Dppa3 is a marker of pluripotency and has a human homologue that is expressed in germ cell tumours

We identified a transcript named 11M2 on the basis of its strong male-specific expression pattern in the developing mouse gonad. 11M2 was found to be expressed by gonad primordial germ cells (PGCs) of both sexes and down-regulated in female PGCs as they enter prophase I of the first meiotic division, similar to the expression of Oct4. Mouse EST analysis revealed expression only in early-stage embryos, embryonic stem cells and pre-meiotic germ cells. 11M2 corresponds to a recently reported gene variously known as PGC7, stella or Dppa3. We have identified the human orthologue of Dppa3 and find by human EST analysis that it is expressed in human testicular germ cell tumours but not in normal human somatic tissues. The expression patterns of mouse and human DPPA3, in undifferentiated embryonic cells, embryonic germ cells and adult germ cell tumours, together suggest a role for this gene in maintaining cell pluripotentiality.

Bubble load measurement in the pulp zone of industrial flotation machines - a new device for determining the froth recovery of attached particles

A new device has been developed to directly measure the bubble loading of particle-bubble aggregates in industrial flotation machines, both mechanical flotation cells as well as flotation column cells. The bubble loading of aggregates allows for in-depth analysis of the operating performance of a flotation machine in terms of both pulp/collection zone and froth zone performance. This paper presents the methodology along with an example showing the excellent reproducibility of the device and an analysis of different operating conditions of the device itself. (C) 2004 Elsevier B.V All rights reserved.

Functional memory CD8+ T cells can be generated in vivo without evident T help

Synthetic cytotoxic T cell (CTL) epitope peptides provide an effective and safe means of vaccination against cancers and viruses, as these peptides can induce specific CD8+ effector T cells in vivo. However, the effector CD8+ T cells induced by the minimal CTL epitope peptides do not last past about 3 weeks after the induction and no functional memory CD8+ T cells are generated. It is held that simultaneous induction of CD4+ T cells by incorporating peptides containing T-helper epitopes in the vaccine at the time of primary vaccination are necessary for the induction of long-lived functional memory CD8+ T cells. We now report that, surprisingly, incorporation of medium length (>20 AA) peptides devoid of detectable T-helper epitopes in a minimal CTL epitope-based vaccine can also induce long-lasting! functional rumour antigen specific memory CD8+ T cells that are capable of promoting protection against tumour challenge. This observation may have implications for the formulation of therapeutic anti-cancer and anti-virus peptide vaccines where a strong induction of CD4 T help would be undesirable. (C) 2004 Elsevier Ltd. All rights reserved.

Germ cells enter meiosis in a rostro-caudal wave during development of the mouse ovary

Germ cells in the mouse embryo remain undifferentiated until about 13.5 days post-coitum (dpc), when male germ cells enter mitotic arrest and female germ cells enter meiosis. The molecular signals and transcriptional control mechanisms governing the differential fate of germ cells in males and females remain largely unknown. In order to gain insights into the behavior of germ cells around this period and into likely mechanisms controlling entry into meiosis, we have studied by wholemount in situ hybridization the expression pattern of two germ cell-specific markers, Oct4 and Sycp3, during mouse fetal gonad development. We observed a dynamic wave of expression of both genes in developing ovaries, with Oct4 expression being extinguished in a rostro-caudal wave and Sycp3 being upregulated in a corresponding wave, during the period 13.5-15.5 dpc. These results indicate that entry into meiosis proceeds in a rostro-caudal progression, in turn suggesting that somatically derived signals may contribute to the control of germ cell entry into meiosis in developing ovaries. (C) 2004 Wiley-Liss, Inc.

