Einsatz von Menbuton und Dexamethason-21-isonicotinat zur Stoffwechsel- und Leistungsstabilisierung bei Milchkühen post partum

Ziel dieser Arbeit war es, unter Feldbedingungen zu untersuchen, ob eine einmalige Applikation von Dexamethason-21-isonicotinat, von Menbuton sowie eine kombinierte Anwendung beider Medikamente im frühen postpartalen Zeitraum bei Milchkühen eine stoffwechselstabilisierende Wirkung und einen Einfluss auf die Gesundheit, Fruchtbarkeit und Milchleistung haben

The potential impact of Wnt5a on differentiation and phenotype of dendritic cells found in renal cell carcinoma

Renal cell carcinoma (RCC) is an immunogenic tumour infiltrated by a high number of leucocytes. However, this infiltrate is not able to induce an efficient anti-tumoural response. Recent studies have attempted to further identify the composition of the immune-infiltrate in order to gain a deeper understanding of how RCC escapes from immune surveillance. Recently, a new dendritic cell (DC) subtype was discovered, co-expressing DC (CD209/DC-SIGN) and Mφ (CD14, CD163) markers. As this subtype was found to be enriched in RCC tissue it was called “enriched in renal cell carcinoma DC” (ercDC). Functional tests investigating cytokine expressions and the migration ability of this subtype revealed impaired functioning of ercDC compared to conventional DC (cDC). While being able to migrate towards fMLP, a chemoattractant for immature DC, maturated ercDC do not show directed migration towards CCL19, which is involved in the recirculation and homing of maturated DC. Furthermore, ercDC secrete higher levels of IL-10 and TNF-α, but lower levels of IL-12 upon LPS stimulation. In order to identify possible pathways responsible for differences between myeloid subtypes, transcriptomic profiling of monocytes, cDC, ercDC and Mφ was performed. Amongst others, regulation was found especially for the canonical as well as non-canonical Wntpathways. Following these findings, we investigated the role of Wnt-signalling in the differentiation and functioning of myeloid and especially dendritic cells. Striking was the high expression of Wnt5a, a typical non-canonical Wnt-ligand, in cDC in comparison to all other myeloid subtypes. Furthermore, we could show that Wnt5a is highly expressed in RCC and microarray analysis of RCC tissue samples revealed that Wnt-signalling, especially non-canonical Wnt-signalling, changes with tumour progression. Functional tests of DC showed, that the addition of Wnt5a during differentiation significantly increases IL-10 and IL-12, but decreases TNF-α secretion. Surprisingly, Wnt5a also inhibited the migratory capacity of cDC, resulting in reduced motility and less directed migration towards CCL19. These results suggest a possible role of Wntsignalling in the differentiation and functioning of myeloid cells. Through its autocrine, paracrine and juxtacrine effect, Wnt5a present in the tumour milieu may influence the anti-tumoural immune response. Gaining further insight into the role of myeloid cells and Wnt-signalling in RCC could hence reveal new possible therapeutic targets.Das Nierenzellkarzinom (RCC, renal cell carcinoma) wird als immunogener Tumor von einer Vielzahl an Immunzellen infiltriert. Trotz dieses Immunzellinfiltrats, kommt es jedoch nicht zu einer effektiven Kontrolle des Tumorwachstums durch das Immunsystem. Um die Mechanismen der Immunevasion des Nierenzellkarzinoms besser zu verstehen, ist das Ziel neuerer Studien, dieses Immuninfiltrat besser zu charakterisieren. Kürzlich wurde hierbei ein neuer myeloischer Zelltyp entdeckt, welcher sowohl dendritische Zell (DC)- (CD209/DC-SIGN), als auch Makrophagen (Mφ)- Marker (CD14, CD163) exprimiert. Da dieser Zelltyp im RCC im Vergleich zu tumorfreien Nierenparenchym besonders angereichert ist, wurde er „enriched in renal cell carcinoma DC“ (ercDC) genannt. Bei unserer Untersuchung dieses Zelltyps zeigte sich interessanterweise, dass maturierte ercDC zwar zu fMLP, einem Chemoattractant unreifer DC, gerichtet migrieren, allerdings keine gerichtete Migration zu CCL19 aufweisen. Im Vergleich zu konventionellen Dendritischen Zellen (cDC) sezernierten ercDC nach Stimulation mit LPS höhere Mengen IL-10 und TNF-α, aber geringere Mengen IL-12. Um mögliche relevante Unterschiede in den Signalwegen von Monozyten, cDC, ercDC und Mφ zu identifizieren, führten wir Transkriptomanalysen an Microarray-Daten dieser Zelltypen durch. Hierbei zeigte sich unter anderem eine differenzielle Expression in dem „canonical“ und den „non-canonical“ Wnt-Signalwegen. Auffallend war die, im Vergleich zu den anderen myeloischen Zelltypen, starke Expression von Wnt5a, einem typischen „non-canonical“ Wnt-Liganden, in cDC. Darüber hinaus konnten wir immunhistochemisch eine hohe Wnt5a Expression im Gewebe des Nierenzellkarzinoms nachweisen. Auch in der Microarray-Analyse von Nierenzellkarzinomproben zeigte sich eine starke differenzielle Expression der Wnt-Signalwege, mit evidenten Unterschieden abhängig vom Tumorstadium. Durch die Zugabe von Wnt5a während der Differenzierung von cDC konnte eine vermehrte Sekretion von IL-10 und IL-12, sowie eine Reduktion der TNF-α Sekretion erreicht werden. Interessanterweise führte Wnt5a Zugabe auch zu einer geringeren Mobilität und Direktionalität von cDC bei der Migration zu CCL19. Zusammenfassend weisen diese Ergebnisse auf eine Rolle der Wnt-Signalwege in der Differenzierung und Funktionalität von myeloischen Zellen hin. Wnt5a im Tumormilieu kann so durch autokirne, parakrine und juxtakrine Mechanismen die Immunantwort gegen den Tumor modulieren. Ein weitergehendes Verständnis über die Bedeutung des Wnt-Signalweges in den myeloischen Zellen des Nierenzellkarzinoms könnte so sogar mögliche neue Therapieansätze aufdecken

