Swiss Startup Framework: A Highly Effective Network Supporting the Generation of Emerging Biotech Businesses

For over 50 years, Switzerland has been one of the leading countries driving innovation in biotechnology and its industrial applications. Today, some 1,000 biotech companies form a tightly knit, cross-functional network ranging from research through to manufacturing. This network comprises R&D companies, contract research organizations, and highly specialized advisors and biotech investors. Together, they form an external innovation pool that complements the in-house R&D capacity of the large multi-national pharma companies. A highly effective startup framework, solid acceleration mechanisms, and innovative investors enable the emergence of a continuous flow of biotech startups that revitalize the industry with new technologies and products supporting drug development and diagnostics

East Asia’s new developmentalism: state capacity, climate change and low-carbon development

This paper argues that to understand the relevance of developmental states in East Asia and elsewhere, we need to focus on the changing development agenda in the early twenty-first century, especially how this connects with the global challenge of climate change and thereby sustainable, low-carbon development. It combines theories on state capacity and ecological modernisation to form the ‘new developmentalism’ concept. This is applied to study revitalised and refocused forms of state capacity aimed at realising the transformative economic objectives associated with sustainable development. New developmentalism helps us understand not only current state capacity practice in a climate challenged world but also how we have moved beyond original conceptions of developmental statism. It may be understood in the wider context of the sustainable development agenda and climate interventionism. As is argued, new developmentalism is most clearly evident in East Asia but can be applied in a wider geographic sense where strong forms of developmental state capacity are exercised towards meeting transformative sustainable development goals

Effect of siRNAs, TGF-β1 and/or PDGF-AB on HSC proliferation.

<p>(A-B) hTERT-HSCs (A) and primary HSCs (B) were transfected for 72 hours with siNrf2, siKeap1 or siCON. Cells were then detached and plated for EdU staining as described in Materials and methods section. The pictures were taken with 10X magnification by using confocal microscopy and nuclei were counted with Image J software. siCON: scrambled siRNA; siNrf2: Nrf2 knockdown; siKeap1: Keap1 knockdown. *, P ≤ 0.05; **, P ≤ 0.01; *** P ≤ 0.001 vs control or siCON. Values are expressed as rate of proliferation (mean ± SD); N = 4 independent experiment with 5 replicates each for hTERT-HSC, and N = 6 different batches with 5 replicates each. (C-D) Proliferation rate of transfected hTERT-HSC (C) and primary HSC (D) after exposure to 0.5–1 ng/mL TGF-β1 and/or 1–5 ng/mL PDGF-AB for 48 hours. EdU was added during the last 30 hours of the experiment. Values are expressed as percentage of proliferation over each control sample. *, P ≤ 0.05; **, P ≤ 0.01; *** P ≤ 0.001 vs Control (mean ± SD, N = 4 replicates for hTERT-HSC and N = 3 from different batches with 4 replicates each for primary HSC).</p

TGF-β1 suppresses mRNA expression of Nrf2 in HSCs.

<p>(A) hTERT-HSCs were exposed to 1–5 ng/mL TGF-β1 for 48 hours. mRNA was extracted using TRIzol conventional procedure and fold changes were calculated as 2^(-ΔΔCT) for each sample and control and expressed as mean fold change ± SD (N = 3). Beta-2-microglobulin (B2M) was used as reference gene for each sample. The results show a significant downregulation of Nrf2, Keap1 and Nqo1 after exposure to TGF-β1. (B) Primary HSCs were exposed to 1 ng/mL TGF-β1, 1 μM SB-525334 (SB52) and/or 5 ng/mL PDGF-AB for 48 hours. mRNA was extracted using TRIzol conventional procedure and fold changes were calculated as 2^(-ΔΔCT) for each sample and control and expressed as mean fold change ± SD (N = 3 different batches). Beta-2-microglobulin (B2M) was used as reference gene for each sample. The results show a significant downregulation of Nrf2, and Nqo1 following exposure to TGF-β1. SB52 induces upregulation of Nrf2 and Nqo1, efficiently inhibiting the TGF-β1 effect. (C-D) The protein levels of Nrf2, Nqo1 and actin beta were analysed by Western blot analysis after exposure to 0–10 ng/mL TGF-β1 in hTERT-HSC (C) and primary HSCs (D) for 24 and 48 hours. (E-F) The protein levels of Keap1 and Tubulin were analysed by Western blot analysis after exposure to 0–10 ng/mL TGF-β1 in hTERT-HSC (E) and primary HSCs (F) for 24 and 48 hours.</p

TaqMan probes used for the research.

<p>TaqMan probes used for the research.</p

Nrf2 intervention into the TGF-β1/Smad signalling pathway.

<p>Panel A, basal condition: HSCs express high levels of both Nrf2 and its target genes (e.g. Nqo1), thereby controlling reactive oxygen species (ROS) level. Nrf2 also inhibits Smad pathway by binding directly to Smad protein or through the action of phosphatases [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0201044#pone.0201044.ref041" target="_blank">41</a>]. In these conditions, the TGF-β1/Smad pathway has a low activity, resulting in low level of αSMA, collagens and TGF-β1. Thus, HSC cells exhibit a quiescent phenotype. Panel B, Nrf2 knockdown: we have found out that Nrf2 knockdown, with a consequent decrease of its target genes, induces stellate cells activation. Decrease in Nrf2 was in fact associated with an increase in the levels of Extracellular matrix (ECM) components as well as αSMA and TGF-β1. TGF-β1 further induces the expression of ATF3, which acts as functional repressor of Nrf2. We found out that this siNrf2-induced stellate cell activation may be regulated by the Smad inhibitors SB-431542 hydrate (SB43) and SB-525334 (SB52), confirming the role of Nrf2 in relation to the Smad pathway.</p

Molecular and in vitro Toxicology at the FHNW: FH - HES

In the recently established Molecular Toxicology laboratory at the Institute for Chemistry and Bioanalytics of the School of Life Sciences in the FHNW, we aim to develop and apply in vitro models for the investigation of toxic effects, with a primary focus on liver and kidney. In collaboration with other institutions, we are developing multicellular type 2D and 3D cell culture assays to be able to closely mimic relevant in vivo situations. In parallel we are broadening the choice of available endpoint analyses

Pro-fibrotic compounds induce stellate cell activation, ECM-remodelling and Nrf2 activation in a human 3D-multicellular model of liver fibrosis.

