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Colorectal cancer is the fourth most common type of cancer worldwide, and adenocarcinoma cells that form the majority of colorectal tumors are markedly resistant to antineoplastic agents. Epidemiological studies have demonstrated that consumption of fruits and vegetables that are rich in polyphenols, is linked to reduced risk of colorectal cancer. In the present study, the effect of a standardized anthocyanin (ACN)‑rich extract on proliferation, apoptosis and cell cycle in the Caco-2 human colorectal cancer cell line was evaluated by trypan blue and clonogenic assays and western blot analysis of cleaved caspase‑3 and p21Waf/Cif1. The results of the current study demonstrated that the ACN extract markedly decreased Caco‑2 cell proliferation, induced apoptosis by activating caspase‑3 cleavage, and upregulated cyclin‑dependent kinase inhibitor 1 (p21Waf/Cif1) expression in a dose dependent manner. Furthermore, ACN extract was able to produce a dose‑dependent increase of intracellular reactive oxygen species (ROS) in Caco‑2 cells, together with a light increase of the cell total antioxidant status. In conclusion, the present study demonstrated that a standardized berry anthocyanin rich extract inhibited proliferation of Caco‑2 cells by promoting ROS accumulation, inducing caspase‑3 activation, and upregulating the expression of p21Waf/Cif1.

Videomicroscopy as a tool for investigating olefin gas phase co-polymerization is presented in this paper. This technique enables the simultaneous detection of the individual growth of a large number of catalyst particles. The focus is to study the kinetic behaviour of different types of Ziegler- and metallocene catalysts and to demonstrate that videomicroscopy can help to assign a given catalyst system to the appropriate model. Further, the density problem, the estimation of activation energies of single grains, the particle volume enlargement of amorphous copolymers and the comonomer effect are adressed.

Video microscopy as a tool for investigating olefin gas phase copolymerization is presented for the first time in this paper. The central theme of this work is the study of the comonomer effect shown by an unbridged metallocene catalyst supported on silica. By using video microscopy, it is possible to observe the increase in catalytic activity in terms of particle growth as well as monomer consumption. The observation that a more pronounced induction period in the particle growth profile is shown with increasing propylene concentration led us to investigate the copolymers obtained at different polymerization times using 13 C NMR analysis and single particle energy dispersive X-ray (EDX mapping). This allowed us to adapt the polymer growth and particle expansion model to the copolymerization. Besides physical causes for the comonomer effect, we wanted to determine whether the catalyst structure plays an important role in the comonomer effect. To this end we investigated two metallocenes bearing the same long bridging unit but differing in the ligand bound to the zirconium center. One metallocene bears a cyclopentadienyl ring, while the other bears an indenyl group. From a close analysis of the 13 C NMR, it is clear that both catalysts insert ethylene more easily then propylene, probably due to the long bridging unit that results in a narrower aperture angle of the ligand. In addition to this, the indenyl ligand does not allow the formation of propylene blocks even at high propylene concentration.

In this paper, a mathematical model describing olefin polymerization with metallocene catalysts is presented. It is an improvement of a previous model, the “particle growth model” (PGM) proposed by, among others, one of the authors of the present work and derives from the so-called “multigrane model” (MGM). The main differences between this work and others is a more sophisticated approach to fragmentation with respect to the MGM. Additionally, there is a more specific modeling for the unfragmented core with respect to the PGM. The numerical results obtained by the model are compared with experimental data. The results of this work allow to extend the PGM to catalysts with lower activity. The importance of those catalysts depends on the fact that high activity catalysts could bring, in some cases, too poor polymer morphology.

Free fatty acids (FFA), commonly elevated in diabetes and obesity, have been shown to impair endothelial functions and cause oxidative stress, inflammation, and insulin resistance. Anthocyanins represent one of the most important and interesting classes of flavonoids and seem to play a role in preventing cardiovascular diseases. Herein, we investigated the in vitro protective effects of cyanidin-3-O-glucoside (C3G) on cell signaling pathways in human umbilical vein endothelial cells (HUVECs) exposed to palmitic acid (PA), the most prevalent saturated FFA in circulation. Our data reported a significant augmentation of free radicals and oxidative stress in HUVECs exposed to PA for 3h, while C3G pretreatment improved intracellular redox status altered by FFA. Moreover, C3G significantly inhibited NF-κB proinflammatory pathway and adhesion molecules induced by PA, and these effects were attributed to the activation of Nrf2/EpRE pathway. In fact, C3G induced Nrf2 nuclear localization and activation of cellular antioxidant and cytoprotective genes at baseline and after PA exposure in endothelial cells. Our data confirm the hypothesis that natural Nrf2 inducers, such as C3G, might be a potential therapeutic strategy to protect vascular system against various stressors preventing several pathological conditions.

The work discussed in this paper is situated in the framework proposed by Cabré (1998a, 1998b, 1999, among others), i.e., the Communicative Theory of Terminology (CTT) which intends to explain terms “as singular units and, at the same time, similar to other communication units, within a global scheme of representation of reality.” The general purpose of this paper is to give empirical evidence of the thesis proposed by Cabré (1998, 1999, etc.), Temmerman (1997), Condamines and Rebeyrolle (1997) who discuss the univocality and monosemy of terminological units. In addition, the purpose is to show that even in texts aiming a high degree of precision, with an established and conventional form, such as legal texts (van Dijk, 1978), terminological units may show a conceptual variation. In this paper, I will specifically analyze the behavior of two terms: distinction and discrimination in a set of legal texts.

