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1.
Appl Radiat Isot ; 202: 111044, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37797447

ABSTRACT

Terbium-152 is one of four terbium radioisotopes that together form a potential theranostic toolbox for the personalised treatment of tumours. As 152 Tb decay by positron emission it can be utilised for diagnostics by positron emission tomography. For use in radiopharmaceuticals and for activity measurements by an activity calibrator a high radionuclide purity of the material and an accurate and precise knowledge of the half-life is required. Mass-separation and radiochemical purification provide a production route of high purity 152Tb. In the current work, two mass-separated samples from the CERN-ISOLDE facility have been assayed at the National Physical Laboratory to investigate the radionuclide purity. These samples have been used to perform four measurements of the half-life by three independent techniques: high-purity germanium gamma-ray spectrometry, ionisation chamber measurements and liquid scintillation counting. From the four measurement campaigns a half-life of 17.8784(95) h has been determined. The reported half-life shows a significant difference to the currently evaluated half-life (ζ-score = 3.77), with a relative difference of 2.2 % and an order of magnitude improvement in the precision. This work also shows that under controlled conditions the combination of mass-separation and radiochemical separation can provide high-purity 152Tb.

2.
Biomicrofluidics ; 10(1): 014107, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26858819

ABSTRACT

The broad capacity of pluripotent human embryonic stem cells (hESC) to grow and differentiate demands the development of rapid, scalable, and label-free methods to separate living cell populations for clinical and industrial applications. Here, we identify differences in cell stiffness, expressed as cell elastic modulus (CEM), for hESC versus mesenchymal progenitors, osteoblast-like derivatives, and fibroblasts using atomic force microscopy and data processing algorithms to characterize the stiffness of cell populations. Undifferentiated hESC exhibited a range of CEMs whose median was nearly three-fold lower than those of differentiated cells, information we exploited to develop a label-free separation device based on the principles of tangential flow filtration. To test the device's utility, we segregated hESC mixed with fibroblasts and hESC-mesenchymal progenitors induced to undergo osteogenic differentiation. The device permitted a throughput of 10(6)-10(7) cells per min and up to 50% removal of specific cell types per single pass. The level of enrichment and depletion of soft, pluripotent hESC in the respective channels was found to rise with increasing stiffness of the differentiating cells, suggesting CEM can serve as a major discriminator. Our results demonstrate the principle of a scalable, label-free, solution for separation of heterogeneous cell populations deriving from human pluripotent stem cells.

3.
J Cell Physiol ; 218(2): 276-84, 2009 Feb.
Article in English | MEDLINE | ID: mdl-18803233

ABSTRACT

Suppressor of cytokine signalling-2 (SOCS2) negatively regulates the signal transduction of several cytokines. Socs2(-/-) mice show increased longitudinal skeletal growth associated with deregulated GH/IGF-1 signalling. The present study examined the role of SOCS2 in endochondral ossification and trabecular and cortical bone formation, and investigated whether pro-inflammatory cytokines associated with pediatric chronic inflammatory disorders mediate their effects through SOCS2. Seven-week-old Socs2(-/-) mice were heavier (27%; P < 0.001) and longer (6%; P < 0.001) than wild-type mice. Socs2(-/-) tibiae were longer (8%; P < 0.001) and broader (18%; P < 0.001) than that of wild-type mice, and the Socs2(-/-) mice had wider growth plates (24%; P < 0.001) with wider proliferative and hypertrophic zones (10% (P < 0.05) and 14% (P < 0.001) respectively). Socs2(-/-) mice showed increased total cross-sectional bone area (16%: P < 0.001), coupled to increased total tissue area (17%; P < 0.05) compared to tibia from wild-type mice. Socs2(-/-) mice showed increased percent bone volume (101%; P < 0.001), trabecular number (82%; P < 0.001) and trabecular thickness (11%; P < 0.001), with associated decreases in trabecular separation (19%; P < 0.001). TNFalpha exposure to growth plate chondrocytes for 48 h increased SOCS2 protein expression. Growth of metatarsals from 1-day-old Socs2(-/-) and Socs2(+/+) mice, as well as expression of Aggrecan, Collagen Type II and Collagen Type X, were inhibited by TNFalpha, with no effect of genotype. Our data indicate that physiological levels of SOCS2 negatively regulate bone formation and endochondral growth. Our results further suggest that pro-inflammatory cytokines mediate their inhibitory effects on longitudinal bone growth through a mechanism that is independent of SOCS2.