Reciprocal regulatory circuits coordinating EMT plasticity

Epithelial to mesenchymal transition (EMT) as well as its reversal process, mesenchymal to epithelial transition (MET), are essential and well-controlled cellular processes during embryonic development. Tightly controlled regulatory mechanisms guide an EMT/MET plasticity and enable cells to switch forth and back between different cell morphologies and functional capabilities to endow the necessity of tissue plasticity. However, aberrant and uncontrolled activation of these processes during malignant tumor progression promotes primary tumor cell invasion, cancer cell dissemination and metastatic outgrowth. In a recent study (Nat Commun; doi: 10.1038/s41467-017-01197-w), we have reported on the post-transcriptional control of normal and cancer-associated EMT by miRNAs and identified a novel, critical double-negative feedback regulation of the thus far unknown miRNA miR1199 and the key EMT transcription factor Zeb1

Governor’s proclamation

This proclamation from Governor Mark Sanford proclaims June 22, 2004, as Bill Shaw Day

Guidelines for defining assembly batches of engineer-toorder concrete prefabricated systems

Os potenciais benefícios do uso de sistemas construtivos pré-fabricados do tipo engineer-to-order (ETO) abrangem desde a redução de custos e duração do empreendimento até melhorias na qualidade do produto. Entretanto, o escopo de sistemas de planejamento e controle da produção neste contexto deve ser diferente de empreendimentos tradicionais, pois existe a necessidade de coordenar diferentes tipos de fluxos (projeto, fabricação, operações logísticas e montagem). Neste contexto, é necessária a estabilização desses fluxos e consequente diminuição do trabalho em progresso, o que é relacionado ao esforço de reduzir o tamanho do lote de produção. O objetivo do presente trabalho é propor um conjunto de diretrizes para a definição dos lotes de montagem para sistemas pré-fabricados de concreto do tipo ETO com apoio de BIM 4D, com base em conceitos e princípios da produção enxuta. Design Science Research foi a abordagem metodológica adotada, tendo como base um estudo empírico em uma obra de ampliação de um aeroporto. As principais contribuições do trabalho são um conjunto de categorias de decisão para a definição dos lotes de montagem, e o desenvolvimento de ferramentas para apoiar a tomada de decisões.The potential benefits of engineer-to-order (ETO) prefabricated building systems include reductions in project cost and duration, and improvements in product quality. However, the scope of production planning and control systems in that context must be different from traditional projects, since it is necessary to coordinate different types of flows (design, manufacturing, logistics operations and site assembly). In this context, it is necessary to stabilise those flows and consequently reduce work in progress, which is strongly related to the effort of reducing production batch size. The aim of this research work is to propose a set of guidelines for the definition of assembly batches for ETO prefabricated concrete system, with the support of 4D BIM, based on concepts and principles of the lean production philosophy. Design Science Research was the methodological approach adopted in this investigation, which was based on an empirical study carried out in an airport expansion project. The main contributions of this study are a set of decision categories for the definition of assembly batches, and the development of tools for supporting decision-making in this context

Endothelial cell-derived nidogen-1 inhibits migration of SK-BR-3 breast cancer cells

BACKGROUND: The tumour microenvironment is a critical regulator of malignant cancer progression. While endothelial cells have been widely studied in the context of tumour angiogenesis, their role as modulators of cancer cell invasion and migration is poorly understood. METHODS: We have investigated the influence of endothelial cells on the invasive and migratory behaviour of human cancer cells in vitro. RESULTS: Upon exposure to culture supernatants of endothelial cells, distinct cancer cells, such as SK-BR-3 cells, showed significantly increased invasion and cell migration concomitant with changes in cell morphology and gene expression reminiscent of an epithelial-mesenchymal transition (EMT). Interestingly, the pro-migratory effect on SK-BR-3 cells was significantly enhanced by supernatants obtained from subconfluent, proliferative endothelial cells rather than from confluent, quiescent endothelial cells. Systematically comparing the supernatants of subconfluent and confluent endothelial cells by quantitative MS proteomics revealed eight candidate proteins that were secreted at significantly higher levels by confluent endothelial cells representing potential inhibitors of cancer cell migration. Among these proteins, nidogen-1 was exclusively expressed in confluent endothelial cells and was found to be necessary and sufficient for the inhibition of SK-BR-3 cell migration. Indeed, SK-BR-3 cells exposed to nidogen-1-depleted endothelial supernatants showed increased promigratory STAT3 phosphorylation along with increased cell migration. This reflects the situation of enhanced SK-BR-3 migration upon stimulation with conditioned medium from subconfluent endothelial cells with inherent absence of nidogen-1 expression. CONCLUSION: The identification of nidogen-1 as an endothelial-derived inhibitor of migration of distinct cancer cell types reveals a novel mechanism of endothelial control over cancer progression