Identifying the molecular phenotype of renal progenitor cells

Although many of the molecular interactions in kidney development are now well understood, the molecules involved in the specification of the metanephric mesenchyme from surrounding intermediate mesoderm and, hence, the formation of the renal progenitor population are poorly characterized. In this study, cDNA microarrays were used to identify genes enriched in the murine embryonic day 10.5 (E10.5) uninduced metanephric mesenchyme, the renal progenitor population, in comparison with more rostral derivatives of the intermediate mesoderm. Microarray data were analyzed using R statistical software to determine accurately genes differentially expressed between these populations. Microarray outliers were biologically verified, and the spatial expression pattern of these genes at E10.5 and subsequent stages of early kidney development was determined by RNA in situ hybridization. This approach identified 21 genes preferentially expressed by the E10.5 metanephric mesenchyme, including Ewing sarcoma homolog, 14-3-3 theta, retinoic acid receptor-alpha, stearoyl-CoA desaturase 2, CD24, and cadherin-11, that may be important in formation of renal progenitor cells. Cell surface proteins such as CD24 and cadherin-11 that were strongly and specifically expressed in the uninduced metanephric mesenchyme and mark the renal progenitor population may prove useful in the purification of renal progenitor cells by FACS. These findings may assist in the isolation and characterization of potential renal stem cells for use in cellular therapies for kidney disease.

Functional maturation of isolated neural progenitor cells from the adult rat hippocampus

Although neural progenitor cells (NPCs) may provide a source of new neurons to alleviate neural trauma, little is known about their electrical properties as they differentiate. We have previously shown that single NPCs from the adult rat hippocampus can be cloned in the presence of heparan sulphate chains purified from the hippocampus, and that these cells can be pushed into a proliferative phenotype with the mitogen FGF2 [Chipperfield, H., Bedi, K.S., Cool, S.M. & Nurcombe, V. (2002) Int. J. Dev. Biol., 46, 661-670]. In this study, the active and passive electrical properties of both undifferentiated and differentiated adult hippocampal NPCs, from 0 to 12 days in vitro as single-cell preparations, were investigated. Sparsely plated, undifferentiated NPCs had a resting membrane potential of approximate to -90 mV and were electrically inexcitable. In > 70%, ATP and benzoylbenzoyl-ATP evoked an inward current and membrane depolarization, whereas acetylcholine, noradrenaline, glutamate and GABA had no detectable effect. In Fura-2-loaded undifferentiated NPCs, ATP and benzoylbenzoyl-ATP evoked a transient increase in the intracellular free Ca2+ concentration, which was dependent on extracellular Ca2+ and was inhibited reversibly by pyridoxalphosphate-6-azophenyl-2'-4'-disulphonic acid (PPADS), a P2 receptor antagonist. After differentiation, NPC-derived neurons became electrically excitable, expressing voltage-dependent TTX-sensitive Na+ channels, low- and high-voltage-activated Ca2+ channels and delayed-rectifier K+ channels. Differentiated cells also possessed functional glutamate, GABA, glycine and purinergic (P2X) receptors. Appearance of voltage-dependent and ligand-gated ion channels appears to be an important early step in the differentiation of NPCs.

Modelling of entrainment in industrial flotation cells: the effect of solids suspension

Solids concentration and particle size distribution gradually change in the vertical dimension of industrial flotation cells, subject primarily to the flotation cell size and design and the cell operating conditions. As entrainment is a two-step process and involves only the suspended solids in the top pulp region near the pulp-froth interface, the solids suspension characteristics have a significant impact on the overall entrainment. In this paper, a classification function is proposed to describe the state of solids suspension in flotation cells, similar to the definition of degree of entrainment for classification in the froth phase found in the literature. A mathematical model for solids suspension is also developed, in which the classification function is expressed as an exponential function of the particle size. Experimental data collected from three different Outokumpu tank flotation cells in three different concentrators are well fitted by the proposed exponential model. Under the prevailing experimental conditions, it was found that the solids content in the top region was relatively independent of cell operating conditions such as froth height and air rate but dependent on the cell size. Moreover, the results obtained from the total solids tend to be similar to those from a particular gangue mineral and hence may be applied to all minerals in entrainment calculation. (C) 2004 Elsevier Ltd. All rights reserved.

