Melanoma contains CD133 and ABCG2 positive cells with enhanced tumorigenic potential

The failure to eradicate most cancers and in particular melanoma may be as fundamental as a misidentification of the target. The identification of cancer stem/initiating cells within the tumour population with a crucial role for tumour formation may open new pharmacological perspectives. Our data show three main novelties for human melanoma: firstly, melanoma biopsy contains a subset of cells expressing CD133 (CD133+) and the latter is able to develop a Mart-1 positive tumour in NOD-SCID mice. Secondly, the WM115, a human melanoma cell line, has been found to express both CD133 and ABCG2 markers. This cell line grows as floating spheroids, expresses typical progenitors and mature neuronal/oligodendrocyte markers and is able to transdifferentiate into astrocytes or mesenchymal lineages under specific growth conditions. As in xenografts generated with CD133+ biopsy melanoma cells, those produced by the cell line displayed lower levels of CD133 and ABCG2. Thirdly, the WM115 cells express the most important angiogenic and lymphoangiogenic factors such as notch 4, prox1 and podoplanin which can cooperate in the development of the tumourigenic capability of melanoma in vivo. Therefore, in this study, we demonstrate the presence of stem/initiating subsets in melanoma both in biopsy and in an established melanoma cell line grown in vitro and in xenografts. Interestingly, considering that melanoma gives metastasis primarily through lymphatic vessels, herein, we demonstrated that a melanoma cell line expresses typical lymphoangiogenic factors

Air pollution biomonitoring in an urban-industrial setting (Taranto, Italy) using Mediterranean plant species

This study presents the first report on the elemental composition of five Mediterranean plant species (Pinus pinaster, Eucalyptus camaldulensis, Nerium oleander, Olea europaea and Pittosporum heterophyllum) to trace industrial emissions in Taranto, Italy, a mixed-use industrial and urban setting. Potential metal sources include vehicular traffic, steel and cement plants, and a petrochemical refinery. Samples were collected from 29 sites covering the Tamburi-Lido Azzurro neighbourhood and the historical quarter Città Vecchia-Borgo. High concentrations of toxic metals were observed in all samples, with marked inter-species variability. Model based clustering identified two distinct groups, one dominated by pine needles with higher metal concentrations than the other group composed of the other four plant species. The contamination factor (CF) and pollution load index (PLI) indices which use background samples to standardise the level of pollution, were used to remove species effect allowing for direct site comparison. Spatial analysis of CF and PLI data identified pollution hotspots near industrial areas and major roads, with areas of little to no air pollution near green spaces. Statistical analysis of the CFs revealed the contribution of different sources to element emissions. Ni and Cr were primarily emitted from the steel plant and petrochemical refinery, while Fe and Al were associated with road traffic emissions, and geogenic elements Ca, Mg, K, and Na were linked to marine spray and Saharan dust. This study demonstrates that combining multiple plant species with pollution indices can be a cost-effective biomonitoring approach for assessing air pollution and creating a high-density spatial monitoring network

Unique expression and\u2028localization of aquaporin- 4 and aquaporin-9 in murine and human neural stem\u2028cells and in their glial progeny

Aquaporins (AQP) are water channel proteins that play important roles in the regulation of water homeostasis in physiological and pathological conditions. AQP4 and AQP9, the main aquaporin subtypes in the brain, are expressed in the adult forebrain subventricular zone (SVZ), where neural stem cells (NSCs) reside, but little is known about their expression and role in the NSC population, either in vivo or in vitro. Also, no reports are available on the presence of these proteins in human NSCs. We performed a detailed molecular and phenotypical characterization of different AQPs, and particularly AQP4 and AQP9, in murine and human NSC cultures at predetermined stages of differentiation. We demonstrated that AQP4 and AQP9 are expressed in adult murine SVZ-derived NSCs (ANSCs) and that their levels of expression and cellular localization are differentially regulated upon ANSC differentiation into neurons and glia. AQP4 (but not AQP9) is expressed in human NSCs and their progeny. The presence of AQP4 and AQP9 in different subsets of ANSC-derived glial cells and in different cellular compartments suggests different roles of the two proteins in these cells, indicating that ANSC-derived astrocytes might maintain in vitro the heterogeneity that characterize the astrocyte-like cell populations in the SVZ in vivo. The development of therapeutic strategies based on modulation of AQP function relies on a better knowledge of the functional role of these channels in brain cells. We provide a reliable and standardized in vitro experimental model to perform functional studies as well as toxicological and pharmacological screenings

Expression and phosphorylation of δ-CaM kinase II in cultured Alzheimer fibroblasts