Mesenchymal Cancer Cell-Stroma Crosstalk Promotes Niche Activation, Epithelial Reversion, and Metastatic Colonization

SummaryDuring metastatic colonization, tumor cells must establish a favorable microenvironment or niche that will sustain their growth. However, both the temporal and molecular details of this process remain poorly understood. Here, we found that metastatic initiating cells (MICs) exhibit a high capacity for lung fibroblast activation as a result of Thrombospondin 2 (THBS2) expression. Importantly, inhibiting the mesenchymal phenotype of MICs by blocking the epithelial-to-mesenchymal transition (EMT)-associated kinase AXL reduces THBS2 secretion, niche-activating ability, and, consequently, metastatic competence. Subsequently, disseminated metastatic cells revert to an AXL-negative, more epithelial phenotype to proliferate and decrease the phosphorylation levels of TGF-β-dependent SMAD2-3 in favor of BMP/SMAD1-5 signaling. Remarkably, newly activated fibroblasts promote this transition. In summary, our data reveal a crosstalk between cancer cells and their microenvironment whereby the EMT status initially triggers and then is regulated by niche activation during metastatic colonization

Live slow-frozen human tumor tissues viable for 2D, 3D, ex vivo cultures and single-cell RNAseq

Biobanking of surplus human healthy and disease-derived tissues is essential for diagnostics and translational research. An enormous amount of formalin-fixed and paraffin-embedded (FFPE), Tissue-Tek OCT embedded or snap-frozen tissues are preserved in many biobanks worldwide and have been the basis of translational studies. However, their usage is limited to assays that do not require viable cells. The access to intact and viable human material is a prerequisite for translational validation of basic research, for novel therapeutic target discovery, and functional testing. Here we show that surplus tissues from multiple solid human cancers directly slow-frozen after resection can subsequently be used for different types of methods including the establishment of 2D, 3D, and ex vivo cultures as well as single-cell RNA sequencing with similar results when compared to freshly analyzed material

TGF-β uses a novel mode of receptor activation to phosphorylate SMAD1/5 and induce epithelial-to-mesenchymal transition

The best characterized signaling pathway downstream of transforming growth factor β (TGF-β) is through SMAD2 and SMAD3. However, TGF-β also induces phosphorylation of SMAD1 and SMAD5, but the mechanism of this phosphorylation and its functional relevance is not known. Here, we show that TGF-β-induced SMAD1/5 phosphorylation requires members of two classes of type I receptor, TGFBR1 and ACVR1, and establish a new paradigm for receptor activation where TGFBR1 phosphorylates and activates ACVR1, which phosphorylates SMAD1/5. We demonstrate the biological significance of this pathway by showing that approximately a quarter of the TGF-β-induced transcriptome depends on SMAD1/5 signaling, with major early transcriptional targets being the ID genes. Finally, we show that TGF-β-induced epithelial-to-mesenchymal transition requires signaling via both the SMAD3 and SMAD1/5 pathways, with SMAD1/5 signaling being essential to induce ID1. Therefore, combinatorial signaling via both SMAD pathways is essential for the full TGF-β-induced transcriptional program and physiological responses.publishe

Transforming growth factor-, matrix metalloproteinases, and urokinase-type plasminogen activator interaction in the cancer epithelial to mesenchymal transition

Transforming growth factor- (TGF-) is a pleiotropic factor that acts as a tumor suppressor in the early stages, while it exerts tumor promoting activities in advanced stages of cancer development. One of the hallmarks of cancer progression is the capacity of cancer cells to migrate and invade surrounding tissues with subsequent metastasis to different organs. Matrix metalloproteinases (MMPs) together with urokinase-type plasminogen activator (uPA) and its receptor (uPAR), whose main original function described is the proteolytic degradation of the extracellular matrix, play key cellular roles in the enhancement of cell malignancy during cancer progression. TGF- tightly regulates the expression of several MMPs and uPA/uPAR in cancer cells, which in return can participate in TGF- activation, thus contributing to tumor malignancy. TGF- is one of the master factors in the induction of cancer-associated epithelial to mesenchymal transition (EMT), and recently both MMPs and uPA/uPAR have also been shown to be implicated in the cancer-associated EMT process. In this review, we analyze the main molecular mechanisms underlying MMPs and uPA/uPAR regulation by TGF-, as well as their mutual implication in the development of EMT in cancer cells. Developmental Dynamics 247:382-395, 2018