This protocol details the induction of a hemogenic program in mouse embryonic fibroblasts (MEFs) via overexpression of transcription factors (TFs). We first designed a reporter screen using MEFs from human CD34-tTA/TetO-H2BGFP (34/H2BGFP) double transgenic mice. CD34+ cells from these mice label H2B histones with GFP, and cease labeling upon addition of doxycycline (DOX). MEFS were transduced with

Previous attempts to either generate or expand hematopoietic stem cells (HSCs) in vitro have involved either ex vivo expansion of pre-existing patient or donor HSCs or de novo generation from pluripotent stem cells (PSCs), comprising both embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). iPSCs alleviated ESC ethical issues but attempts to generate functional mature hematopoie

Epigenetic reprogramming can be achieved in different ways, including nuclear transfer, cell fusion, or the expression of transcription factors (TFs). Combinatorial overexpression provides an opportunity to define the minimal core network of TFs that instructs specific cell fates. This approach has been employed to induce mouse and human pluripotency and differentiated cell types from cells that c

Many signals must be integrated to maintain self-renewal and pluripotency in embryonic stem cells (ESCs) and to enable induced pluripotent stem cell (iPSC) reprogramming. However, the exact molecular regulatory mechanisms remain elusive. To unravel the essential internal and external signals required for sustaining the ESC state, we conducted a short hairpin (sh) RNA screen of 104 ESC-associated p

The homeodomain transcription factor Nanog plays an important role in embryonic stem cell (ESC) self-renewal and is essential for acquiring ground-state pluripotency during reprogramming. Understanding how Nanog is transcriptionally regulated is important for further dissecting mechanisms of ESC pluripotency and somatic cell reprogramming. Here, we report that Nanog is subjected to a negative auto

In T lymphocytes, lipid rafts are preferred sites for signal transduction initiation and amplification. Many cell membrane receptors, such as the TCR, coreceptors, and accessory molecules associate within these microdomains upon cell activation. However, it is still unclear in most cases whether these receptors interact with rafts through lipid-based amino acid modifications or whether raft insert

We have recently reported that red blood cells (RBC) promote T cell growth and survival by inhibiting activation-induced T cell death. In the present study, we have examined parameters of oxidative stress and intracellular iron in activated T cells and correlated these data with the expression of ferritin, heme oxygenase-1 (HO-1), and the transferrin receptor CD71. T cells growing in the presence

The European Spallation Source (Lund, Sweden) is an under construction multi-disciplinary research facility to be based around a Linear Particle Accelerator which is to provide 2.86 ms long proton pulses at 2 GeV at a pulse repetition rate of 14 Hz, representing an average beam power of 5 MW. To accommodate the requirements of the proton linac, a large number of klystrons driven by power electroni

It is well-known that dyadic martingale transforms are a good model for Calderón–Zygmund singular integral operators. In this paper we extend some results on weighted norm inequalities to vector-valued functions. We prove that if W is an A2 matrix weight, then the weighted L2-norm of a Calderón–Zygmund operator with cancellation has the same dependence on the A2 characteristic of W as the weighted

An explicit expression, free from adjustable parameters, is derived for the effective pair interaction between charged colloidal spheres at high concentration in a medium containing an electrolyte. This is accomplished by first considering the electrostatic interaction between two infinite charged plates placed in a stack of identical plates. These act as a reservoir defining the chemical potentia

A heterogeneous distribution of crosslinker in micro-hydrogels (microgels) results in a non-uniform polymer density inside the particles. Identifying the morphology of the hydrogel backbone enables a bottom-up approach towards the structural and rheological properties of microgel systems. On a local level we use a Flory-Rehner inspired model that focuses on highly swollen networks, characterized b

The human cathelicidin peptide LL-37 has antimicrobial and anti-biofilm functions, but LL-37 may also damage the host by triggering inflammation and exerting a cytotoxic effect, thereby reducing host cell viability. Human plasma mitigates LL-37-induced host cell cytotoxicity but the underlying mechanisms are not completely understood. Apolipoprotein A-I (ApoA-I) is a plasma protein endowed with at

Background: There is accumulating evidence that digital breast tomosynthesis, referred to as 3D-mammography in this protocol, improves screen-detection measures compared to standard 2D-mammography in the context of population screening for breast cancer. However, the effect of 3D-mammography at follow-up of screened women is not yet known: it is unknown whether additional cancer detection from 3D-

Cardiovascular magnetic resonance imaging has become the gold standard for evaluating myocardial function, volumes, and scarring. Additionally, cardiovascular magnetic resonance imaging is unique in its comprehensive tissue characterization, including assessment of myocardial edema, myocardial siderosis, myocardial perfusion, and diffuse myocardial fibrosis. Cardiovascular magnetic resonance imagi

Recent studies have reported a very high frequency of noncoagulating milk in Swedish Red cows. The underlying factors are not fully understood. In this study, we explored rennet-induced coagulation properties and relative protein profiles in milk from native Swedish Mountain and Swedish Red Polled cows and compared them with a subset of noncoagulating (NC) and well-coagulating (WC) milk samples fr

A way to reduce net CO2 emission and circumvent the high particle emissions from compression ignition (CI) engines, while retaining high efficiency, is by utilizing lighter alcohols in the partially premixed combustion (PPC) process. Methanol and ethanol have a very short carbon chain, and form less soot/particulate matter (PM) during combustion compared to diesel fuel. This study analyzes and com

In the present study, the daily dose in terms of particle surface area received by citizens living in five cities in Western countries, characterized by different lifestyle, culture, climate and built-up environment, was evaluated and compared. For this purpose, the exposure to sub-micron particle concentration levels of the population living in Barcelona (Spain), Cassino (Italy), Guilford (United

We introduce successive cancellation (SC) decoding of product codes (PCs) with single parity-check (SPC) component codes. Recursive formulas are derived, which resemble the SC decoding algorithm of polar codes. We analyze the error probability of SPC-PCs over the binary erasure channel under SC decoding. A bridge with the analysis of PCs introduced by Elias in 1954 is also established. Furthermore

Novel experimental data of density for mixtures containing CO2 + ethanol, CO2 + ethyl lactate, CO2 + glycerol + ethanol, and CO2 + glycerol + ethyl lactate at low/moderate temperatures (308 and 323 K), pressures between 5 and 30 MPa, and high amounts of the organic solvent (molar fraction x = 0.5-0.9) are presented. The chosen organic solvents (ethanol, ethyl lactate, and glycerol) are green and a