This paper describes experimental results obtained in a fixed bed reactor using a CoMoS based catalyst. The aim of this work was to produce alcohols from CO2 and H2. Conclusions from this work include: · CO2 and H2 can be used as a feed for the CoMoS catalyst to produce alcohols. · Overall, a H2/CO2 ratio of 3 (or higher) seems favorable for reaching significant conversion levels of H2 and CO2. · A relatively small temperature increase does not significantly affect the conversion, probably due to a thermodynamic equilibrium constraint. · Low temperature is better for a good alcohol / alkane ratio. Mainly methanol and ethanol are produced and relatively little propanol. Although the concept was proven, a process where CO2 and H2 are converted into alcohols/olefins is economically viable only if hydrogen can be obtained in a cost effective manner.

Neuropathic pain (NP) is a highly invalidating disease resulting as consequence of a lesion or disease affecting the somatosensory system. All the pharmacological treatments today in use give a long lasting pain relief only in a limited percentage of patients before pain reappears making NP an incurable disease. New approaches are therefore needed and research is testing stem cell usage. Several papers have been written on experimental neuropathic pain treatment using stem cells of different origin and species to treat experimental NP. The original idea was based on the capacity of stem cell to offer a totipotent cellular source for replacing injured neural cells and for delivering trophic factors to lesion site; soon the researchers agreed that the capacity of stem cells to contrast NP was not dependent upon their regenerative effect but was mostly linked to a bidirectional interaction between the stem cell and damaged microenvironment resident cells. In this paper we review the preclinical studies produced in the last years assessing the effects induced by several stem cells in different models of neuropathic pain. The overall positive results obtained on pain remission by using stem cells that are safe, of easy isolation, and which may allow an autologous transplant in patients may be encouraging for moving from bench to bedside, although there are several issues that still need to be solved.

El objetivo de este trabajo es presentar algunas reflexiones que contribuyan al conocimiento de un género académico, el artículo de investigación, a partir del análisis de algunos de los procedimientos lingüísticos que manifiestan la modalidad epistémica y evidencial. En este trabajo me propongo, en primer lugar, analizar algunos tipos de marcadores de modalidad epistémica y evidencial: verbos epistémico y evidenciales, verbos modales y semimodales, en la sección discusión/ conclusiones de artículos de investigación provenientes de dos disciplinas científicas: medicina y paleontología. En segunda instancia mi propósito es relacionar estos recursos modales con las nociones de subjetivización y gramaticalización. Intento demostrar que la modalidad es una categoría lingüística en la que existen distinciones de carácter gradual que caracterizan los marcadores lingüísticos a lo largo de una escala de menor a mayor subjetividad, relacionada con distintos grados de gramaticalización y que los recursos modales analizados se relacionan con la perspectiva temática de la disciplina.

BOLIVAR, ADRIANA Y BEKE, REBECCA (Comp.) (2011) Lectura y escritura para la investigacion. Caracas: Universidad Central de Venezuela. Consejo de Desarrollo Cientifico y Humanistico. 285 pp. ISBN 978-980-00-2685-4.

Word order may be modified with discursive purposes. In the case of marked-order sentences, the theme is embodied in grammatical devices, such as hanging topics and dislocated topics (Zubizarreta 1999, Bosque y Gutiérrez Rexach 2008). Such orderings are particularly relevant in discourse with respect to thematic prominence or information relevance. Besides, they have specific grammatical restrictions. The aim of this study is to analyze how information structures work in two specialized communication genres, and to establish relationships between information structures and textual functions in the academic genres under analysis. Our hypothesis is that information structures provide instructions for making inferences that help comprehension. The corpus consists of research papers and editorials from Medicina, an Argentinian medical journal of longstanding renown., El orden de palabras puede alterarse con fines discursivos. En el caso de las oraciones con orden marcado, el tema adopta otro tipo de articulaciones gramaticales, tales como los temas vinculantes y los temas dislocados (Zubizarreta 1999, Bosque y Gutiérrez Rexach 2008). Estas ordenaciones tienen particular relevancia en el plano discursivo, en relación con la prominencia temática o la relevancia informativa. A su vez, tienen restricciones gramaticales específicas. Este trabajo se propone analizar el funcionamiento de las estructuras informativas en dos géneros de la comunicación especializada y relacionar el tipo de estructuras informativas con las funciones textuales en los géneros académicos analizados. La hipótesis que sustenta el trabajo es que las funciones informativas constituyen instrucciones que permiten realizar inferencias que facilitan la comprensión de los enunciados. El corpus está constituido por editoriales y artículos de investigación de la revista Medicina, publicación de extensa tradición en la Argentina., A ordem das palavras pode ser alterada com finalidade discursiva. No caso de orações com ordem marcada, o tema adota um outro tipo de mecanismos gramaticais, tais como construções de tópico: os tópicos sentenciais e os tópicos discursivos (Zubizarreta 1999, Bosque e Gutiérrez Rexach 2008). Essas ordenações têm particular relevância no nível discursivo, em relação com a proeminência temática ou a relevância informativa. Ao mesmo tempo, elas têm restrições gramaticais específicas. O objetivo deste artigo é analisar o funcionamento das estruturas informativas em dois gêneros de comunicação especializada e relacionar o tipo de estruturas informativas com funções textuais nos gêneros acadêmicos analisados. A hipótese que sustenta o trabalho é que as funções de informação são instruções que permitem inferências que facilitam a compreensão dos enunciados. O corpus que serve de base ao estudo está constituído por editoriais e artigos de pesquisa do jornal Medicina, publicação com uma longa tradição na Argentina.

