Standardizing Umbilical Cord Mesenchymal Stromal Cells for Translation to Clinical Use: Selection of GMP-Compliant Medium and a Simplified Isolation Method

Citation: Smith, J. R., Pfeifer, K., Petry, F., Powell, N., Delzeit, J., & Weiss, M. L. (2016). Standardizing Umbilical Cord Mesenchymal Stromal Cells for Translation to Clinical Use: Selection of GMP-Compliant Medium and a Simplified Isolation Method. Stem Cells International, 14. doi:10.1155/2016/6810980Umbilical cord derived mesenchymal stromal cells (UC-MSCs) are a focus for clinical translation but standardized methods for isolation and expansion are lacking. Previously we published isolation and expansion methods for UC-MSCs which presented challenges when considering good manufacturing practices (GMP) for clinical translation. Here, a new and more standardized method for isolation and expansion of UC-MSCs is described. The new method eliminates dissection of blood vessels and uses a closed-vessel dissociation following enzymatic digestion which reduces contamination risk and manipulation time. The new method produced >10 times more cells per cm of UC than our previous method. When biographical variables were compared, more UC-MSCs per gram were isolated after vaginal birth compared to Caesarian-section births, an unexpected result. UC-MSCs were expanded in medium enriched with 2%, 5%, or 10% pooled human platelet lysate (HPL) eliminating the xenogeneic serum components. When the HPL concentrations were compared, media supplemented with 10% HPL had the highest growth rate, smallest cells, and the most viable cells at passage. UC-MSCs grown in 10% HPL had surface marker expression typical of MSCs, high colony forming efficiency, and could undergo trilineage differentiation. The new protocol standardizes manufacturing of UC-MSCs and enables clinical translation

[Special Issue on SEA Demographics] Featured Article: Cambodian, Hmong, Lao and Vietnamese-Americans in the 2005 American Community Survey

The figures included in this short article are from the 2005 American Community Survey (ACS) released by the U.S. Census Bureau in late 2006. The 2005 ACS data set involves estimates based on surveys distributed to only a subset of the U.S. population and is thus problematic in some respects. This concise article is intended to provide basic 2005 demographic, educational and socioeconomic data related to Cambodian, Hmong, Lao and Vietnamese in the United States. It is not intended as a comprehensive explanatory research paper of factors underlying contemporary demographic, educational, and socioeconomic trends in these four ethnic communities. These topics should ideally be the focus of additional quantitative and qualitative research. Most of the figures used in this article are from the Cambodian alone or in any combination, Hmong alone or in any combination, Lao alone or in any combination, and Vietnamese alone or in any combination population profiles including in the 2005 ACS

Book Review: Nguyen, N. H. C. (2016). South Vietnamese Soldiers: Memories of the Vietnam War and After. Santa Barbara, CA: Praeger. 289 pp. ISBN: 978-1-4408-3241-3

Book review by Mark Pfeifer: Nguyen, N. H. C. (2016). South Vietnamese Soldiers: Memories of the Vietnam War and After. Santa Barbara, CA: Praeger. This work consists of oral histories of Vietnamese residing in Australia who served with the Republic of Vietnam Armed Forces (RVNAF) in the Vietnam War era

Wide-Scale Analysis of Human Functional Transcription Factor Binding Reveals a Strong Bias towards the Transcription Start Site

We introduce a novel method to screen the promoters of a set of genes with shared biological function, against a precompiled library of motifs, and find those motifs which are statistically over-represented in the gene set. The gene sets were obtained from the functional Gene Ontology (GO) classification; for each set and motif we optimized the sequence similarity score threshold, independently for every location window (measured with respect to the TSS), taking into account the location dependent nucleotide heterogeneity along the promoters of the target genes. We performed a high throughput analysis, searching the promoters (from 200bp downstream to 1000bp upstream the TSS), of more than 8000 human and 23,000 mouse genes, for 134 functional Gene Ontology classes and for 412 known DNA motifs. When combined with binding site and location conservation between human and mouse, the method identifies with high probability functional binding sites that regulate groups of biologically related genes. We found many location-sensitive functional binding events and showed that they clustered close to the TSS. Our method and findings were put to several experimental tests. By allowing a "flexible" threshold and combining our functional class and location specific search method with conservation between human and mouse, we are able to identify reliably functional TF binding sites. This is an essential step towards constructing regulatory networks and elucidating the design principles that govern transcriptional regulation of expression. The promoter region proximal to the TSS appears to be of central importance for regulation of transcription in human and mouse, just as it is in bacteria and yeast.Comment: 31 pages, including Supplementary Information and figure

A hybrid approach to forecasting wind power using Artificial Neural Networks and Numeric Weather Prediction