Subject(s)
Bone Development , Growth Plate/metabolism , Suppressor of Cytokine Signaling Proteins/deficiency , Tibia/growth & development , Animals , Animals, Newborn , Biomarkers/metabolism , Bone Development/drug effects , Bone Resorption/metabolism , Cell Proliferation/drug effects , Cells, Cultured , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/metabolism , Cytokines/pharmacology , Female , Gene Expression Regulation , Growth Plate/cytology , Growth Plate/drug effects , Inflammation Mediators/pharmacology , Metatarsal Bones/cytology , Mice , Organ Size/drug effects , Osteogenesis/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Suppressor of Cytokine Signaling Proteins/genetics , Suppressor of Cytokine Signaling Proteins/metabolism , Tibia/drug effects , Tibia/metabolism
4.
Oncogene ; 15(15): 1781-6, 1997 Oct 09.
Article in English | MEDLINE | ID: mdl-9362444

ABSTRACT

The products (p21) of the three mammalian H-, N- and K-ras genes play important roles in intracellular signal transduction, linking membrane receptor kinases to the nuclear pathway through raf and mitogen activated protein kinase. They are involved in the regulation of proliferation and differentiation, and activating mutations of these genes are commonly associated with human cancers. Two p21 proteins are encoded by the K-ras gene (p21K-rasA and p21K-rasB) due to alternative splicing of the last exon. While the four p21ras proteins are highly homologous, their sequences diverge significantly at the C-termini, to which distinct biochemical and perhaps even functional differences may be ascribed. However, H-, N- and K-rasB appear to be ubiquitously expressed, with little evidence of tissue-specific or developmental regulation. In contrast, we now demonstrate that the expression of K-rasA is strikingly different. K-rasA is induced during differentiation of pluripotent embryonal stem cells in vitro. Its expression during early embryogenesis is limited temporally and spatially in a tissue-specific distribution which is largely maintained as an adult. This suggests a distinct biological role for p21K-rasA.


Subject(s)
Gene Expression Regulation, Developmental , Genes, ras , Alternative Splicing/genetics , Animals , Cell Differentiation/genetics , Humans , Mice , Mice, Inbred BALB C , RNA, Messenger/genetics
5.
Scand J Immunol ; 38(6): 521-8, 1993 Dec.
Article in English | MEDLINE | ID: mdl-8256109

ABSTRACT

T cells from spleens of mice infected with the erythrocytic stages of Plasmodium chabaudi chabaudi have been analysed with respect to their expression of surface molecules CD3, CD4 and CD8 and T-cell receptor (TCR) alpha beta and gamma delta. The majority of T cells from infected mice were alpha beta TCR+. However, there was an increase of approximately 8-10-fold in the proportion and total number of gamma delta T cells. Immunocytochemical analysis of sections of spleens taken from infected C57BL/6 mice during a primary infection showed that this increase took place particularly in the non-lymphoid areas. Within the alpha beta TCR+ T-cell population, both CD4+ T cells and CD8+ T cells were represented in proportions similar to those observed in normal uninfected mice. Stimulation of splenic T cells from infected mice with P. chabaudi-infected erythrocytes in vitro resulted in a blasted cell population composed predominantly of alpha beta TCR+ T cells with no preferential expansion of gamma delta TCR+ T cells. There was no evidence of superantigen-like stimulation of T cells bearing particular V beta chains of the TCR. The representation of the different V beta chains within the population was not significantly different from that seen in uninfected mice.


Subject(s)
Immunophenotyping , Malaria/immunology , Plasmodium chabaudi/immunology , Spleen/immunology , T-Lymphocytes/immunology , Animals , Antigens, CD/analysis , Antigens, Protozoan/immunology , Antigens, Surface/immunology , Cells, Cultured , Female , Lymphocyte Activation/immunology , Lymphocyte Subsets/immunology , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Receptors, Antigen, T-Cell, alpha-beta/analysis , Receptors, Antigen, T-Cell, gamma-delta/analysis
6.
Mol Microbiol ; 6(9): 1211-7, 1992 May.
Article in English | MEDLINE | ID: mdl-1588820

ABSTRACT

A new class of Bacillus thuringiensis delta-endotoxins, or insecticidal control proteins (ICPs), is defined by an apparently cryptic protein with a unique primary structure and novel entomocidal specificity for certain coleopteran and lepidopteran species. The discovery of a new group of ICPs will extend the use of this natural insecticide in integrated pest-management systems.


Subject(s)
Bacillus thuringiensis/chemistry , Bacterial Proteins/isolation & purification , Bacterial Toxins/isolation & purification , Endotoxins , Amino Acid Sequence , Animals , Bacillus thuringiensis Toxins , Bacterial Proteins/genetics , Bacterial Proteins/pharmacology , Bacterial Toxins/genetics , Bacterial Toxins/pharmacology , Base Sequence , Coleoptera , DNA, Bacterial , Hemolysin Proteins , Larva , Lepidoptera , Molecular Sequence Data , Pest Control, Biological
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