The archives produced by the Gonzagas, rulers of Mantua from 1328 to 1707, are one of the most complete and homogeneous collections in existence for dynastic ruling families in Europe in the Early Modern period. The Gonzaga archive was later integrated with other documents dated after the demise of the ruling family. The arrangement of the Gonzaga correspondence depended substantially on the realization, even partial, of Borsato's project. The same system has been maintained down to today, and in spite of several rearrangements and later integrations, its general features are still recognizable. In addition to the Gonzaga Archive, the notarial archives are of considerable importance. These contain an impressive quantity of records that lend themselves to research in several disciplines and are among the most important fonds of the State Archive of Mantua. Keywords:Borsato's project; Europe; Gonzaga archive; Mantua

This design report describes an up-to-date benchmark thermochemical conversion process that incorporates the latest research from NREL and other sources. Building on a design report published in 2007, NREL and its subcontractor Harris Group Inc. performed a complete review of the process design and economic model for a biomass-to-ethanol process via indirect gasification. The conceptual design presented herein considers the economics of ethanol production, assuming the achievement of internal research targets for 2012 and nth-plant costs and financing. The design features a processing capacity of 2,205 U.S. tons (2,000 metric tonnes) of dry biomass per day and an ethanol yield of 83.8 gallons per dry U.S. ton of feedstock. The ethanol selling price corresponding to this design is $2.05 per gallon in 2007 dollars, assuming a 30-year plant life and 40% equity financing with a 10% internal rate of return and the remaining 60% debt financed at 8% interest. This ethanol selling price corresponds to a gasoline equivalent price of $3.11 per gallon based on the relative volumetric energy contents of ethanol and gasoline.

This unit describes protocols for the derivation, characterization, and expansion of neural stem cell (NSC) lines from the adult mouse subvetricular zone (mNSCs), embryonic mouse brain and from the human fetal brain (hNSCs). NSCs can be isolated by enzymatic digestion of specific regions (NSCs niches) of the central nervous system (CNS) and grown in suspension. By using this methodology, NSCs form spherical clusters called neuropsheres, which are mechanically dissociated to a single-cell suspension and replated in the selective culture medium. Removal of growth factors and plating cells on an adherent substrate allows cells to differentiate into neurons, astrocytes, and oligodendrocytes, the main cell type of the CNS. Correct culturing of NSCs, according to this methodology, will allow cells to expand over 100 passages without alteration of cell karyotype, growth ability, and differentiation potential. © 2010 by John Wiley & Sons, Inc.

Background: Cell-based therapy holds great promises for demyelinating diseases. Human-derived fetal and adult oligodendrocyte progenitors (OPC) gave encouraging results in experimental models of dysmyelination but their limited proliferation in vitro and their potential immunogenicity might restrict their use in clinical applications. Virtually unlimited numbers of oligodendroglial cells could be generated from long-term self-renewing human (h)-derived neural stem cells (hNSC). However, robust oligodendrocyte production from hNSC has not been reported so far, indicating the need for improved understanding of the molecular and environmental signals controlling hNSC progression through the oligodendroglial lineage. The aim of this work was to obtain enriched and renewable cultures of hNSC-derived oligodendroglial cells by means of epigenetic manipulation. Methodology/Principal Findings: We report here the generation of large numbers of hNSC-derived oligodendroglial cells by concurrent/sequential in vitro exposure to combinations of growth factors (FGF2, PDGF-AA), neurotrophins (NT3) and hormones (T3). In particular, the combination FGF2+NT3+PDGF-AA resulted in the maintenance and enrichment of an oligodendroglial cell population displaying immature phenotype (i.e., proliferation capacity and expression of PDGFRα, Olig1 and Sox10), limited self-renewal and increased migratory activity in vitro. These cells generate large numbers of oligodendroglial progeny at the early stages of maturation, both in vitro and after transplantation in models of CNS demyelination. Conclusions/Significance: We describe a reliable method to generate large numbers of oligodendrocytes from a renewable source of somatic, non-immortalized NSC from the human foetal brain. We also provide insights on the mechanisms underlying the pro-oligodendrogenic effect of the treatments in vitro and discuss potential issues responsible for the limited myelinating capacity shown by hNSC-derived oligodendrocytes in vivo. © 2010 Neri et al.

