peptide catabolic energy metabolism pathways in an anaerobic milieu. A few phosphotransferase system (PTS)–based sugar uptake and oligopeptide ABC transport systems were highly expressed, indicating adaptations to the supply of nutrients in urine and from exfoliating squamous epithelial and urothelial cells. Differences in the Au vs Gs metabolisms pertained to citrate lyase and utilization and storage of glycogen (evident only in Gs proteomes) and to the enzyme Xfp that degrades d-xylulose-5′-phosphate and the biosynthetic pathways for 2 protein cofactors, pyridoxal 6′-phosphate and 4′-phosphopantothenate (expressed only in Au proteomes). A predicted ZnuA-like transition metal ion uptake system was identified for Gs while Au expressed 2 LPXTG-anchored surface proteins, one of which had a predicted pilin D adhesion motif. While these proteins may contribute to fitness and virulence in the human host, it cannot be ruled out that Au and Gs fill a niche in polymicrobial biofilms without being the direct cause of injury in urothelial tissues.
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