Protective effects of melatonin against nicotine-induced oxidative damage of kidney

Several studies demonstrated that melatonin treatment prevents tissue damage in various models of oxidative stress (1). Experiments have shown that chronic nicotine administration caused oxidant damage in various organs by increasing lipid peroxidation products and decreasing the activity of endogenous antioxidants (2). The aim of this study was to investigate the effects of melatonin treatment on nicotine-induced oxidative changes in rat kidney and to explore the possible mechanisms of action. Three groups of rats were used as controls (the first without treatment, the second with melatonin alone and the third with nicotine alone). The last group of rats was orally treated with nicotine and melatonin for 28 days. Morphological changes in kidney were evaluated by histological procedures and immunohistochemical analysis using inflammation (NFkB and IL-6) and oxidative stress (SOD, CAT and iNOS) markers. Experiments performed demonstrated that nicotine administration increases inflammation and oxidative stress. Melatonin has a protective effect against nicotine kidney toxicity through an inhibition of inflammation and consequent oxidative damage. These data suggest that melatonin supplementation effectively counteracts the deleterious effect of chronic nicotine administration on kidney and attenuates oxidative damage possibly by its anti-inflammatory and antioxidant effects.This work was supported by grants from University of Brescia (EX 60%)

How should ecohazard of micropollutants in wastewater be gauged? Using bioassays to profile alternative tertiary treatments

The research on emerging pollutants in wastewater has become a worldwide issue of increasing environmental concern, especially considering the growing interest in wastewater reuse. However, the latter implies additional post-treatment after the conventional activated sludge processes, in order to produce a safer effluent. Our work aimed at determining the efficiency of reducing the toxicity associated with organic micropollutants (OMPs) in secondary wastewater effluents, using 3 different post-treatment technologies (granular activated carbon (GAC), sand biofiltration and UV irradiation): in particular, target chemical analysis of the OMPs most commonly founded in wastewater was coupled with effect-based assays (estrogenicity and mutagenicity). While chemical analysis assessed satisfactory performances for all 3 technologies in the abatement of selected OMPs, biological assays evidenced another perspective: both GAC and sand biofilters were significantly able to make the estrogenic load plummet; however, the UV system was ineffective in estrogenicity abatement, and its effluent exhibited also a slight mutagenicity, likely due to photo-transformation by-products. These results indicate that a synergistic combination of chemical analysis and biological assays can drive to a proper gauging of post-treatment technologies, taking into account not only the removal of OMPs, but also their overall toxicityThis work was conceived within a Short Term Scientific Mission (STSM) of the Water2020 Cost Action ES1202: Conceiving Wastewater Treatment in 2020 | Energetic, environmental and economic challenges. Authors from Universidade de Santiago de Compostela belong to the Galician Competitive Research Group GRC 2013-032 and to the CRETUS Strategic Partnership (AGRUP2015/02). All these programmes are co-funded by FEDER (UE)S

3D culture of isolated cells: a fast and efficient method for optimizing their histochemical and immunocytochemical analyses

The rapid development of three-dimensional (3D) culture systems and engineered cell-based tissue models gave rise to an increasing need of new techniques, allowing the microscopic observation of cell behavior/morphology in tissue-like structures, as clearly signalled by several authors during the last decennium. With samples consisting of small aggregates of isolated cells grown in suspension, it is often difficult to produce an optimal embedded preparation that can be further successfully processed for classical histochemical investigations. In this work, we describe a new, easy to use, efficient method that enables to embed an enriched "preparation" of isolated cells/small 3D cell aggregates, without any cell stress or damage. As for after tissue-embedding procedures, the cellular blocks can be further suitably processed for efficient histochemical as well as immunohistochemical analyses, rendering more informative-and attractive-studies onto 3D cell-based culture of neo-tissues. Microsc. Res. Tech., 2015. © 2015 Wiley Periodicals, Inc

A new structural method permits the evaluation of bone quality and fracture risk in a case of rapid and progressive osteoporosis of the hand (CRPS type II)

