Ferrets exclusively synthesize Neu5Ac and express naturally humanized influenza A virus receptors

Mammals express the sialic acids ​N-acetylneuraminic acid (​Neu5Ac) and ​N-glycolylneuraminic acid (​Neu5Gc) on cell surfaces, where they act as receptors for pathogens, including influenza A virus (IAV). ​Neu5Gc is synthesized from ​Neu5Ac by the enzyme cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH). In humans, this enzyme is inactive and only ​Neu5Ac is produced. Ferrets are susceptible to human-adapted IAV strains and have been the dominant animal model for IAV studies. Here we show that ferrets, like humans, do not synthesize ​Neu5Gc. Genomic analysis reveals an ancient, nine-exon deletion in the ferret CMAH gene that is shared by the Pinnipedia and Musteloidia members of the Carnivora. Interactions between two human strains of IAV with the sialyllactose receptor (sialic acid—α2,6Gal) confirm that the type of terminal sialic acid contributes significantly to IAV receptor specificity. Our results indicate that exclusive expression of ​Neu5Ac contributes to the susceptibility of ferrets to human-adapted IAV strains

Cardiac expression of the cystic fibrosis transmembrane conductance regulator involves novel Exon 1 usage to produce a unique amino-terminal protein

Cystic fibrosis is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which encodes a chloride channel present in many cells. In cardiomyocytes, we report that multiple exon 1 usage and alternative splicing produces four CFTR transcripts, with different 5'-untranslated regions, CFTRTRAD-139, CFTR-1C/-1A, CFTR-1C, and CFTR-1B. CFTR transcripts containing the novel upstream exons (exons -1C, -1B, and -1A) represent more than 90% of cardiac expressed CFTR mRNA. Regulation of cardiac CFTR expression, in response to developmental and pathological stimuli, is exclusively due to the modulation of CFTR-1C and CFTR-1C/-1A expression. Upstream open reading frames have been identified in the 5'-untranslated regions of all CFTR transcripts that, in conjunction with adjacent stem-loop structures, modulate the efficiency of translation initiation at the AUG codon of the main CFTR coding region in CFTRTRAD-139 and CFTR-1C/-1A transcripts. Exon(-1A), only present in CFTR-1C/-1A transcripts, encodes an AUG codon that is in-frame with the main CFTR open reading frame, the efficient translation of which produces a novel CFTR protein isoform with a curtailed amino terminus. As the expression of this CFTR transcript parallels the spatial and temporal distribution of the cAMP-activated whole-cell current density in normal and diseased hearts, we suggest that CFTR-1C/-1A provides the molecular basis for the cardiac cAMP-activated chloride channel. Our findings provide further insight into the complex nature of in vivo CFTR expression, to which multiple mRNA transcripts, protein isoforms, and post-transcriptional regulatory mechanisms are now added

Die Weltwirtschaft im Wandel: Symposium 1975

Seit einigen Jahren verstärkt sich der Eindruck, daß die Weltwirtschaft in einem raschen Wandel begriffen ist. Die Entwicklungsländer fordern eine neue Weltwirtschaftsordnung. Auf die weltweite Hochkonjunktur ist eine scharfe und recht lange anhaltende Rezession gefolgt; aber noch immer dominieren inflationäre Tendenzen. Den Konjunktureinbruch überlagert hat die Vervielfachung des Ölpreises Ende 1973. Sie wurde von vielen Beobachtern als Beginn einer Phase mit zunehmendem Rohstoffmangel und nie gekannten Ungleichgewichten in den Zahlungsbilanzen gedeutet. Von den vielfach geäußerten Befürchtungen haben sich manche als übertrieben erwiesen; denn viele Schwierigkeiten konnte der Markt viel besser als erwartet und ziemlich geräuschlos bewältigen. Gleichwohl erscheint es geraten, die alten und die neuen Probleme der Weltwirtschaft ständig zu beobachten. Vom 23. -27. Juni, traf sich in Kiel eine Gruppe unabhängiger Ökonomen aus Japan, Nordamerika und der Europäischen Gemeinschaft in der Absicht, die Situation der Weltwirtschaft zu analysieren und Vorschläge für wirtschaftspolitische Maßnahmen auf nationaler und internationaler Ebene auszuarbeiten. Das Institut für Weltwirtschaft war Gastgeber. Das Kieler Symposium ist ein Glied in einer Reihe trilateraler Konferenzen über weltwirtschaftliche Fragen, die von der Brookings Institution in Washington, dem Japan Economic Research Center in Tokio und dem Institut für Weltwirtschaft gemeinsam veranstaltet werden. Die Ergebnisse werden jeweils in englischer, japanischer und deutscher Sprache veröffentlicht; frühere Berichte in der deutschen Fassung sind als Kieler Diskussionsbeiträge Nr. 31 "Welthandel und Strukturanpassung in den Industrieländern" und Nr. 36 "Weltrohstoffversorgung: Konflikt oder Kooperation?" erschienen