Thesis (M.S.)--Wichita State University, College of Engineering, Dept. of Electrical Engineering and Computer Science.A methodology to forecast wind power production 24 hours ahead is developed using a hybrid approach of an artificial neural network (ANN) and numerical weather prediction (NWP). The methodology is simple and designed to be applicable to any wind farm on the globe, using publicly available NWP data and basic historical power production data from wind farm. Notably, no historical wind data from on-farm sensors is required as the 0 hour forecast data is used to train the ANN. The results are encouraging, with a root-mean-square-error of 0.2267 for a 24 hour ahead forecast, corresponding to a forecast error standard deviation of 0.23 per unit

SSB1/SSB2 proteins safeguard B-cell development by protecting the genomes of B-cell precursors

Induction of programmed DNA damage and its recognition and repair are fundamental for B cell development. The ssDNA-binding protein SSB1 has been described in human cells as essential for the recognition and repair of DNA damage. To study its relevance for B cells, we recently developed Ssb1−/− and conditional Ssb1−/− mice. Although SSB1 loss did not affect B cell development, Ssb1−/− cells exhibited compensatory expression of its homolog SSB2. We have now generated Ssb2−/− mice and show in this study that SSB2 is also dispensable for B cell development and DNA damage response activation. In contrast to the single loss of Ssb1 or Ssb2, however, combined SSB1/2 deficiency caused a defect in early B cell development. We relate this to the sensitivity of B cell precursors as mature B cells largely tolerated their loss. Toxicity of combined genetic SSB1/2 loss can be rescued by ectopic expression of either SSB1 or SSB2, mimicked by expression of SSB1 ssDNA-binding mutants, and attenuated by BCL2-mediated suppression of apoptosis. SSB1/2 loss in B cell precursors further caused increased exposure of ssDNA associated with disruption of genome fragile sites, inefficient cell cycle progression, and increased DNA damage if apoptosis is suppressed. As such, our results establish SSB1/2 as safeguards of B cell development and unveil their differential requirement in immature and mature B lymphocytes

Medienpraktiken: situieren, erforschen, reflektieren; eine Einleitung

Mit dem Fokus auf Medienpraktiken bündelt dieses Heft aktuelle Positionen zur empirischen Erforschung von Medien. Die Beiträge gehen davon aus, dass Medien erst durch ihren Gebrauch zu Medien werden. Medienpraktiken zu erforschen, bedeutet jedoch nicht nur herauszufinden, was Menschen mit Medien tun, sondern auch was Medien mit Menschen machen. Diese für die Medienpraktikenforschung zentrale Einsicht lösen die interdisziplinären Beiträge des Bandes ein, indem sie aus den jeweiligen Positionen und Konstellationen verdeutlichen, wie Medien und Praktiken sich gegenseitig bedingen. Medienpraktikenforschung erfordert erstens, medienpraktische Phänomene in einem hohen Detailgrad zu fassen, um die Relation der beteiligten menschlichen und medialen Akteure zueinander in situ und in actu nachzuvollziehen. Erst durch die analytische Durchdringung dieser situativen Vollzugsmomente lässt sich zweitens der Status von Medien klären: was durch Praktiken zu einem Medium wird und wie die Praktiken unter Berücksichtigung der an ihnen konstitutiv beteiligten Medien beschaffen sind. Dadurch lassen sich ebenso übersituative Bezüge zur Praxis herstellen, durch die die Praktiken zur situativen Entfaltung kommen. Drittens muss dabei berücksichtigt werden, inwiefern die eigenen Medienpraktiken der Erforschung in ihren jeweiligen situativen Stadien die (Analyse der) Medienpraktik zurichten. Die Beiträge dieses Bandes lösen diese Forderungen in unterschiedlicher Gewichtung ein. Sie befassen sich aus medienethnologischer, kultursoziologischer, literaturwissenschaftlicher, historischer, soziologischer und medienwissenschaftlicher Perspektive damit, was jeweils als situierte Medienpraktik verstanden werden kann. Gemeinsam ist damit allen Beiträgen, dass sie erst aus ihren jeweiligen Untersuchungen und Perspektiven heraus bestimmen, was genau als Medienpraktik und Medien, die in ihnen zum Tragen kommen, gefasst werden kann

Accelerating root system phenotyping of seedlings through a computer-assisted processing pipeline

Background: There are numerous systems and techniques to measure the growth of plant roots. However, phenotyping large numbers of plant roots for breeding and genetic analyses remains challenging. One major difficulty is to achieve high throughput and resolution at a reasonable cost per plant sample. Here we describe a cost-effective root phenotyping pipeline, on which we perform time and accuracy benchmarking to identify bottlenecks in such pipelines and strategies for their acceleration. Results: Our root phenotyping pipeline was assembled with custom software and low cost material and equipment. Results show that sample preparation and handling of samples during screening are the most time consuming task in root phenotyping. Algorithms can be used to speed up the extraction of root traits from image data, but when applied to large numbers of images, there is a trade-off between time of processing the data and errors contained in the database. Conclusions: Scaling-up root phenotyping to large numbers of genotypes will require not only automation of sample preparation and sample handling, but also efficient algorithms for error detection for more reliable replacement of manual interventions