Understanding the physiology of human neural stem cells (hNSCs) in the context of cell therapy for neurodegenerative disorders is of paramount importance, yet large-scale studies are hampered by the slow-expansion rate of these cells. To overcome this issue, we previously established immortal, non-transformed, telencephalic-diencephalic hNSCs (IhNSCs) from the fetal brain. Here, we investigated the fate of these IhNSC's immediate progeny (i.e. neural progenitors; IhNSC-Ps) upon unilateral implantation into the corpus callosum or the hippocampal fissure of adult rat brain, 3 days after global ischemic injury. One month after grafting, approximately one fifth of the IhNSC-Ps had survived and migrated through the corpus callosum, into the cortex or throughout the dentate gyrus of the hippocampus. By the fourth month, they had reached the ipsilateral subventricular zone, CA1-3 hippocampal layers and the controlateral hemisphere. Notably, these results could be accomplished using transient immunosuppression, i.e administering cyclosporine for 15 days following the ischemic event. Furthermore, a concomitant reduction of reactive microglia (Iba1+ cells) and of glial, GFAP+ cells was also observed in the ipsilateral hemisphere as compared to the controlateral one. IhNSC-Ps were not tumorigenic and, upon in vivo engraftment, underwent differentiation into GFAP+ astrocytes, and b-tubulinIII+ or MAP2+ neurons, which displayed GABAergic and GLUTAmatergic markers. Electron microscopy analysis pointed to the formation of mature synaptic contacts between host and donor-derived neurons, showing the full maturation of the IhNSC-P-derived neurons and their likely functional integration into the host tissue. Thus, IhNSC-Ps possess long-term survival and engraftment capacity upon transplantation into the globally injured ischemic brain, into which they can integrate and mature into neurons, even under mild, transient immunosuppressive conditions. Most notably, transplanted IhNSC-P can significantly dampen the inflammatory response in the lesioned host brain. This work further supports hNSCs as a reliable and safe source of cells for transplantation therapy in neurodegenerative disorders. © 2010 Rota Nodari et al.

BACKGROUND: Neural stem cells (NSCs) represent an optimal tool for studies and therapy of neurodegenerative diseases. We recently established a v-myc immortalized human NSC (IhNSC) line, which retains stem properties comparable to parental cells. Oxygen concentration is one of the most crucial environmental conditions for cell proliferation and differentiation both in vitro and in vivo. In the central nervous system, physiological concentrations of oxygen range from 0.55 to 8% oxygen. In particular, in the in the subventricular zone niche area, it's estimated to be 2.5 to 3%. METHODOLOGY/PRINCIPAL FINDINGS: We investigated in vitro the effects of 1, 2.5, 5, and 20% oxygen concentrations on IhNSCs both during proliferation and differentiation. The highest proliferation rate, evaluated through neurosphere formation assay, was obtained at 2.5 and 5% oxygen, while 1% oxygen was most noxious for cell survival. The differentiation assays showed that the percentages of beta-tubIII+ or MAP2+ neuronal cells and of GalC+ oligodendrocytes were significantly higher at 2.5% compared with 1, 5, or 20% oxygen at 17 days in vitro. Mild hypoxia (2.5 to 5% oxygen) promoted differentiation into neuro-oligodendroglial progenitors as revealed by the higher percentage of MAP2+/Ki67+ and GalC+/Ki67+ residual proliferating progenitors, and enhanced the yield of GABAergic and slightly of glutamatergic neurons compared to 1% and 20% oxygen where a significant percentage of GFAP+/nestin+ cells were still present at 17 days of differentiation. CONCLUSIONS/SIGNIFICANCE: These findings raise the possibility that reduced oxygen levels occurring in neuronal disorders like cerebral ischemia transiently lead to NSC remaining in a state of quiescence. Conversely, mild hypoxia favors NSC proliferation and neuronal and oligodendroglial differentiation, thus providing an important advance and a useful tool for NSC-mediated therapy of ischemic stroke and neurodegenerative diseases like Parkinson's disease, multiple sclerosis, and Alzheimer's disease.

In this report we emphasize the opportunity of considering the uncommon causes of chronic GH-excess in the initial diagnostic process, such as GHRH hypersecretion, especially in the presence of ambiguous pituitary neuroimaging. This topic may have an important clinical significance in order to plan the most cost-effective diagnostic procedures and management and to avoid unnecessary pituitary neurosurgery.

ObjectiveRadiotherapy (RT) for pituitary adenomas, including GH-secreting ones, frequently leads to GH deficiency (GHD). Data on the effects of surgery alone (S) on dynamic GH secretion are limited. The aim of the study was to investigate the occurrence of GHD in acromegalic patients treated with different therapeutic options.Design and methodsFifty-six patients in remission from acromegaly, (33 F & 23 M, age: 54±13 years, body mass index (BMI): 28.4±4.1 kg/m2, 21 with adequately substituted pituitary deficiencies) treated by S alone (n=33, group 1) or followed by RT (n=23, group 2), were investigated for GHD by GHRH plus arginine testing, using BMI-adjusted cut-offs. Several metabolic and cardiovascular parameters (waist circumference, body fat percentage, blood pressure, fasting and post-oral glucose tolerance test glucose, HbA1c, insulin resistance and lipid profile) were evaluated in all the patients and 28 control subjects with known diagnosis of GHD.ResultsSerum GH peak after challenge was 8.0±9.7 μg/l, without any correlation with post-glucose GH nadir and IGF-1 levels. The GH response indicated severe GHD in 34 patients (61%) and partial GHD in 15 patients (27%). IGF-1 were below the normal range in 14 patients (25%). The frequency of GHD was similar in the two treatment groups (54% in group 1 and 70% in group 2). No significant differences in metabolic parameters were observed between acromegalic patients and controls with GHD.ConclusionsSevere GHD may occur in about 60% of patients treated for acromegaly, even when cured after S alone. Thus, a stimulation test (i.e. GHRH plus arginine) is recommended in all cured acromegalic patients, independently from previous treatment.