Die Weltwirtschaft im Wandel: Symposium 1975

Seit einigen Jahren verstärkt sich der Eindruck, daß die Weltwirtschaft in einem raschen Wandel begriffen ist. Die Entwicklungsländer fordern eine neue Weltwirtschaftsordnung. Auf die weltweite Hochkonjunktur ist eine scharfe und recht lange anhaltende Rezession gefolgt; aber noch immer dominieren inflationäre Tendenzen. Den Konjunktureinbruch überlagert hat die Vervielfachung des Ölpreises Ende 1973. Sie wurde von vielen Beobachtern als Beginn einer Phase mit zunehmendem Rohstoffmangel und nie gekannten Ungleichgewichten in den Zahlungsbilanzen gedeutet. Von den vielfach geäußerten Befürchtungen haben sich manche als übertrieben erwiesen; denn viele Schwierigkeiten konnte der Markt viel besser als erwartet und ziemlich geräuschlos bewältigen. Gleichwohl erscheint es geraten, die alten und die neuen Probleme der Weltwirtschaft ständig zu beobachten. Vom 23. -27. Juni, traf sich in Kiel eine Gruppe unabhängiger Ökonomen aus Japan, Nordamerika und der Europäischen Gemeinschaft in der Absicht, die Situation der Weltwirtschaft zu analysieren und Vorschläge für wirtschaftspolitische Maßnahmen auf nationaler und internationaler Ebene auszuarbeiten. Das Institut für Weltwirtschaft war Gastgeber. Das Kieler Symposium ist ein Glied in einer Reihe trilateraler Konferenzen über weltwirtschaftliche Fragen, die von der Brookings Institution in Washington, dem Japan Economic Research Center in Tokio und dem Institut für Weltwirtschaft gemeinsam veranstaltet werden. Die Ergebnisse werden jeweils in englischer, japanischer und deutscher Sprache veröffentlicht; frühere Berichte in der deutschen Fassung sind als Kieler Diskussionsbeiträge Nr. 31 "Welthandel und Strukturanpassung in den Industrieländern" und Nr. 36 "Weltrohstoffversorgung: Konflikt oder Kooperation?" erschienen. --

An insulator element 3′ to the CFTR gene binds CTCF and reveals an active chromatin hub in primary cells

Regulation of expression of the CFTR gene is poorly understood. Elements within the basal promoter of the gene do not fully explain CFTR expression patterns, suggesting that cis-regulatory elements are located elsewhere, either within the locus or in adjacent chromatin. We previously mapped DNase I hypersensitive sites (DHS) in 400 kb spanning the CFTR locus including a cluster of sites close to the 3′-end of the gene. Here we focus on a DHS at +6.8 kb from the CFTR translation end-point to evaluate its potential role in regulating expression of the gene. This DHS, which encompasses a consensus CTCF-binding site, was evident in primary human epididymis cells that express abundant CFTR mRNA. We show by DNase I footprinting and electophoretic mobility shift assays that the cis-regulatory element within this DHS binds CTCF in vitro. We further demonstrate that the element functions as an enhancer blocker in a well-established in vivo assay, and by using chromatin immunoprecipitation that it recruits CTCF in vivo. Moreover, we reveal that in primary epididymis cells, the +6.8 kb DHS interacts closely with the CFTR promoter, suggesting that the CFTR locus exists in a looped conformation, characteristic of an active chromatin hub

In vitro and in vivo MMP gene expression localisation by In Situ-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts

