Zinc oxide nanoparticles as selective killers of proliferating cells

Background: It has recently been demonstrated that zinc oxide nanoparticles (ZnO NPs) induce death of cancerous cells whilst having no cytotoxic effect on normal cells. However, there are several issues which need to be resolved before translation of zinc oxide nanoparticles into medical use, including lack of suitable biocompatible dispersion protocols and a better understanding being needed of the mechanism of their selective cytotoxic action. Methods: Nanoparticle dose affecting cell viability was evaluated in a model of proliferating cells both experimentally and mathematically. The key issue of selective toxicity of ZnO NPs toward proliferating cells was addressed by experiments using a biological model of noncancerous cells, ie, mesenchymal stem cells before and after cell differentiation to the osteogenic lineage. Results: In this paper, we report a biocompatible protocol for preparation of stable aqueous solutions of monodispersed zinc oxide nanoparticles. We found that the threshold of intracellular ZnO NP concentration required to induce cell death in proliferating cells is 0.4 ± 0.02 mM. Finally, flow cytometry analysis revealed that the threshold dose of zinc oxide nanoparticles was lethal to proliferating pluripotent mesenchymal stem cells but exhibited negligible cytotoxic effects to osteogenically differentiated mesenchymal stem cells. Conclusion: Results confirm the ZnO NP selective cytotoxic action on rapidly proliferating cells, whether benign or malignant

Percepções de professores e alunos de uma região semiárida brasileira sobre educação ambiental e água

A água se constitui em um tema muito adequado para a realização de atividades de educação ambiental. No entanto, o conhecimento prévio sobre a concepção dos envolvidos nestas atividades através de pesquisas de percepção ambiental pode ajudar a dinamizar o processo de sensibilização. Com o objetivo de entender a concepção de professores e alunos da rede pública de uma cidade de região semiárida nordestina sobre o desenvolvimento de atividades de educação ambiental relacionadas ao tema água foi aplicado um questionário, como instrumento de coleta. A análise dos dados permitiu observar os professores se dizem envolvidos em atividades de educação ambiental e desenvolvem ações para minimizar os problemas da água, no entanto, não revelam concretamente como fazem isso, além de os alunos participarem pouco dessas ações

Auditory disorders and acquisition of the ability to localize sound in children born to HIV-positive mothers

The objective of the present study was to evaluate children born to HIV-infected mothers and to determine whether such children present auditory disorders or poor acquisition of the ability to localize sound. The population studied included 143 children (82 males and 61 females), ranging in age from one month to 30 months. The children were divided into three groups according to the classification system devised in 1994 by the Centers for Disease Control and Prevention: infected; seroreverted; and exposed. The children were then submitted to audiological evaluation, including behavioral audiometry, visual reinforcement audiometry and measurement of acoustic immittance. Statistical analysis showed that the incidence of auditory disorders was significantly higher in the infected group. In the seroreverted and exposed groups, there was a marked absence of auditory disorders. In the infected group as a whole, the findings were suggestive of central auditory disorders. Evolution of the ability to localize sound was found to be poorer among the children in the infected group than among those in the seroreverted and exposed groups.University of São Paulo School Infectious Diseases Division Department of PediatricsFederal University of São Paulo Department of Speech-Language and Hearing ScienceFederal University of São Paulo Department of Pediatric InfectologyUNIFESP, Department of Speech-Language and Hearing ScienceUNIFESP, Department of Pediatric InfectologySciEL