Dysregulation of calcium homeostasis is among the major cellular alterations in Alzheimer’s disease (AD).We studied Ca2+/calmodulindependent protein kinase II (CaM kinase II), one of the major effectors regulating neuronal responses to changes in calcium fluxes, in cultured skin fibroblasts from subjects with sporadic AD.We found, by using PCR andWestern analysis, that human fibroblasts express the -isoform of this kinase, and that CaM kinase II is the major Ca2+/calmodulin-dependent kinase in these cells. Protein expression level of the kinase was not significantly different in AD fibroblasts. However, the total activity of the kinase (stimulated by Ca2+/calmodulin) was significantly reduced in AD cell lines, whereas Ca2+-independent activity was significantly enhanced. The percent autonomy of the kinase (Ê2+-independent/Ca2+-dependent activity) in AD cell lines was 62.8%, three-fold the corresponding percentage in control fibroblasts. The abnormal calcium-independent activity was not due to enhanced basal autophosphorylation of Thr287. The observed abnormalities, if present in brain tissue, may be implicated either in dysfunction of neuroplasticity and cognitive functions or in dysregulation of cell cycle

Efficient in vitro labeling of human neural precursor cells with superparamagnetic iron oxide particles: relevance for in vivo cell tracking

Recent studies have raised appealing possibilities of replacing damaged or lost neural cells by transplanting in vitro-expanded neural precursor cells (NPCs) and/or their progeny. Magnetic resonance (MR) tracking of superparamagnetic iron oxide (SPIO)-labeled cells is a noninvasive technique to track transplanted cells in longitudinal studies on living animals. Murine NPCs and human mesenchymal or hematopoietic stem cells can be efficiently labeled by SPIOs. However, the validation of SPIO-based protocols to label human neural precursor cells (hNPCs) has not been extensively addressed. Here, we report the development and validation of optimized protocols using two SPIOs (Sinerem and Endorem) to label human hNPCs that display bona fide stem cell features in vitro. A careful titration of both SPIOs was required to set the conditions resulting in efficient cell labeling without impairment of cell survival, proliferation, self-renewal, and multipotency. In vivo magnetic resonance imaging (MRI) combined with histology and confocal microscopy indicated that low numbers (5 x 10(3) to 1 x 10(4)) of viable SPIO-labeled hNPCs could be efficiently detected in the short term after transplantation in the adult murine brain and could be tracked for at least 1 month in longitudinal studies. By using this approach, we also clarified the impact of donor cell death to the MR signal. This study describes a simple protocol to label NPCs of human origin using SPIOs at optimized low dosages and demonstrates the feasibility of noninvasive imaging of labeled cells after transplantation in the brain; it also evidentiates potential limitations of the technique that have to be considered, particularly in the perspective of neural cell-based clinical applications

Expression and phosphorylation of δ-CaM kinase II in cultured Alzheimer fibroblasts

Dysregulation of calcium homeostasis is among the major cellular alterations in Alzheimer’s disease (AD).We studied Ca2+/calmodulindependent protein kinase II (CaM kinase II), one of the major effectors regulating neuronal responses to changes in calcium fluxes, in cultured skin fibroblasts from subjects with sporadic AD.We found, by using PCR andWestern analysis, that human fibroblasts express the -isoform of this kinase, and that CaM kinase II is the major Ca2+/calmodulin-dependent kinase in these cells. Protein expression level of the kinase was not significantly different in AD fibroblasts. However, the total activity of the kinase (stimulated by Ca2+/calmodulin) was significantly reduced in AD cell lines, whereas Ca2+-independent activity was significantly enhanced. The percent autonomy of the kinase (%Ca2+-independent/Ca2+-dependent activity) in AD cell lines was 62.8%, three-fold the corresponding percentage in control fibroblasts. The abnormal calcium-independent activity was not due to enhanced basal autophosphorylation of Thr287. The observed abnormalities, if present in brain tissue, may be implicated either in dysfunction of neuroplasticity and cognitive functions or in dysregulation of cell cycl

Expression and phosphorylation of delta-CaM kinase II in cultured Alzheimer fibroblasts.

10nonenoneCAVAZZIN C; BONVICINI C; NOCERA A; RACCHI M; KASAHARA J; TARDITO D; M. GENNARELLI; GOVONI S; RACAGNI G; POPOLI M.Cavazzin, C; Bonvicini, C; Nocera, A; Racchi, M; Kasahara, J; Tardito, D; Gennarelli, Massimo; Govoni, S; Racagni, G; Popoli, M

CD133 postive cellular population in human melanoma

The failure to eradicate most cancers and in particular melanoma may be as fundamental as a misidentification of the target (1). In this study, we demonstrate the presence of stem/initiating subsets in melanoma both in biopsy and in an established melanoma cell line grown in vitro and in xenografts. In particular, our data show two main novelties for human melanoma: firstly melanoma biopsy contains a subset of cells expressing CD133 and CD133+ subset cells able to develop a Mart-1 positive tumor in NOD-SCID mice. Secondly the WM115, a human melanoma cell line, expressed CD133 and ABCG2. This cell line grows as floating spheroids, expresses typical progenitors and mature neuronal/oligodendrocyte markers and is able to transdifferenziate into astrocytes or mesenchymal lineages under specific growth conditions. More interesting, tumor xenografts show lower levels of CD133 and ABCG2