A 3.4-year-old girl was admitted to the Pediatric Department because of tall stature (116.0 cm, +5.1 SDS) and increased height velocity (16.3 cm/year, +6.1 SDS). Basal hormonal evaluation revealed elevated insulin-like growth factor I (IGF-I) levels (938 ng/ml, nv 40–190), prolactin (PRL) (98.0 ng/ml, nv 1.7–24.0) and mean growth hormone (GH) nocturnal concentration (147 ng/ml). Basal adrenal, gonadal and thyroid functions were normal. Hand-wrist bone age was 3.6 years. Magnetic resonance imaging revealed a macroadenoma with moderate suprasellar invasion. The adenoma was surgically removed and histological characterization confirmed the diagnosis of GH/PRL-secreting adenoma. The patient was admitted to our Endocrine Unit when 7.9 years old, because of the persistence of elevated GH, IGF-I and PRL levels, although there was a slight height velocity reduction and absence of tumor recurrence. Treatment with cabergoline was initiated, but only PRL levels normalized. Afterwards, octreotide long-acting release (LAR) was added without reaching the normalization of GH and IGF-I levels. Thus, treatment with octreotide LAR was discontinued and pegvisomant was added to cabergoline, leading to the normalization of IGF-I levels and height velocity without side effects. Other anterior pituitary functions were always normal. To conclude, treatment of pituitary gigantism with pegvisomant was effective and well tolerated in a young giant unresponsive to combined cabergoline and octreotide treatment.

Chronic fatigue syndrome (CFS) is a specific clinical condition that characterises unexplained disabling fatigue and a combination of non-specific accompanying symptoms for at least 6 months, in the absence of a medical diagnosis that would otherwise explain the clinical presentation. Other common symptoms include headaches, myalgia, arthralgia, and post-exertional malaise; cognitive difficulties, with impaired memory and concentration; unrefreshing sleep; and mood changes. Similar disorders have been described for at least two centuries and have been differently named neurasthenia, post-viral fatigue, myalgic encephalomyelitis and chronic mononucleosis. Recent longitudinal studies suggest that some people affected by chronic fatigue syndrome improve with time but that most remain functionally impaired for several years. The estimated worldwide prevalence of CFS is 0.4-1% and it affects over 800,000 people in the United States and approximately 240,000 patients in the UK. No physical examination signs are specific to CFS and no diagnostic tests identify this syndrome. The pathophysiological mechanism of CFS is unclear. The main hypotheses include altered central nervous system functioning resulting from an abnormal immune response against a common antigen; a neuroendocrine disturbance; cognitive impairment caused by response to infection or other stimuli in sentient people. The current concept is that CFS pathogenesis is a multifactorial condition. Various studies have sought evidence for a disturbance in immunity in people with CFS. An alteration in cytokine profile, a decreased function of natural killer (NK) cells, a presence of autoantibodies and a reduced responses of T cells to mitogens and other specific antigens have been reported. The observed high level of pro-inflammatory cytokines may explain some of the manifestations such as fatigue and flu-like symptoms and influence NK activity. Abnormal activation of the T lymphocyte subsets and a decrease in antibody-dependent cell-mediated cytotoxicity have been described. An increased number of CD8+ cytotoxic T lymphocytes and CD38 and HLA-DR activation markers have been reported, and a decrease in CD11b expression associated with an increased expression of CD28+ T subsets has been observed. This review discusses the immunological aspects of CFS and offers an immunological hypothesis for the disease processes.

Human neural stem cells (hNSC) represent an essential source of renewable brain cells for both experimental studies and cell replacement therapies. Their relatively slow rate of proliferation and physiological senescence in culture make their use cumbersome under some experimental and pre-clinical settings. The immortalization of hNSC with the v-myc gene (v-IhNSC) has been shown to generate stem cells endowed with enhanced proliferative capacity, which greatly facilitates the study of hNSCs, both in vitro and in vivo. Despite the excellent safety properties displayed by v-IhNSCs – which do not transform in vitro and are not tumorigenic in vivo – the v-myc gene contains several mutations and recombination elements, whose role(s) and effects remains to be elucidated, yielding unresolved safety concerns. To address this issue, we used a c-myc T58A retroviral vector to establish an immortal cell line (T-IhNSC) from the same hNSCs used to generate the original v-IhNSCs and compared their characteristics with the latter, with hNSC and with hNSC immortalized using c-myc wt (c-IhNSC). T-IhNSCs displayed an enhanced self-renewal ability, with their proliferative capacity and clonogenic potential being remarkably comparable to those of v-IhNSC and higher than wild type hNSCs and c-IhNSCs. Upon growth factors removal, T-IhNSC promptly gave rise to well-differentiated neurons, astrocytes and most importantly, to a heretofore undocumented high percentage of human oligodendrocytes (up to 23%). Persistent growth-factor dependence, steady functional properties, lack of ability to generate colonies in soft-agar colony-forming assay and to establish tumors upon orthotopic transplantation, point to the fact that immortalization by c-myc T58A does not bring about tumorigenicity in hNSCs. Hence, this work describes a novel and continuous cell line of immortalized human multipotent neural stem cells, in which the immortalizing agent is represented by a single gene which, in turn, carries a single and well characterized mutation. From a different perspective, these data report on a safe approach to increase human neural stem cells propagation in culture, without altering their basic properties. These T-IhNSC line provides a versatile model for the elucidation of the mechanisms involved in human neural stem cells expansion and for development of high throughput assays for both basic and translational research on human neural cell development. The improved proclivity of T-IhNSC to generate human oligodendrocytes propose T-IhNSC as a feasible candidate for the design of experimental and, perhaps, therapeutic approaches in demyelinating diseases.