Aims Even if densitometry is still currently considered to be the gold standard in bone quality assessment, however, the medical community has long been aware that two are the factors that increase bone weakness: bone mineralization loss and micro-architectural deterioration. In effect, without prejudice to the importance of the contribution to bone resistance provided by mineralization, it is well known that the ability of bone to resist the applied loads also depends on the structural architecture of trabeculae, an aspect that the techniques currently available are not able to take into account (Liu X.S. et al., 2009). Estimating bone density alone is not, therefore, a parameter sufficient for a complete evaluation of the load bearing capability of this tissue, which, instead, requires also the consideration of the spatial arrangement of its trabecular component. Micro-numerical models have already been developed to compute the elastic properties of 3D reconstructions of trabecular structures and to perform strength predictions. Nevertheless, a widespread clinical application of 3D methods seems unlikely in the next future, given the examination costs and computational requirements. Methods The approach followed in this work focuses on the characterization of the bone structure by numerical simulation, obtained from conventional radiographic images of suitable anatomical districts. In a few seconds, the test can quantify the possible pathological alterations of bone micro-architecture by means of a structural index (SI), which is calculated from the elastic response of the reconstructed structure and the normalized sum of gray tones, indicative of the mineralization in the region under examination (Cosmi, 2008). Results Here we present the results obtained by applying the test to a particular case of rapidly progressing osteoporosis, occurred in a Complex Regional Pain Syndrome (CRPS) of the hand. CRPS is a chronic progressive disease, formerly known as \u201creflex sympathetic dystrophy\u201d and \u201ccausalgia\u201d (now classified as CRPS type I and CRPS type II, respectively), whose pathophysiology is still currently poorly understood (de Mos et al., 2008). Conclusions The results obtained show how the developed tool may be able to open important prospective applications for the study of bone pathologies (namely osteoporotic ones), in regards to the assessment of the illness course and the efficacy of potential therapies. Moreover, interesting possibilities of application exist also in the medical forensic field, where this new method can help the physician in the quantification of the biological damage. References: Cosmi F. (2008). Method to identify the mechanical properties of a material. Patent: USA No.10509512, deposited by University of Trieste. de Mos, M., Huygen, F.J., Dieleman, J.P., Koopman, J.S., Stricker, B.H. & Sturkenboom, M.C. (2008). Medical history and the onset of complex regional pain syndrome (CRPS). Pain, 139, 458-466. Liu XS, Zhang XH, Guo XE (2009). Contributions of trabecular rods of various orientations in determining the elastic properties of human vertebral trabecular bone. Bone, 45, 158-63

Innovative models to assess Multiple Myeloma biology and impact of drugs

Tumor and its embedding microenvironment form a unique, dynamic system, largely orchestrated by cellular players, including fibroblasts and endothelial cells (EC), and surrounding extracellular matrix (ECM) with its distinctive physical, biochemical, and biomechanical properties. There is a general consensus that, beyond genetic mutations and epigenetic modifications, the dialogue that occurs between tumor and its microenvironment, through soluble factors and molecular interactions, may affect tumor cells survival, growth, proliferation, response to chemical/physical factors, and lies the basis for metastatization to distant, specific organs. Accordingly, tight links between tumor and surrounding microenvironment could determine the overall sensitivity to anti-cancer drugs and therefore represent an attractive therapeutic target. Tumor microenvironment plays a critical role also in development and progression of haematological malignancies. In this regard, Multiple Myeloma (MM) represents a paradigmatic condition. The establishment of tight links between MM plasma cells and their microenvironment underlines the need for appropriate models for studying MM biology and predicting the impact of drugs. In the present paper, we briefly summarize the role of BM microenvironment and, particularly, of MM associated angiogenesis, in MM pathogenesis, progression and prognosis. We then provide an overview of the currently available MM models, including animal models and a new three-dimensional (3D), gel-based, in vitro model of human MM microenvironment. Finally, we discuss the potential of RCCS bioreactor-based, dynamic 3D model systems (cell and tissue culture) to investigate critical aspects of human MM pathobiology and possible clinical applications. Advantages and limitations of each model, relative to MM investigation and assessment of drug sensitivity, are also considered

Microfluidic Sensor for Noncontact Detection of Cell Flow in a Microchannel

A microfluidic sensor for detection of cells flowing in a microchannel is presented. The sensor consists of a PDMS (PolyDiMethylSiloxane) layer with two planar microreservoirs connected by a microchannel. The bottom sides of the microreservoirs are faced to two sensing electrodes formed on a PCB (Printed Circuit Board). A noncontact measurement is ensured by an insulator layer between the electrodes and the fluid. Particles flowing in the microchannel cause changes in the conductivity of the narrow path formed by the fluid, producing variations in the impedance between the electrodes. A tailored electronic interface based on a DDS (Direct Digital Synthesis) device is proposed to measure the impedance variations. In the experimental tests, the cell flow is detected by changes in the effective capacitance and conductance between the electrodes. These preliminary results are promising for biological measurements such as counting and sizing of cells in different matrices

SV40 Large T Antigen Expression Driven by col2a1 Regulatory Sequences Immortalizes Articular Chondrocytes but Does Not Allow Stabilization of Type II Collagen Expression

International audienceImmortalization of chondrocytes by SV40 T Ag has often been reported to trigger the loss of expression of type II collagen, one of the main differentiation markers, although some immortalized chondrocyte lines maintaining a differentiated phenotype have also been described. Here, we show using transient cotransfections in differentiated chondrocytes that, in contrast to c-src, neither SV40 T Ag, nor c-myc, decreases col2a1 transcriptional activity. Then, we report the possibility of immortalizing rabbit articular chondrocytes by expression of SV40 T Ag controlled by the col2a1 promoter and enhancer (pCol2SV). This strategy allows one to select within a population of differentiated chondrocytes those which are able to maintain functional regulation of the col2a1 gene through long-term culture. In precrisis pCol2SV-transfected chondrocytes, all-trans-retinoic acid, a downregulator of col2a1 expression, induced apoptosis, strongly suggesting the strict control of T Ag expression by col2a1 regulatory sequences. Some pCol2SVtransfected chondrocytes were definitively immortalized, after a short crisis period. However, type II collagen synthesis was restricted to a small proportion of cells, which went on to decrease with subculture, while the proportion of cells expressing T Ag was not affected. In these postcrisis cells, T Ag remained at least partially under the control of functional col2a1 regulatory elements as assessed by all-trans-retinoic acid down-regulation