Spheroids from equine amnion mesenchymal stem cells: an in vitro study

Background: Equine amnion mesenchymal stem cells (EAMSCs) from amnion isolated after the foal birth represented an alternative source of easy collection of mesenchymal cells used in equine regenerative medicine. Methods: These cells grown as 2D culture in α-MEM medium supplemented with EGF were differentiated in adipogenic, chondrogenic and osteogenic cells. Half a million cells as pellet were left in 15ml tubes with the same differentiation media for 20 days. After the pellets were collected, embedded in paraffin for morphological study. Results: 2D culture showed EAMSCs with an embryonic phenotype (C-kit+, CD105+, Oct-4+) and a differentiation potential in adipogenic, chondrogenic and osteogenic multipotent cells. By a reproducible method of 3D culture, at day 20 the Authors evidenced a formation of small aggregated spheroids gradually gathering. In cross sections the surface of spheroid evidenced flattened cells embedded in a red matrix by Alizarin staining and occasionally a core of calcium precipitation. A network of apoptotic or necrotic cells in a not mineralized matrix was present into the center of nodules. The 3D spheroids appeared larger (mean diameter of 605±53 µm for gathering spheroids and 1486±79 µm for spheroids already gathered) than those from standard monolayer cultures (mean diameter of 200 ± 73 µm). Conclusions: EAMSCs cultured in 3D method preserve their in vitro multipotent differentiation than adherent 2D culture method. These EAMSCs included in extracellular matrix not mineralized at day 20 seem to be a good source of MSCs for tissue repair and regeneration in equine medicine

3-D Fibrin Scaffold Improves Stemness of Human Peripheral Blood Endothelial Progenitor Cells

Aims Fibrin is a natural biopolymer appealing for cell-based regenerative therapies, because it can support growth, migration and differentiation of different cell types. Endothelial progenitor cells (EPC) represent a very interesting alternative cell source for mature endothelial cells; the fact that can easily isolated from the peripheral blood, thereby eliminating donor morbidity, makes them ideal in applications in the field of regenerative medicine. We have demonstrated that fibrin can support EPC viability and growth. Aim of this study was to evaluate if fibrin can affect EPC differentiation and stem cell markers expression. Methods Fibrin was prepared mixing commercially available (Kedrion S.p.A. Lucca, Italy) fibrinogen (9 mg/ml) and thrombin (25 U/ml). Clot ultrastructure was investigated by scanning electron microscopy (SEM) and cryogenic SEM (CRYO-SEM) to measure fibre diameter and density. Clot elasticity was evaluated by atomic force microscopy (AFM), measuring the tip-sample force by cantilever displacement. EPC were obtained from peripheral blood and cultured on fibrin at the concentration of 1x106cell/cm2. Fibronectin coating was used as a control. Metabolic activity was assessed after 7 and 14 days by WST1 assay and viability by confocal microscopy (calcein incorporation). The expression of both endothelial (CD31, KDR, vWF, Ve-Cadherin) and stem cell markers (nanog, oct-4) was assessed by flow cytometry, confocal microscopy and Real Time RT-PCR. Results SEM analysis revealed a nanometric fibrous structure, with mean fiber diameter of 165?4 nm and mean density of 95.9?0.2 %. CRYO-SEM suggested a reticulate structure with mesh-size up to 10 ?m. Fibrin clot elasticity was 1.78 MPa, as in literature. WST1 assay showed that fibrin increased EPC metabolic activity as compared to fibronectin (fibrin: 0.606?0.056 a.u. vs. fibronectin: 0.311?0.067). Calcein staining demonstrated that EPC were still viable at 14 days. Flow cytometry showed the expression of endothelial markers (CD31=41.8?8.4%; vWF=32.3?3.0%; KDR=89.3?3.7%; VE-Cadherin=41.2?3.8%), confirmed also by confocal microscopy and Real Time RT-PCR. Interestingly, nanog and oct-4 (embryonic stem cell markers) expression was significantly greater on fibrin (p<0.001) as compared to fibronectin. Conclusions These findings suggest that fibrin it is not only a suitable scaffold for EPC growth and viability but also induces EPC differentiation. The observation that Nanog, known as the most important marker of stemness, is maintained longer than on fibronectin, may offer a surplus value to stem cell-based therapies