Alterations in the phenotype of neocortical pyramidal cells in the Dyrk1A+/- mouse

The gene encoding the dual-specificity tyrosine-regulated kinase DYRK1A maps to the chromosomal segment HSA21q22.2, which lies within the Down syndrome critical region. The reduction in brain size and behavioral defects observed in mice lacking one copy of the murine homologue Dyrk1A (Dyrk1A+/-) support the idea that this kinase may be involved in monosomy 21 associated mental retardation. However, the structural basis of these behavioral defects remains unclear. In the present work, we have analyzed the microstructure of cortical circuitry in the Dyrk1A+/- mouse and control littermates by intracellular injection of Lucifer Yellow in fixed cortical tissue. We found that labeled pyramidal cells were considerably smaller, less branched and less spinous in the cortex of Dyrk1A+/- mice than in control littermates. These results suggest that Dyrk1A influences the size and complexity of pyramidal cells, and thus their capability to integrate information. (c) 2005 Elsevier Inc. All rights reserved.

Selective transport of attached particles across the pulp-froth interface

A technique for determining the recovery of attached particles across the froth phase in flotation that relies on measuring the rate at which bubble-particle aggregates enter the froth is used to investigate the selectivity of attached particles across the froth phase. Combining these measurements with those of other techniques for determining the froth recovery of attached particles provides an insight into the different sub-processes of particle rejection in the froth phase. The results of experiments conducted in a 3 m(3) Outokumpu tank cell show that the detachment of particles from aggregates in the froth phase occurs largely at the pulp-froth interface. In particular it is shown that the pulp-froth interface selectively detaches particles from aggregates according to their physical attributes. (C) 2005 Elsevier Ltd. All rights reserved.

Modelling of entrainment in industrial flotation cells: Water recovery and degree of entrainment

Entrainment in flotation can be considered as a two-step process, including the transfer of the suspended solids in the top of the pulp region just below the pulp-froth interface to the froth phase and the transfer of the entrained particles in the froth phase to the concentrate. Both steps have a strong classification characteristic. The degree of entrainment describes the classification effect of the drainage process in the froth phase. This paper briefly reviews two existing models of degree of entrainment. Experimental data were collected from an Outokumpu 3 m(3) tank cell in the Xstrata Mt. Isa Mines copper concentrator. The data are fitted to the models and the effect of cell operating conditions including air rate and froth height on the degree of entrainment is examined on a size-by-size basis. It is found that there is a strong correlation between the entrainment and the water recovery, which is close to lineal. for the fines. The degree of entrainment decreases with increase in particle size. Within the normal range of cell operating conditions, few particles coarser than 50 mu m are recovered by entrainment. In general, the degree of entrainment increases with increase in the ail rate and decreases with increase in the froth height. Air rate and froth height strongly interact with each other and affect the entrainment process mainly via changes in the froth retention time, the froth structure and froth properties. As a result, other mechanisms such as entrapment may become important in recovering the coarse entrained particles. (c) 2005 Elsevier Ltd. All rights reserved.

Stem cells in the periodontal ligament

The ability to identify and manipulate stem cells has been a significant advancement in regenerative medicine and has contributed to the development of tissue engineering-based clinical therapies. Difficulties associated with achieving predictable periodontal regeneration, means that novel techniques such as tissue engineering need to be developed in order to regenerate the extensive soft and hard tissue destruction that results from periodontitis. One of the critical requirements for a tissue engineering approach is the delivery of ex vivo expanded progenitor populations or the mobilization of endogenous progenitor cells capable of proliferating and differentiating into the required tissues. By definition, stem cells fulfill these requirements and the recent identification of stem cells within the periodontal ligament represents a significant development in the progress toward predictable periodontal regeneration. In order to explore the importance of stem cells in periodontal wound healing and regeneration, this review will examine contemporary concepts in stem cell biology, the role of periodontal ligament progenitor cells in the regenerative process, recent developments in identifying periodontal stem cells and the clinical implications of these findings.

Selective transport of attached particles across the pulp-froth interface

A technique for determining the recovery of attached particles across the froth phase in flotation that relies on measuring the rate at which bubble-particle aggregates enter the froth is used to investigate the selectivity of attached particles across the froth phase. Combining these measurements with those of other techniques for determining the froth recovery of attached particles provides an insight into the different subprocesses of particle rejection in the froth phase. The results of experiments conducted in a 3 m3 Outokumpu tank cell show that the detachment of particles from aggregates in the froth phase occurs largely at the pulp-froth interface. In particular it is shown that the pulp-froth interface selectively detaches particles from aggregates according to their physical attributes.