Cellular Modifications Research Articles

Sequential air–liquid exposure of human respiratory cells to chemical and biological pollutants

Although indoor air has wide ranging effects on human health, the effects of environmental, chemical, and biological pollutants on the respiratory system are not fully understood. In order to clarify the health effects of airborne pollutant exposure, it would appear that toxicological evidence is needed to complement epidemiological observations to support by providing biological plausibility. The aim of this study is to manage air–liquid successive exposures to different pollutants such as a chemical pollutant (formaldehyde – FA), and a biological contaminant ( Aspergillus fumigatus – Asp) using our in vitro model. Human alveolar cells (A549) were exposed at the air–liquid interface in an exposure module, firstly to an environmental level of FA (50 μg/m 3) (or air) for 30 min, and 14 h later to Asp (7 × 10 8 spores/m 3) (or air) for 30 min. After 10 h post-incubation, cellular viability was assessed. Inflammation biomarkers (IL-8, MCP-1) were assayed by ELISA and by RT-PCR. Whatever the conditions, no cytotoxic effect was observed. FA followed by air exposure did not induce modification of production and expression of cytokines, confirming results with a unique FA exposure. Air followed by Asp exposure tended to induce IL-8 expression whereas IL-8 production tended to increase after FA and Asp exposure compared to FA and air exposure. The reaction of cells to sequential exposure to FA and Asp was moderate. These results show the feasibility of our model for sequential exposures to different types of environmental pollutants, allowing using it for preliminary assessment of cellular activity modification induced by airborne contaminants.

Static retention of the lumenal monotopic membrane protein torsinA in the endoplasmic reticulum

TorsinA is a membrane-associated enzyme in the endoplasmic reticulum (ER) lumen that is mutated in DYT1 dystonia. How it remains in the ER has been unclear. We report that a hydrophobic N-terminal domain (NTD) directs static retention of torsinA within the ER by excluding it from ER exit sites, as has been previously reported for short transmembrane domains (TMDs). We show that despite the NTD's physicochemical similarity to TMDs, it does not traverse the membrane, defining torsinA as a lumenal monotopic membrane protein and requiring a new paradigm to explain retention. ER retention and membrane association are perturbed by a subset of nonconservative mutations to the NTD, suggesting that a helical structure with defined orientation in the membrane is required. TorsinA preferentially enriches in ER sheets, as might be expected for a lumenal monotopic membrane protein. We propose that the principle of membrane-based protein sorting extends to monotopic membrane proteins, and identify other proteins including the monotopic lumenal enzyme cyclooxygenase 1 (prostaglandin H synthase 1) that share this mechanism of retention with torsinA.

Cyst and encystment in protozoan parasites: optimal targets for new life-cycle interrupting strategies?

Certain protozoan parasites use survival strategies to reside outside the host such as the formation of cysts. This dormant and resistant stage results from the complex process of encystment that involves diverse molecular and cellular modifications. The stimuli and changes associated with cyst biogenesis are a matter of ongoing studies in human and animal protozoan parasites such as amoeba and Giardia species because blocking every step in the encystment pathway should, in theory, interrupt their life cycles. The present review thoroughly examines this essential process in those protozoan parasites and discusses the possibility of using that information to develop new kinds of anti-parasite specific and life cycle-interrupting drugs, aimed at holding back the dissemination of these infections.

Toxicity of nano- and micro-sized silver particles in human hepatocyte cell line L02

Silver nanoparticles (Ag NPs) previously classified as antimicrobial agents have been widely used in consumers and industrial products, especially food storage material. Ag NPs used as antimicrobial agents may be found in liver. Thus, examination of the ability of Ag NPs to penetrate the liver is warranted. The aim of the study was to determine the optimal viability assay for using with Ag NPs in order to assess their toxicity to liver cells. For toxicity evaluations, cellular morphology, mitochondrial function (3-(4, 5-dimethylazol-2-yl)-2, 5-diphenyl-tetrazolium bromide, MTT assay), membrane leakage of lactate dehydrogenase (lactate dehydrogenase, LDH release assay), Oxidative stress markers (malonaldehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD)), DNA damage (single cell gel eletrophoresis, SCGE assay), and protein damage were assessed under control and exposed conditions (24 h of exposure). The results showed that mitochondrial function decreased significantly in cells exposed to Ag NPs at 25 μg·mL−1. LDH leakage significantly increased in cells exposed to Ag NPs (≥ 25 μg mL−1) while micro-sized silver particles tested displayed LDH leakage only at higher doses (100 μg·mL−1). The microscopic studies demonstrated that nanoparticle-exposed cells at higher doses became abnormal in size, displaying cellular shrinkage, and an acquisition of an irregular shape. Due to toxicity of silver, further study conducted with reference to its oxidative stress. The results exhibited significant depletion of GSH level, increase in SOD levels and lead to lipid peroxidation, which suggested that cytotoxicity of Ag NPs in liver cells might be mediated through oxidative stress.The results demonstrates that Ag NPs lead to cellular morphological modifications, LDH leakage, mitochondrial dysfunction, and cause increased generation of ROS, depletion of GSH, lipid peroxidation, oxidative DNA damage and protein damage. Though the exact mechanism behind Ag NPs toxicity is suggested oxidative stress and lipid peroxidation playing an important role in Ag NPs elicited cell membrane disruption, DNA damage, protein damage and subsequent cell death. Our preliminary data suggest that oxidative stress might contribute to Ag NPs cytotoxicity. To reveal whether apoptosis involved in Ag NPs toxicity, further studies are underway.