BACKGROUND: Members of the matrix metalloproteinase (MMP) family of proteases are required for the degradation of the basement membrane and extracellular matrix in both normal and pathological conditions. In vitro, MT1-MMP (MMP-14, membrane type-1-MMP) expression is higher in more invasive human breast cancer (HBC) cell lines, whilst in vivo its expression has been associated with the stroma surrounding breast tumours. MMP-1 (interstitial collagenase) has been associated with MDA-MB-231 invasion in vitro, while MMP-3 (stromelysin-1) has been localised around invasive cells of breast tumours in vivo. As MMPs are not stored intracellularly, the ability to localise their expression to their cells of origin is difficult. METHODS: We utilised the unique in situ-reverse transcription-polymerase chain reaction (IS-RT-PCR) methodology to localise the in vitro and in vivo gene expression of MT1-MMP, MMP-1 and MMP-3 in human breast cancer. In vitro, MMP induction was examined in the MDA-MB-231 and MCF-7 HBC cell lines following exposure to Concanavalin A (Con A). In vivo, we examined their expression in archival paraffin embedded xenografts derived from a range of HBC cell lines of varied invasive and metastatic potential. Mouse xenografts are heterogenous, containing neoplastic human parenchyma with mouse stroma and vasculature and provide a reproducible in vivo model system correlated to the human disease state. RESULTS: In vitro, exposure to Con A increased MT1-MMP gene expression in MDA-MB-231 cells and decreased MT1-MMP gene expression in MCF-7 cells. MMP-1 and MMP-3 gene expression remained unchanged in both cell lines. In vivo, stromal cells recruited into each xenograft demonstrated differences in localised levels of MMP gene expression. Specifically, MDA-MB-231, MDA-MB-435 and Hs578T HBC cell lines are able to influence MMP gene expression in the surrounding stroma. CONCLUSION: We have demonstrated the applicability and sensitivity of IS-RT-PCR for the examination of MMP gene expression both in vitro and in vivo. Induction of MMP gene expression in both the epithelial tumour cells and surrounding stromal cells is associated with increased metastatic potential. Our data demonstrate the contribution of the stroma to epithelial MMP gene expression, and highlight the complexity of the role of MMPs in the stromal-epithelial interactions within breast carcinoma

cDNA Immunization of Mice with Human Thyroglobulin Generates Both Humoral and T Cell Responses: A Novel Model of Thyroid Autoimmunity

Thyroglobulin (Tg) represents one of the largest known self-antigens involved in autoimmunity. Numerous studies have implicated it in triggering and perpetuating the autoimmune response in autoimmune thyroid diseases (AITD). Indeed, traditional models of autoimmune thyroid disease, experimental autoimmune thyroiditis (EAT), are generated by immunizing mice with thyroglobulin protein in conjunction with an adjuvant, or by high repeated doses of Tg alone, without adjuvant. These extant models are limited in their experimental flexibility, i.e. the ability to make modifications to the Tg used in immunizations. In this study, we have immunized mice with a plasmid cDNA encoding the full-length human Tg (hTG) protein, in order to generate a model of Hashimoto's thyroiditis which is closer to the human disease and does not require adjuvants to breakdown tolerance. Human thyroglobulin cDNA was injected and subsequently electroporated into skeletal muscle using a square wave generator. Following hTg cDNA immunizations, the mice developed both B and T cell responses to Tg, albeit with no evidence of lymphocytic infiltration of the thyroid. This novel model will afford investigators the means to test various hypotheses which were unavailable with the previous EAT models, specifically the effects of hTg sequence variations on the induction of thyroiditis

ATP-binding cassette (ABC) transporters in normal and pathological lung

ATP-binding cassette (ABC) transporters are a family of transmembrane proteins that can transport a wide variety of substrates across biological membranes in an energy-dependent manner. Many ABC transporters such as P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1) and breast cancer resistance protein (BCRP) are highly expressed in bronchial epithelium. This review aims to give new insights in the possible functions of ABC molecules in the lung in view of their expression in different cell types. Furthermore, their role in protection against noxious compounds, e.g. air pollutants and cigarette smoke components, will be discussed as well as the (mal)function in normal and pathological lung. Several pulmonary drugs are substrates for ABC transporters and therefore, the delivery of these drugs to the site of action may be highly dependent on the presence and activity of many ABC transporters in several cell types. Three ABC transporters are known to play an important role in lung functioning. Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene can cause cystic fibrosis, and mutations in ABCA1 and ABCA3 are responsible for respectively Tangier disease and fatal surfactant deficiency. The role of altered function of ABC transporters in highly prevalent pulmonary diseases such as asthma or chronic obstructive pulmonary disease (COPD) have hardly been investigated so far. We especially focused on polymorphisms, knock-out mice models and in vitro results of pulmonary research. Insight in the function of ABC transporters in the lung may open new ways to facilitate treatment of lung diseases