The treatment of children affected by severe congenital neutropenia (SCN) with G-CSF strongly reduces the risk of sepsis by reversing neutropenia. However, SCN patients who respond to the treatment with the growth factor still have an elevated risk of succumbing to sepsis. Because the disease is usually caused by heterozygous mutations of ELA2, a gene encoding for neutrophil elastase (NE), we have investigated in G-CSF–responder and nonresponder patients affected by SCN the expression of polypeptides that constitute the antimicrobial machinery of these cells. In peripheral blood–derived neutrophils of patients with heterozygous mutations of ELA2 who were treated with G-CSF, NE was nearly absent as detected by immunofluorescence and immunoblotting, suggesting that production of the mutant protein interferes with normal gene expression. This defect was associated with abnormal expression of other granule-associated proteins such as myeloperoxidase, lactoferrin, cathepsin G, and human-neutrophil-peptide. Moreover, in one patient with partial response to G-CSF, we observed an impairment of neutrophil antimicrobial activity against Candida albicans, and, to a lower extent against Escherichia coli. Thereby, we propose that the treatment with G-CSF is not sufficient to correct all of the functional deficiency of neutrophils, and this might account for the consistent risk of infections observed in SCN patients.

The correct identity and functional capacity of transplanted dopamine (DA) neurons derived in vitro from embryonic stem (ES) cells is a critical factor for the development of an ES cell-based replacement therapy for Parkinson's disease. We transplanted primate Cyno-1 ES cells differentiated in vitro for 4 (progenitor ES cells) or 6 (differentiated ES cells) weeks, or control fetal primate cells into the striatum of hemi-parkinsonian rats. Partial behavioral recovery in amphetamine-induced rotation was correlated with the number of ES-derived tyrosine hydroxylase-positive (TH +) neurons in the grafts (r = 0.5, P < 0.05). Post mortem analysis of ES-derived grafts revealed TH + neurons with mature morphology, similar to fetal DA neurons, and expression of midbrain transcription factors, such as Engrailed (En) and Nurr-1. While the total number of TH + neurons was not different between the two groups, TH/En co-expression was significantly higher (> 90%) in grafts from differentiated ES cells than in grafts derived from progenitor cells (< 50%), reflecting a more heterogeneous cellular composition. Within the grafts there was an overlap between ES-derived TH + axonal arbors and clusters of primate ES-derived striatal neurons expressing brain factor 1 (Bf-1, Foxg1) and DA and cAMP-regulated phosphoprotein (DARPP-32). Such overlap was never observed for other regional transcription factors that define neighboring forebrain domains in the developing brain, such as Nkx2.1 (medial ganglionic eminence), Nkx2.2 (pallidal and diencephalic progenitors) or Pax6 (dorsal telencephalic progenitors). Despite the heterogeneity of ES-derived graft cell composition, these results demonstrate normal phenotypic specification, conserved natural axonal target selectivity and functionality of DA neurons derived from primate ES cells. © The Authors (2006).

Adaptor protein-3 (AP-3) is an ubiquitous cytoplasmic complex that shuttles cargo proteins from the trans-Golgi and a tubular-endosomal compartment to endosome-lysosome-related organelles. Lack of the beta3A subunit of this complex causes Hermansky-Pudlak syndrome type 2, an autosomal recessive disease characterized by partial albinism, prolonged bleeding tendency, and immunodeficiency. To investigate the pathogenesis of immunodeficiency, we studied natural killer (NK) cells and neutrophil functions in 2 previously unreported siblings affected by Hermansky-Pudlak type 2 syndrome. In both patients we observed a dramatic reduction of cytolytic activity of freshly isolated and of IL-2-activated NK cells. Levels of perforin were reduced in unstimulated NK cells, thereby accounting for the impairment of NK cytolitic activity. In addition, analysis of neutrophils in these patients demonstrated that intracellular elastase content was largely reduced while CD63 expression on plasma membrane was substantially increased. Taken together, these observations suggest that type 2 Hermansky-Pudlak syndrome is characterized by defects of innate immunity.

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.

