Bone Marrow Transplantation In A Mouse Model Of Cooley’S Anemia

β-thalassemia is a group of inherited blood disorders that result in defects in β-globin chain production. Cooley’s anemia (CA), or β-thalassemia major, is the most severe form of the disease and occurs when an individual has mutations in both copies of the adult β-globin gene. Patients with CA fail to make adult hemoglobin, have ineffective erythropoiesis, suffer from severe anemia, and are transfusion dependent for life. Currently, allogeneic bone marrow transplantation (BMT) is the only cure; however, few patients have suitable donors for this procedure that carries a significant risk of morbidity and mortality. To develop new and safer alternatives to treatment of CA, a mouse model was developed to test novel therapies. As mice do not contain a fetal hemoglobin, a human γ-to-β globin switching cassette was knocked into the murine β-globin locus. When this cassette contained a hereditary persistence of fetal hemoglobin mutation in the γ-globin promoter and a non-functional human β0-globin gene, mice homozygous for the knockin were born alive, surviving solely on human fetal hemoglobin, and then demised as the γ-to-β globin switch was completed. This humanized CA mouse model yielded mice with a mean survival of two weeks. CA mice are rescued by chronic blood transfusions or cured by bone marrow transplantation. In the clinical setting, bone marrow transplantation is preceded by cytotoxic conditioning to make niche space for donor bone marrow cells to engraft. I hypothesized that by transplanting CA mice shortly after birth, cytotoxic conditioning could be avoided by exploiting the naivety of the newborn immune system. Strikingly, newborn CA mice could be rescued by BMT in the absence of cytoreductive conditioning. On the second day of life CA mice received a single antibody as conditioning and were transplanted with allogeneic cells on the following day. Transplanted mice were transfusion independent and showed physiological hematology and amelioration of disease related pathology. BMT in the absence of cytoreductive conditioning resulted in stable low-level donor hematopoietic chimerism that was capable of reconstituting the erythron. However, when donor bone marrow contained a major histocompatibility mismatch, donor cells were rejected and CA mice could not be rescued

Microorganisms Found in Field Specimens Of Diseased Corn Borer Larvae

Two hundred and eighty-six abnormal, field collected larvae of the European corn borer were examined for the presence of microorganisms which could have caused the abnormality or death. The most common microorganism found was a microsporidian, Perezia pyraustae Paillot. Next most numerous was a group of enterobacteria. Spore forming rods apparently of the genus Bacillus, and fungi belonging to the genera Beauvaria and Metarrhizium were also isolated. An examination of 100 apparently normal pupae revealed that 82 were hosts to Perezia pyraustae and two contained bacteria

Some Characteristics of Bacteria Isolated From Diseased Larvae of the European Corn Borer

A large proportion of the bacteria isolated from diseased corn borer larvae are pleomorphic, with rods, diplococci, filaments and various transition forms occurring in many cultures. Gram reaction is negative, although the coccoid forms show a tendency to retain gram positivity. In morphology and biochemical characteristics, these isolates resemble certain entomogenous bacteria described by earlier workers, but are similar also to a group of bacteria from human sources which have been designated as members of the tribe Mimeae DeBord. Further studies have been initiated to determine the pathogenicity and the proper taxonomic position of these cultures

Interactions between Type 1 Interferons and the Th17 Response in Tuberculosis: Lessons Learned from Autoimmune Diseases

textabstractThe classical paradigm of tuberculosis (TB) immunity, with a central protective role for Th1 responses and IFN-γ-stimulated cellular responses, has been challenged by unsatisfactory results of vaccine strategies aimed at enhancing Th1 immunity. Moreover, preclinical TB models have shown that increasing IFN-γ responses in the lungs is more damaging to the host than to the pathogen. Type 1 interferon signaling and altered Th17 responses have also been associated with active TB, but their functional roles in TB pathogenesis remain to be established. These two host responses have been studied in more detail in autoimmune diseases (AID) and show functional interactions that are of potential interest in TB immunity. In this review, we first identify the role of type 1 interferons and Th17 immunity in TB, followed by an overview of interactions between these responses observed in systemic AID. We discuss (i) the effects of GM-CSF-secreting Th17.1 cells and type 1 interferons on CCR2+ monocytes; (ii) convergence of IL-17 and type 1 interferon signaling on stimulating B-cell activating factor production and the central role of neutrophils in this process; and (iii) synergy between IL-17 and type 1 interferons in the generation and function of tertiary lymphoid structures and the associated follicular helper T-cell responses. Evaluation of these autoimmune-related pathways in TB pathogenesis provides a new perspective on recent developments in TB research

Basic science232. Certolizumab pegol prevents pro-inflammatory alterations in endothelial cell function

