Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR

NMR flow devices provide longitudinal real-time quantitative metabolome characterisation of living cells. However, discrimination of intra- and extracellular contributions to the spectra represents a major challenge in metabolomic NMR studies. The present NMR study demonstrates the possibility to quantitatively measure both metabolic intracellular fingerprints and extracellular footprints on human control fibroblasts by using a commercially available flow tube system with a standard 5 mm NMR probe. We performed a comprehensive 3D cell culture system characterisation. Diffusion NMR was employed for intra- and extracellular metabolites separation. In addition, complementary extracellular footprints were determined. The implemented perfused NMR bioreactor system allowed the determination of 35 metabolites and intra- and extracellular separation of 19 metabolites based on diffusion rate differences. We show the reliability and sensitivity of NMR diffusion measurements to detect metabolite concentration changes in both intra- and extracellular compartments during perfusion with different selective culture media, and upon complex I inhibition with rotenone. We also demonstrate the sensitivity of extracellular footprints to determine metabolic variations at different flow rates. The current method is of potential use for the metabolomic characterisation of defect fibroblasts and for improving physiological comprehension.     

Electrodeposition of copper into high aspect ratio PCB micro-via using megasonic agitation

This paper presents the use of micro-particle imaging velocimetry (micro-PIV) to analyse fluid flow and hence ion replenishment in PCB micro-via during the electroplating process. The cross section of a PCB via is fabricated in PMMA to allow optical access to the sample. Fluid flow within two 1:1 aspect ratio blind micro-vias, one with straight side walls and the other with tapered side walls were compared. Flow is also analysed in a 1:1 aspect ratio through via. Flow rates measured using micro-PIV are used to validate simulated flow models. The results show that there are increased flow rates within the blind via with tapered side walls. This goes some way to explaining the improved electroplating results obtained in industry when tapered vias are used. Initial experimental results using megasonic streaming to remove bubbles from blind micro-via and promote ion transportation within high aspect ratio PCB micro-via to enhance electrodeposition are also reported.     

DNA Break Mapping Reveals Topoisomerase II Activity Genome-Wide

Genomic DNA is under constant assault by endogenous and exogenous DNA damaging agents. DNA breakage can represent a major threat to genome integrity but can also be necessary for genome function. Here we present approaches to map DNA double-strand breaks (DSBs) and single-strand breaks (SSBs) at the genome-wide scale by two methods called DSB- and SSB-Seq, respectively. We tested these methods in human colon cancer cells and validated the results using the Topoisomerase II (Top2)-poisoning agent etoposide (ETO). Our results show that the combination of ETO treatment with break-mapping techniques is a powerful method to elaborate the pattern of Top2 enzymatic activity across the genome.     

Analysis of electromigration phenomenon in thick-film and LTCC structures at elevated temperature

Studies on electromigration phenomenon in thick-film structures on alumina and LTCC substrates are presented in this paper. The effects of storage of Au and Ag electrode patterns in temperature range up to 300 °C under voltage bias were examined. The leakage characteristics of electrodes with 100 μm spacing at 50 V dc bias as a function of time and temperature are presented and analyzed. Scanning electron microscope (SEM) equipped with the energy-dispersive X-ray spectroscopy (EDX) detector was applied for determination of metal ions transport. Test structures with Au-based conductive material are much more resistant to electromigration than Ag-based layers.     

Microbial growth,communities and sensory characteristics of vacuum and modified atmosphere packaged lamb shoulders

