Double conditional human embryonic kidney cell line based on FLP and ΦC31 mediated transgene integration

<p>Abstract</p> <p>Background</p> <p>FLP recombinase mediated integration into a pre-integrated FRT site is routinely used to generate highly reproducible stable transgenic cell lines. In this study, we broaden the system of site specific integration by introducing ΦC31 integrase mediated integration into attP sites.</p> <p>Results</p> <p>We generated a HEK293 host cell line with a single copy FRT as well as an attP site allowing site specific integration of two distinct transgenes. To achieve conditional control, we used the tetracycline and Shld1 inducible systems. By introducing fluorescent reporters we show that integration and induction of two transgenes are completely independent. We applied this new technique to investigate the effect of HNF4α on proliferation of HEK293 cells by introducing HNF4α into each integration site. We obtained in two independent cell lines highly reproducible results that prove the usefulness of this novel HEK-attP/FRT cell line.</p> <p>Conclusions</p> <p>In this study we have established and applied a HEK-attP/FRT cell line that allows site specific integration of two conditional transgenes using the FLP recombinase as well as the ΦC31 integrase.</p

Differential regulation of serum microRNA expression by HNF1β and HNF1α transcription factors.

We aimed to identify microRNAs (miRNAs) under transcriptional control of the HNF1β transcription factor, and investigate whether its effect manifests in serum.This article is freely available via Open Access. Click on the 'Additional Link' above to access the full-text via the publisher's site.Published (Open Access

An integrodifferential approach to modeling, control, state estimation and optimization for heat transfer systems

In this paper, control-oriented modeling approaches are presented for distributed parameter systems. These systems, which are in the focus of this contribution, are assumed to be described by suitable partial differential equations. They arise naturally during the modeling of dynamic heat transfer processes. The presented approaches aim at developing finite-dimensional system descriptions for the design of various open-loop, closed-loop, and optimal control strategies as well as state, disturbance, and parameter estimation techniques. Here, the modeling is based on the method of integrodifferential relations, which can be employed to determine accurate, finite-dimensional sets of state equations by using projection techniques. These lead to a finite element representation of the distributed parameter system. Where applicable, these finite element models are combined with finite volume representations to describe storage variables that are—with good accuracy—homogeneous over sufficiently large space domains. The advantage of this combination is keeping the computational complexity as low as possible. Under these prerequisites, real-time applicable control algorithms are derived and validated via simulation and experiment for a laboratory-scale heat transfer system at the Chair of Mechatronics at the University of Rostock. This benchmark system consists of a metallic rod that is equipped with a finite number of Peltier elements which are used either as distributed control inputs, allowing active cooling and heating, or as spatially distributed disturbance inputs

Tissue-specific transcription factor HNF4alpha inhibits cell proliferation and induces apoptosis in the pancreatic INS-1 beta-cell line

Hepatocyte nuclear factor 4alpha (HNF4alpha) is a tissue-specific transcription factor expressed in many cell types, including pancreatic beta-cells. Mutations in the HNF4alpha gene in humans give rise to maturity-onset diabetes of the young (MODY1) characterized by defective insulin secretion by beta-cells. To elucidate the mechanism underlying this disease, we introduced the splice form HNF4alpha2 or HNF4alpha8 into the rat beta-cell line INS-1. Upon tetracycline-induced expression, both HNF4alpha isoforms caused distinct changes in cell morphology and a massive loss of cell numbers that was correlated with reduced proliferation and induced apoptosis. This differential activity was reflected in oligonucleotide microarray analysis that identified more genes affected by HNF4alpha2 compared to HNF4alpha8, and suggests that both isoforms regulate largely the same set of genes, with HNF4alpha2 being a stronger transactivator. We verified the induction of selected transcripts by real-time RT-PCR, including KAI1 and AIF, both known to have apoptotic potential. By establishing cell lines with inducible expression of these target genes, we deduce that both factors are insufficient to induce apoptosis. We propose that the anti-proliferative and apoptotic properties of HNF4alpha may be an essential feature impaired in MODY1 and possibly also in type 2 diabetes

Are pit and fissure sealants needed in children with a higher caries risk?

The aim of this cross-sectional study was to analyse the preventive need of pit and fissure sealants (PFS) in a German population with a relatively high caries risk. The study involved 311 8- to 12-year-old children from the Ennepe-Ruhr District in North Rhine-Westphalia, Germany. Caries experience was scored according to WHO (1997) and ICDAS II criteria. PFS were assessed as intact or partially lost. The mean DFS values amounted to 0.5 for occlusal fissures, 0.2 for palatal/buccal pits and 0.3 for the remaining teeth. Non-cavitated caries lesions were recorded in average on 1.8 occlusal fissures and 1.5 palatal/buccal pits. Sealants were registered on 1.4 occlusal fissures and 0.4 palatal/buccal pits. The descriptive data and the adjusted Poisson regression models revealed that children with at least one fissure sealant are less likely to have decayed fissures or fissures with non-cavitated lesions on their permanent molars. Therefore, PFS are needed and indicated in caries-risk children