It is demonstrated that modern optical methods allow for a very fast study of the behaviour of supported metallocene catalysts. By videomicroscopy several polymeric supports functionalised with cyclopentadiene units or poly(ethylene oxide) and poly(propylene oxide) chains and loaded with a metallocene were simultaneously studied. In this example it is shown that this method allows for a direct comparison of different catalysts under identical conditions in a single experiment. In addition by this experiment more detailed information on the behaviour of the supports can be obtained by studying the growth kinetics of single grains. As a second tool, laser scanning confocal fluorescence microscopy (LSCFM) was applied. Using dye-labelled supports LSCFM can be used as a very rapid tool to study the fragmentation of supports. This is demonstrated by 3D images showing the distribution of different support fragments in the polyethylene product particles. Introduction For industrial applications of metallocenes, immobilisation of the catalyst is necessary to allow for perfect product control, especially of particle size, density and form of the polyolefin particles. Therefore, many different supports for immobilisation have been investigated. Mainly they are based on inorganic materials but also organic supports have received increasing attention [1-16]. It had been shown that supports have to fulfil several requirements as to control the polymerisation process and to obtain products with a desirable morphology. The carrier has quantitatively to immobilise the metallocene to avoid leaching and reactor fouling during the polymerisation process. Typically the supports have to be particles with an average size of 50 100 μm (final particles); however, they should be able to fragment during the polymerisation into nm-sized particles (primary particles). So far the total course of olefin polymerisation and fragmentation of the supported catalyst as a part of this process have been studied by a) Buchi reactor kinetic [17-21]: measuring the monomer consumption gives information about the course of the instationary polymerisation rate with time, reaction orders and activation energy. 1 Unauthenticated Download Date | 6/27/17 8:01 PM b) Bulk polymerisation kinetics in liquid monomer: measuring the polymer formation by means of a heat flow calorimeter leads again to the instationary polymerisation rate, reaction orders and activation energy [22-24]. c) Electron microscopic kinetics: electron microtome sections (scanning electron microscopy and transmission electron microscopy) of the polymerising particles give information about polymer growth, particle fragmentation and particle expansion [22,17]. The results from these investigations led to mathematical models of the total polymerisation process as, e.g., the polymeric flow model, the multi-grain model, and the polymer growth and particle expanding model [25,26]. In this paper we introduce videomicroscopy and laser scanning confocal optical microscopy (LSCFM) as two new optical methods for determining the particle expansion kinetics and particle fragmentation. Videomicroscopy allows the simultaneous observation of many polymerizing particles in situ [27,28]. LSCFM originally used in biology and life sciences to study cells for directly investigating the distribution of a fluorescence-tagged biologically active structure [29] allows studying the fragmentation process. By incorporation of a fluorescent dye into the supports the distribution of the fragments of the support can be measured in the growing and expanding particles within a few minutes. Results and discussion Synthesis of the organic supports To show the applicability of videomicroscopy as a tool to compare different catalysts at the same time, three organic supports as carriers for a metallocene catalyst (Me2Si(2MeBenzInd)2ZrCl2) were prepared. Polystyrene (PS) [14] or nano-sized latex particles [13] with poly(ethylene oxide) (PEO) chains on their surfaces were used as carriers for the metallocene catalyst. Due to the nucleophilic character of the ether groups, the methylaluminoxane (MAO) activated metallocene complexes are immobilised by non-covalent interactions [12,13]. Especially, nano-sized (100 150 nm) latex particles have been proposed as suitable supports for metallocenes as they fragment completely within the final product during the polymerisation reaction, as shown by transmission electron microscopy and fluorescence microscopy [13,30]. The support for catalyst K-1 (Scheme 1) consists of a linear PS chain functionalised with PEO for immobilizing the active metallocene catalyst and with cyclopentadiene (CP) for allowing reversible network formation by a Diels-Alder reaction [31]. This support was proven to be a suitable carrier for metallocene catalysts in olefin polymerisation as it shows good activity in the ethylene polymerisation and PE particles with high bulk densities were obtained. The support for catalyst K-2 (Scheme 2) was also functionalised with PEO chains and CP units; however, this support consists of latex particles prepared by emulsion polymerisation [14]. In the cases of K-2 the support is aggregated not only by DielsAlder reactions between different cyclopentadienyl units but also non-covalently due to the interaction of the polar surfaces of the particles with MAO. As a third example, latex particles (Scheme 3) surface-functionalised only with PEO were applied as supports to obtain catalyst K-3. In this approach, PS-PEO block copolymers were used as surfactants in the emulsion polymerisation process [13]. 2 Unauthenticated Download Date | 6/27/17 8:01 PM The latex particles (primary particles) are only physically aggregated by polar interactions of the ether groups of the surfactant with MAO.

We report the first post-mortem analysis of two patients with Parkinson's disease who received fetal midbrain transplants as a cell suspension in the striatum, and in one case also in the substantia nigra. These patients had a favourable clinical evolution and positive 18F-fluorodopa PET scans and did not develop motor complications. The surviving transplanted dopamine neurons were positively identified with phenotypic markers of normal control human substantia nigra (n = 3), such as tyrosine hydroxylase, G-protein-coupled inward rectifying current potassium channel type 2 (Girk2) and calbindin. The grafts restored the cell type that provides specific dopaminergic innervation to the most affected striatal regions in the parkinsonian brain. Such transplants were able to densely reinnervate the host putamen with new dopamine fibres. The patients received only 6 months of standard immune suppression, yet by post-mortem analysis 3-4 years after surgery the transplants appeared only mildly immunogenic to the host brain, by analysis of microglial CD45 and CD68 markers. This study demonstrates that, using these methods, dopamine neuronal replacement cell therapy can be beneficial for patients with advanced disease, and that changing technical approaches could have a favourable impact on efficacy and adverse events following neural transplantation. © The Author (2005). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved

Paraneoplastic syndromes are uncommon diseases with different pathogenesis and clinical manifestations, correlated with neoplasms but not due to the tumor, metastasis or other distant effects. The aim of the present article is to describe the main paraneoplastic syndromes (neurological, endocrine-metabolic, rheumatological, osteo-articular, dermatological, hematological, vascular and nephrological), the associated pathogenetic theories (theory of the common embryonal sketches, theory of reactivation of the information and autoimmune theory) and the most important therapeutic approaches, on the basis of the literature. Experimental works, reviews and clinical observations, in some cases still in progress, regarding the described syndromes, their pathogenesis and their therapeutic approach have been examined. No meta-analyses regarding paraneoplastic syndromes have been published in the literature. The better described pathogenesis is the autoimmune one, characteristic of neurological, nephrologic and some dermatologic syndromes, for which the clinical and laboratory findings have been well supported. The pathogenetic theories associated with the other syndromes have been correlated on the basis of the literature. Paraneoplastic syndromes are important because their identification permits an early diagnosis of tumors and rapid treatment, with a largely improved prognosis and life expectancy for the patient. They often represent the only signal of a silent neoplasm; sometimes they precede the tumor itself. More studies are necessary for a better definition of their clinical aspects and pathogenesis and to delineate standard guidelines for a diagnostic-therapeutic approach to these diseases.