Packaging fresh lamb in a vacuum (VAC) versus a 100% CO₂ modified atmosphere (MAP) may influence product shelf-life and the bacterial communities. While VAC is a common packing method and 100% CO₂ MAP is used in some countries, there is little information about how these different techniques affect the growth of spoilage bacteria and sensory attributes of lamb. The aim of this study was to assess changes in microbiological and organoleptic properties, and determine differences in microbial communities by terminal restriction fragment length polymorphism (TRFLP) and 454 pyrosequencing, in bone-in (BI) and bone-out (BO) MAP- and VAC-packed lamb shoulders stored at −0.3 °C over 12 wk. VAC and MAP lamb shoulders were acceptable in sensory test scores over 12 wk of storage at −0.3 °C, despite total viable count (TVC) and lactic acid bacteria (LAB) levels increasing to 8 log₁₀ CFU/cm² for VAC lamb and 4–6 log₁₀ CFU/cm² for MAP lamb. Similar to the sensory results, there were no significant differences in microbial communities between BI and BO product. However, types of bacteria were different between VAC and MAP packaging. Specifically, while VAC shoulder became dominated by Carnobacterium spp. in the middle of the storage period, the MAP shoulder microbial population remained similar from the start until later storage times.     

Interpersonal goals and change in anxiety and dysphoria in first-semester college students.

Two longitudinal studies examined the associations between interpersonal goals (i.e., self-image and compassionate goals) and anxiety and dysphoria (i.e., distress). In Study 1, 199 college freshmen (122 women, 77 men) completed 12 surveys over 12 weeks. Compassionate goals predicted decreased distress, and self-image goals predicted increased distress from pretest to posttest when distress was assessed as anxiety, dysphoria, or a composite, and when the goals were worded as approach goals, avoidance goals, or a composite. In Study 2, 115 first-semester roommate pairs (86 female and 29 male pairs) completed 12 surveys over 12 weeks. Compassionate and self-image goals predicted distress in same-week, lagged-week, and pretest-to-posttest analyses; effects of compassionate goals remained significant when the authors controlled for several known risk factors. Having clear goals consistently explained the association between compassionate goals but not self-image goals and distress. Results supported a path model in which compassionate goals predict increased support given to roommates, which predicts decreased distress. Results also supported a reciprocal association; chronic distress predicted decreased compassionate and increased self-image goals from pretest to posttest, and weekly distress predicted decreased compassionate goals the subsequent week. The results suggest that compassionate goals contribute to decreased distress because they provide meaning and increase support given to others. Distress, in turn, predicts change in goals, creating the potential for upward and downward spirals of goals and distress. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effect of plasma treatment on the surface properties of polydimethylsiloxane

In this work, the analysis of the plasma modification of polydimethylsiloxane (PDMS) substrates was conducted. The influence of the modification onto the changes occurred within the geometrical structure and chemical composition of the surface was analyzed. Due to the study of the atomic force microscopy, it was possible to determine the relationships between the surface development and applied process parameters, defining the conditions facilitating to obtain isotropic or anisotropic orientations of wrinkles. A precise analysis of the chemical composition of the surface, executed before and after the modification processes, enabled to define the changes in their elementary composition. Moreover, the modification influence onto the changes of the contact angle and the surface free energy were also analyzed. So, based on the research it was stated that the increase of value of the surface free energy is proportional to the sp³ phase contents in the DLC coatings. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41635.     

Measuring errors and violations on the road: A bifactor modeling approach to the Driver Behavior Questionnaire

The Driver Behavior Questionnaire (DBQ) is a self-report measure of driving behavior that has been widely used over more than 20 years. Despite this wealth of evidence a number of questions remain, including understanding the correlation between its violations and errors sub-components, identifying how these components are related to crash involvement, and testing whether a DBQ based on a reduced number of items can be effective. We address these issues using a bifactor modeling approach to data drawn from the UK Cohort II longitudinal study of novice drivers. This dataset provides observations on 12,012 drivers with DBQ data collected at .5, 1, 2 and 3 years after passing their test. A bifactor model, including a general factor onto which all items loaded, and specific factors for ordinary violations, aggressive violations, slips and errors fitted the data better than correlated factors and second-order factor structures. A model based on only 12 items replicated this structure and produced factor scores that were highly correlated with the full model. The ordinary violations and general factor were significant independent predictors of crash involvement at 6 months after starting independent driving. The discussion considers the role of the general and specific factors in crash involvement.