Well Design Challenges in Geothermal Energy Applications

Geothermal resources represent precious energy sources to ensure sustainable power generation. As proposed in the majority of the future sustainable energy scenarios, geothermal energy exploitation is going to play a significant role in the energy mix to meet carbon neutrality target. Upon the different technologies involved, geothermal wells constitute the core and turning point for proper fluid/heat mining. Indeed, the number of suitable candidates for geothermal applications could be significantly enhanced by overcoming a series of wells related technological issues. Therefore, the object of this work is to provide a general overview of the principal challenges that characterized well design and construction in geothermal applications which are mainly related to the type of geological system and its relative temperature level. As a matter of fact, reservoir temperature guides most of the choices referring to geothermal systems not only in the selection of the final energy application purpose (direct use, power generation, combined heat and power) but also in well design definition. Based on temperature range, geothermal fields are usually grouped in enthalpy classes (low, medium and high) referring to fields characterized by similar energy potential. From a well design and construction perspective, the low and medium enthalpy classes, in the range of temperature lower than 150 °C, do not present specific criticalities. On the contrary, high enthalpy scenarios, for temperatures higher than 170 °C, present many challenges for most of the current drilling and completion technologies. Even though some field applications exist in high/ultra-high enthalpy scenarios, they still present an elevated risk of failure. Therefore, dedicated studies shall be conducted for all the elements involved in the well construction process such as: drilling fluids, cement slurry, metallurgy, drilling and completion equipment to properly account for their specific technical limitations. In this framework, a clear picture of the actual technical gaps constitutes the starting point for current and next research activities. In the close future, the growing interest in geothermal applications will surely boost the born and development of dedicated tools to unlock the enormous potential of geothermal energy

Diffuse glioblastoma resembling acute hemorrhagic leukoencephalitis

We report the case of a young man with sudden onset of diplopia after an upper respiratory tract infection. Based on the first radiological findings acute hemorrhagic leukoencephalitis, a variant of acute disseminated encephalomyelitis, was suspected and treatment with high dose intravenous dexamethasone was started but it was stopped for intolerance. The patient clinically worsened, developing gait instability, ataxia and ophthalmoplegia; brain MRI performed 20 days later showed severe progression of the disease with subependymal dissemination. After brain biopsy of the right temporal lesion the histological diagnosis was glioblastoma. These findings suggest that MRI features of acute hemorrhagic leukoencephalitis may dissimulate the diagnosis of diffuse glioma/glioblastoma. This case underscores the importance of considering diffuse glioma in the differential diagnosis of atypical signs and symptoms of acute hemorrhagic leukoencephalitis and underlines the relevant role of integrating neuroradiologic findings with neuropathology

Pleiotropic antitumor effects of the pan-HDAC inhibitor ITF2357 against c-Myc-overexpressing human B-cell non-Hodgkin lymphomas,

Histone deacetylases (HDAC) extensively contribute to the c-Myc oncogenic program, pointing to their inhibition as an effective strategy against c-Myc-overexpressing cancers. We, thus, studied the therapeutic activity of the new-generation pan-HDAC inhibitor ITF2357 (Givinostat®) against c-Myc-overexpressing human B-cell non-Hodgkin lymphomas (B-NHLs). ITF2357 anti-proliferative and pro-apoptotic effects were analyzed in B-NHL cell lines with c-Myc translocations (Namalwa, Raji and DOHH-2), stabilizing mutations (Raji) or post-transcriptional alterations (SU-DHL-4) in relationship to c-Myc modulation. ITF2357 significantly delayed the in vitro growth of all B-NHL cell lines by inducing G1 cell-cycle arrest, eventually followed by cell death. These events correlated with the extent of c-Myc protein, but not mRNA, downregulation, indicating the involvement of post-transcriptional mechanisms. Accordingly, c-Myc-targeting microRNAs let-7a and miR-26a were induced in all treated lymphomas and the cap-dependent translation machinery components 4E-BP1, eIF4E and eIF4G, as well as their upstream regulators, Akt and PIM kinases, were inhibited in function of the cell sensitivity to ITF2357, and, in turn, c-Myc downregulation. In vivo, ITF2357 significantly hampered the growth of Namalwa and Raji xenografts in immunodeficient mice. Noteworthy, its combination with suboptimal cyclophosphamide, achieved complete remissions in most animals and equaled or even exceeded the activity of optimal cyclophosphamide. Collectively, our findings provide the rationale for testing the clinical advantages of adding ITF2357 to current therapies for the still very ominous c-Myc-overexpressing lymphomas. They equally provide the proof-of-concept for its clinical evaluation in rational combination with the promising inhibitors of B-cell receptor and PI3K/Akt/mTOR axis currently in the process of development