Dopamine (DA) neurons can be derived from human and primate embryonic stem (ES) cells in vitro. An ES cell-based replacement therapy for patients with Parkinson's disease requires that in vitro-generated neurons maintain their phenotype in vivo. Other critical issues relate to their proliferative capacity and risk of tumor formation, and the capability of migration and integration in the adult mammalian brain. Neural induction was achieved by coculture of primate parthenogenetic ES cells (Cyno-1) with stromal cells, followed by sequential exposure to midbrain patterning and differentiation factors to favor DA phenotypic specification. Differentiated ES cells were treated with mitomycin C and transplanted into adult immunosuppressed rodents and into a primate (allograft) without immunosuppression. A small percentageof DA neurons survived in both rodent and primate hosts for the entire term of the study (4 and 7 months, respectively). Other neuronal and glial populations derived from Cyno-1 ES cells showed, in vivo, phenotypic characteristics and growth and migration patterns similar to fetal primate transplants, and a majority of cells (>80%) expressed the forebrain transcription factor brain factor 1. No teratoma formation was observed. In this study, we demonstrate long-term survival of DA neurons obtained in vitro from primate ES cells. Optimization of differentiation, cell selection, and cell transfer is required for functional studies of ES-derived DA neurons for future therapeutic applications. ©AlphaMed Press.

Stem cells isolated from the central nervous system of both embryonic and adult mice can generate neurons and glia through the activation of different patterns of differentiation in dependence of exposure to appropriate epigenetic signals. On the other hand, environmental conditions might affect the proliferation, migration, and differentiation of these cells. We report here, for the first time, that inorganic mercury affects the proliferative and differentiative capacity of adult neuronal stem cells (ANSCs). Actually, inorganic mercury increases apoptosis in ASNC. Furthermore, in stem cell-derived astrocytes, high levels of the 70 kDa heat shock protein (HSP-70) occur, while the levels of GTP-beta-tubulin activity dramatically decrease. Interestingly, when induced to differentiate, inorganic mercury modifies morphological proprieties of astrocytes, while the neuron population is reduced. These results demonstrate that inorganic mercury produces toxicity in the ANSC-derived neuronal population and affects the biological properties of the glial-derived population.

The paper discusses the issue of feed ingredients in aquaculture as a telling example of implementation of a sustainable food safety strategy, aimed at protecting the health of next generation, under the One Health paradigm. Finfish and fishery products are a main nutrition security component as a valuable source of animal protein, particularly in developing countries. In addition, they are a critical source of essential oligo-nutrients, such as polyunsaturated fatty acids (PUFAs) and iodine. Production and consumption of fish has greatly increased in the last decade, mostly due to the growth of aquaculture. While the demand for aquaculture products continues to increase, there is the need to address consumers' concerns related to the nutritional quality and safety. In fact, both wild and farmed finfish can represent a significant source of exposure to contaminants for the consumer: noticeably, caught and farmed fish have a comparable content of nutrients and contaminants. Aquaculture feeds made of fish meal and fish oil are the main vehicle for transfer of environmental pollutants to farmed fish. The main fish contaminants (e.g., methylmercury, PCBs, PBDE) can bioaccumulate and affect development in humans. Feed ingredients as well fish species have a different liability to contamination depending, e.g., on the lipophilicity of the specific chemicals. Up-to-date risk-benefit assessments show that high intake of fish may lead to an undesirable intake of pollutants which is not sufficiently balanced by the concurrent intake of protective nutrients, such as PUFA. The use of vegetable-based feed ingredients in aquaculture has been explored from the standpoints of economic sustainability and fish productivity to a greater extent than from those of food safety and nutritional value. Available data show that vegetable oils can significantly modulate the lipid profile in fish flesh, depending on the oil and fish species. The use of vegetable ingredients can drastically reduce the accumulation of the main contaminants in fish; likewise the presence of other “unconventional” contaminants (e.g. PAHs) and the nutritional value of fish flesh could deserve more attention in the assessment of novel aquaculture feeds.

CoMoSx/K catalysts used for the synthesis of higher alcohols from syngas are usually prepared using costly ammonium tetrathiomolybdate. In an attempt to identify inexpensive and more readily available molybdenum precursors with improved performance, catalysts were prepared starting from crystalline MoS2 activated by ball-milling. This communication reports the catalytic performance of these catalysts in the hydrogenation of CO for the synthesis of higher alcohols along with catalyst characterization results.

Bevacizumab has been introduced in the management of high-grade gliomas after preliminary studies that showed an acceptable safety and a marked increase in clinico-radiological responses in comparison with second-line chemotherapy. The objective is to synthetically review the present use of bevacizumab--alone or in combination--in the context of recurrent high-grade glioma and highlight the future developments. The methodology of this study is to analyse and discuss relevant literature studies using bevacizumab in recurrent high-grade glioma. Bevacizumab may be used as single-agent therapy in recurrent high-grade glioma, with good clinico-radiological responses having little effect on survival. The open questions and developments include new MRI criteria for evaluation of response to anti-angiogenic agents, the identification of putative factors predicting response/failure of bevacizumab and the introduction of bevacizumab in first-line management of high-grade glioma.