Background: Cardiovascular disease is a major comorbidity of rheumatoid arthritis (RA) and a leading cause of death. Chronic systemic inflammation involving tumour necrosis factor alpha (TNF) could contribute to endothelial activation and atherogenesis. A number of anti-TNF therapies are in current use for the treatment of RA, including certolizumab pegol (CZP), (Cimzia ®; UCB, Belgium). Anti-TNF therapy has been associated with reduced clinical cardiovascular disease risk and ameliorated vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to investigate the mechanisms underpinning CZP effects on TNF-activated human endothelial cells. Methods: Human aortic endothelial cells (HAoECs) were cultured in vitro and exposed to a) TNF alone, b) TNF plus CZP, or c) neither agent. Microarray analysis was used to examine the transcriptional profile of cells treated for 6 hrs and quantitative polymerase chain reaction (qPCR) analysed gene expression at 1, 3, 6 and 24 hrs. NF-κB localization and IκB degradation were investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was conducted to detect microparticle release from HAoECs. Results: Transcriptional profiling revealed that while TNF alone had strong effects on endothelial gene expression, TNF and CZP in combination produced a global gene expression pattern similar to untreated control. The two most highly up-regulated genes in response to TNF treatment were adhesion molecules E-selectin and VCAM-1 (q 0.2 compared to control; p > 0.05 compared to TNF alone). The NF-κB pathway was confirmed as a downstream target of TNF-induced HAoEC activation, via nuclear translocation of NF-κB and degradation of IκB, effects which were abolished by treatment with CZP. In addition, flow cytometry detected an increased production of endothelial microparticles in TNF-activated HAoECs, which was prevented by treatment with CZP. Conclusions: We have found at a cellular level that a clinically available TNF inhibitor, CZP reduces the expression of adhesion molecule expression, and prevents TNF-induced activation of the NF-κB pathway. Furthermore, CZP prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions and measurement of microparticles has potential as a novel prognostic marker for future cardiovascular events in this patient group. Disclosure statement: Y.A. received a research grant from UCB. I.B. received a research grant from UCB. S.H. received a research grant from UCB. All other authors have declared no conflicts of interes

Construction, Verification and Experimental Use of Two Epitope-Tagged Collections of Budding Yeast Strains

A major challenge in the post-genomic era is the development of experimental approaches to monitor the properties of proteins on a proteome-wide level. It would be particularly useful to systematically assay protein subcellular localization, post-translational modifications and protein–protein interactions, both at steady state and in response to environmental stimuli. Development of new reagents and methods will enhance our ability to do so efficiently and systematically. Here we describe the construction of two collections of budding yeast strains that facilitate proteome-wide measurements of protein properties. These collections consist of strains with an epitope tag integrated at the C-terminus of essentially every open reading frame (ORF), one with the tandem affinity purification (TAP) tag, and one with the green fluorescent protein (GFP) tag. We show that in both of these collections we have accurately tagged a high proportion of all ORFs (approximately 75% of the proteome) by confirming expression of the fusion proteins. Furthermore, we demonstrate the use of the TAP collection in performing high-throughput immunoprecipitation experiments. Building on these collections and the methods described in this paper, we hope that the yeast community will expand both the quantity and type of proteome level data available

Alforria, liberdade e cidadania: o problema da fundamentação legal da manumissão no Antigo Regime ibérico

The purpose of this article is to contribute to the understanding of the legal reasoning of manumission, the root of the systems of slavery developed in the Iberian Atlantic area. Considering the normative order of the civil law, typical of the Old Regime, aims to return some topics already covered by historiography, trying to move forward with respect to the implications of liberation in terms of obtaining citizenship and modalities effectively included by the written law.El propósito de este artículo es contribuir hacia la comprensión de los fundamentos jurídicos de la manumisión, la piedra angular de la esclavitud desarrollada en la zona atlántica ibérica. Teniendo en cuenta el orden normativo del Antiguo Régimen y recuperando algunos de los temas ya cubiertos por la historiografía, buscase avanzar analíticamente con respecto a las implicaciones de la liberación en términos de obtención de la ciudadanía y de las modalidades efectivamente establecidas por el derecho positivo. [pt] O propósito do artigo é contribuir para o entendimento da fundamentação jurídica da manumissão, pedra angular dos regimes de escravidão desenvolvidos pelos ibéricos no espaço atlântico. Considerando a ordem normativa do Antigo Regime, pautada na cultura do direito comum, procura-se retomar certos tópicos já abordados pela historiografia, tratando de avançar com respeito às implicações da libertação em termos de obtenção da cidadania e às modalidades efetivamente compreendidas pelo direito positivo