Immunological and Physiological Differences Between Layer- and Broiler Chickens after Concurrent Intratracheal Administration of Lipopolysaccharide and Human Serum Albumin

Layers and broilers were concurrently intratracheally challenged with 0.5 mg Lipopolysaccharide (LPS) and 0.1 mg Human Serum Albumin (HuSA) at 3 weeks of age. Specific total and isotype-specific (IgM, IgG, IgA) Antibody (Ab) responses to HuSA during 3 weeks following immunization, cellular in vitro mitogen responses to Concanavalin A (Con A) and specific cellular responses in vitro to different dosages of HuSA, blood serotonin (5-HT) levels, plasma Corticosterone (CORT) levels at 6 weeks of age and ex vivo nitric oxide (NO) production in the presence of LPS, respectively, were measured in all birds. Higher in vitro cellular responses to HuSA, but not Con A, were found in the broilers than in the layers. Also higher total, IgM and IgG antibody responses to HuSA were found in the broilers. Higher ex vivo NO production was found in the layers. A heavier spleen weight was found in the broilers, but relative spleen weight was higher in the layers. The broilers grew much heavier and also maintained a higher growth during the first 24 and 48 h after i.t. challenge with LPS and HuSA. No breed effect was found for body temperature responses after i.t. challenge. Blood 5-HT levels and plasma CORT levels were significantly higher in the layers. Number and type of significant correlations between 5-HT levels, cachectin response to LPS, antibody levels and cellular immunity differed between breeds. Our data suggest comparable immune responses to i.t. HuSA challenge in broilers and layers of similar age and confirm the earlier reported higher humoral immune response in broilers. On the other hand, the cachectin response to LPS differed between broilers and layers. Our results do not confirm the earlier reported higher cellular immune response of layers. Different significant relationships between physiological parameters in broilers and layers were found. Our results suggest that selection for enhanced growth does not necessarily affect specific immune competence of poultr

CFAP300 mutation causing primary ciliary dyskinesia in Finland

Primary ciliary dyskinesia (PCD) is a rare genetic condition characterized by chronic respiratory tract infections and in some cases laterality defects and infertility. The symptoms of PCD are caused by malfunction of motile cilia, hair-like organelles protruding out of the cell that are responsible for removal of mucus from the airways and organizing internal organ positioning during embryonic development. PCD is caused by mutations in genes coding for structural or assembly proteins in motile cilia. Thus far mutations in over 50 genes have been identified and these variants explain around 70% of all known cases. Population specific genetics underlying PCD has been reported, thus highlighting the importance of characterizing gene variants in different populations for development of gene-based diagnostics. In this study, we identified a recurrent loss-of-function mutation c.198_200delinsCC in CFAP300 causing lack of the protein product. PCD patients homozygous for the identified CFAP300 mutation have immotile airway epithelial cilia associated with missing dynein arms in their ciliary axonemes. Furthermore, using super resolution microscopy we demonstrate that CFAP300 is transported along cilia in normal human airway epithelial cells suggesting a role for CFAP300 in dynein complex transport in addition to preassembly in the cytoplasm. Our results highlight the importance of CFAP300 in dynein arm assembly and improve diagnostics of PCD in Finland

Development of a synthetic gene network to modulate gene expression by mechanical forces

The majority of (mammalian) cells in our body are sensitive to mechanical forces, but little work has been done to develop assays to monitor mechanosensor activity. Furthermore, it is currently impossible to use mechanosensor activity to drive gene expression. To address these needs, we developed the first mammalian mechanosensitive synthetic gene network to monitor endothelial cell shear stress levels and directly modulate expression of an atheroprotective transcription factor by shear stress. The technique is highly modular, easily scalable and allows graded control of gene expression by mechanical stimuli in hard-to-transfect mammalian cells. We call this new approach mechanosyngenetics. To insert the gene network into a high proportion of cells, a hybrid transfection procedure was developed that involves electroporation, plasmids replication in mammalian cells, mammalian antibiotic selection, a second electroporation and gene network activation. This procedure takes 1 week and yielded over 60% of cells with a functional gene network. To test gene network functionality, we developed a flow setup that exposes cells to linearly increasing shear stress along the length of the flow channel floor. Activation of the gene network varied logarithmically as a function of shear stress magnitude

Alquimia, Ocultismo, Maçonaria: o ouro e o simbolismo hermético dos cadinhos (Séculos XVIII e XIX)

Este artigo apresenta a arqueologia das enigmáticas marcas impressas na base de cadinhos dos séculos XVIII e XIX recuperados nas escavações da Casa da Moeda do Rio de Janeiro, na década de 1980, e a explanação do seu significado simbólico à luz da alquimia, do ocultismo e da Maçonaria. Espraiando-se extraordinariamente mundo afora através de uma bem-sucedida estratégia de comunicação visual, a Maçonaria utilizou símbolos herméticos para a difusão de seus princípios nos mais diferentes suportes. Aparentemente estamos diante de um sinal de reconhecimento maçônico, o sinal exterior de uma organização oculta, só partilhado por iniciados e incompreensível para os demais, que contribuiu para difundir veladamente a doutrina maçônica por diferentes pontos do globo

Generating mutant renal cell lines using CRISPR technologies

Gene-editing using the CRISPR/Cas9 system is an extremely efficient approach for generating mutations within the genomic DNA of immortalised cell lines. This procedure begins with a straightforward cloning step to generate a single plasmid encoding the Cas9 enzyme as well as a synthetic guide RNA (sgRNA) which is selected to target specific sites within the genome. This plasmid is transfected into cells either alone, in order to generate random insertion-deletion alleles (‘indels’) at the desired locus via the non-homologous end-joining pathway, or in conjunction with a homology directed repair template oligonucleotide to generate a specific point mutation. Here we describe a procedure to perform gene-editing in IMCD3 and HEK293 cells and to subsequently isolate clonal cell lines carrying mutations of interest