Currently most liver fibrosis research is performed in vivo, since suitable alternative in vitro systems which are able to recapitulate the cellular events leading to liver fibrosis are lacking. Here we aimed at generating a system containing cells representing the three key players of liver fibrosis (hepatocyte, Kupffer cells and stellate cells) and assess their response to pro-fibrotic compounds such as TGF-β1, methotrexate (MTX) and thioacetamide (TAA).Human cell lines representing hepatocytes (HepaRG), Kupffer cell (THP-1 macrophages) and stellate cells (hTERT-HSC) were co-cultured using the InSphero hanging drop technology to generate scaffold-free 3D microtissues, that were treated with pro-fibrotic compounds (TGF-β1, MTX, TAA) for up to 14 days. The response of the microtissues was evaluated by determining the expression of cytokines (TNF-α, TGF-β1 and IL6), the deposition and secretion of ECM proteins and induction of gene expression of fibrosis biomarkers (e.g. αSMA). Induction of Nrf2 and Keap1, as key player of defence mechanism, was also evaluated.We could demonstrate that the multicellular 3D microtissue cultures could be maintained in a non-activated status, based on the low expression levels of activation markers. Macrophages were activated by stimulation with LPS and hTERT-HSC showed activation by TGF-β1. In addition, MTX and TAA elicited a fibrotic phenotype, as assessed by gene-expression and protein-deposition of ECM proteins such as collagens and fibronectin. An involvement of the antioxidant pathway upon stimulation with pro-fibrotic compounds was also observed.Here, for the first time, we demonstrate the in vitro recapitulation of key molecular and cellular events leading to liver fibrosis: hepatocellular injury, antioxidant defence response, activation of Kupffer cells and activation of HSC leading to deposition of ECM

Methotrexate-Induced Liver Injury Is Associated with Oxidative Stress, Impaired Mitochondrial Respiration, and Endoplasmic Reticulum Stress In Vitro

Low-dose methotrexate (MTX) is a standard therapy for rheumatoid arthritis due to its low cost and efficacy. Despite these benefits, MTX has been reported to cause chronic drug-induced liver injury, namely liver fibrosis. The hallmark of liver fibrosis is excessive scarring of liver tissue, triggered by hepatocellular injury and subsequent activation of hepatic stellate cells (HSCs). However, little is known about the precise mechanisms through which MTX causes hepatocellular damage and activates HSCs. Here, we investigated the mechanisms leading to hepatocyte injury in HepaRG and used immortalized stellate cells (hTERT-HSC) to elucidate the mechanisms leading to HSC activation by exposing mono- and co-cultures of HepaRG and hTERT-HSC to MTX. The results showed that at least two mechanisms are involved in MTX-induced toxicity in HepaRG: (i) oxidative stress through depletion of glutathione (GSH) and (ii) impairment of cellular respiration in a GSH-independent manner. Furthermore, we measured increased levels of endoplasmic reticulum (ER) stress in activated HSC following MTX treatment. In conclusion, we established a human-relevant in vitro model to gain mechanistical insights into MTX-induced hepatotoxicity, linked oxidative stress in HepaRG to a GSH-dependent and -independent pathway, and hypothesize that not only oxidative stress in hepatocytes but also ER stress in HSCs contribute to MTX-induced activation of HSCs

Electrospun decellularized extracellular matrix scaffolds promote the regeneration of injured neurons

Traumatic injury to the spinal cord (SCI) causes the transection of neurons, formation of a lesion cavity, and remodeling of the microenvironment by excessive extracellular matrix (ECM) deposition and scar formation leading to a regeneration-prohibiting environment. Electrospun fiber scaffolds have been shown to simulate the ECM and increase neural alignment and neurite outgrowth contributing to a growth-permissive matrix. In this work, electrospun ECM-like fibers providing biochemical and topological cues are implemented into a scaffold to represent an oriented biomaterial suitable for the alignment and migration of neural cells in order to improve spinal cord regeneration. The successfully decellularized spinal cord ECM (dECM), with no visible cell nuclei and dsDNA content < 50 ng/mg tissue, showed preserved ECM components, such as glycosaminoglycans and collagens. Serving as the biomaterial for 3D printer-assisted electrospinning, highly aligned and randomly distributed dECM fiber scaffolds (< 1 µm fiber diameter) were fabricated. The scaffolds were cytocompatible and supported the viability of a human neural cell line (SH-SY5Y) for 14 days. Cells were selectively differentiated into neurons, as confirmed by immunolabeling of specific cell markers (ChAT, Tubulin ß), and followed the orientation given by the dECM scaffolds. After generating a lesion site on the cell-scaffold model, cell migration was observed and compared to reference poly-ε-caprolactone fiber scaffolds. The aligned dECM fiber scaffold promoted the fastest and most efficient lesion closure, indicating superior cell guiding capabilities of dECM-based scaffolds. The strategy of combining decellularized tissues with controlled deposition of fibers to optimize biochemical and topographical cues opens the way for clinically relevant central nervous system scaffolding solutions