Characterization of the age-dependent intervertebral disc changes in rabbit by correlation between MRI, histology and gene expression

BackgroundThe present study was conducted to address whether the intervertebral disc of rabbit could be considered (i) as a valuable model to provide new insights into the tissue and cellular changes of Nucleus pulposus aging and (ii) as an appropriate tool to investigate the efficacy of Nucleus pulposus cell-based biotherapies.MethodsLumbar intervertebral disc from rabbits with increasing ages (1, 6 and 30 month-old) were compared by MRI and histological observation using Pfirrmann's grading and Boos' scoring respectively. The expression of transcripts (COL2A1, AGC1, COL1A1, MMP13, BMP2, MGP and p21) in Nucleus pulposus cells were analysed by quantitative real-time PCR.ResultsMRI analysis indicated an early age-dependent increase in the Pfirrmann's grading. Histological Boos' scoring was also increased. The analysis of transcript expression levels showed that COL2A1 and AGC1 were down-regulated as a function of age. Conversely, COL1A1, MMP-13, BMP-2, MGP and p21 were significantly up-regulated in the Nucleus pulposus cells of aged rabbit intervertebral disc.ConclusionsOur study describes the consistency of the rabbit as a model of intervertebral disc changes as a function of age by correlating tissue alteration with cellular modification measured.

Oxidative stress-associated shape transformation and membrane proteome remodeling in erythrocytes of end stage renal disease patients on hemodialysis

This study was designed to evaluate the oxidative stress status of erythrocytes and its association with cellular ultrastructure and membrane proteome modifications in patients with end stage renal disease (ESRD) on hemodialysis (HD). For that purpose, we studied red blood cells' (RBCs) modifications in twelve non-diabetic ESRD patients that were responsive in erythropoietin therapy. Intracellular ROS levels were measured by fluorometry, RBCs ultra-structure was examined by electron microscopy, while the membrane proteome by electrophoresis and immunoblotting. Compared to the healthy subjects, the uremic RBCs exhibited significantly increased ROS accumulation. Dialysis partially ameliorated the basal ROS levels but triggered cellular sensitivity to exogenous oxidative stimuli. Common membrane modifications involved loss, aggregation, fragmentation and carbonylation of critical components as well as over-expression of stress markers. HD significantly contributed to membrane proteome remodeling, especially for aquaporin-1, peroxiredoxin-2 and ubiquitinated proteins. The intracellular redox status and the closely associated membrane modifications seemed to be related to membrane instability, loss of surface area through vesiculation, echinocytosis and stomatocytosis. Our data evinced a network of interactions among the uremic toxins, the RBCs membrane composition and the cellular shape modifications in ESRD, which is developed around a core of oxidative provocations and cellular responses.

Abstract 3812: Retrospective analysis of Claudin-16 (CLDN16) overexpression in ovarian cancer (OVCA): Evaluation of a biomarker candidate and a novel potential therapeutic target

Abstract Our group has previously documented the overexpression of CLDN16 in OVCA; nonetheless the role of the phenomenon in cancer development and progression remains enlightened, even thought it points to cell permeability modification and tumor increased invasion potential. The present study aims to establish the clinical relevance of CLDN16 overexpression in OVCA, as well as to elucidate its role in ovarian carcinogenesis in order to future develop OVCA CLDN16-based diagnosis and/or therapeutic strategies. We had generated an OVCA database including patients clinical follow up. CLDN16 expression in tissue array platforms has been accessed by immunohistochemistry. The studied population profile corroborates to the worldwide OVCA statistics, with a prevalence of serous adenocarcinoma (serous: 42.5%, endometrioid: 17%, mucinous: 16%, clear cell: 11.3%), in menopausal women (mean age at diagnosis, 55.9 years), at advanced staged (FIGO stage I: 7.5%, II: 8.5%, III: 41.5%, VI: 27.4%; poor differentiated: 37.7%, moderately differentiated : 22.7%, well differentiated: 2.8%). Surprisingly, patients median survival rate was 12%, considerably lower than that estimated by WHO (30%). Investigation of CLDN16 expression profile have revealed, so far, that 67.74% of the cases overexpress the protein in cytoplasm. It has been also observed that CLDN16 expression is an OVCA-specific feature, as previously observed by our group. Despite the cellular localization of CLDN16 in OVCA cells, it has been postulated by others that cellular trafficking of CLDNs in cancer cells might derive from post-translational events, and might be related to vesicle trafficking or cellular matrix modification. CLDN16 is overexpressed in all OVCA histological types. To this point, we have found no differential CLDN16 expression in OVCA regarding the tumor degree of diferentiation, tumor platin responsiveness, patients survival rate or stages. Altogether, our data suggest that CLDN cytoplasm overexpression in OVCA cells might be an early event in ovarian malignant transformation, therefore playing a central role in ovarian tumorigenesis. Considering that CLDN16 silencing has been associated exclusively to decreases in magnesium serum levels, which can be exogenously replaced, it is a potential novel OVCA therapeutic target. Even though we still have to confirm CLDN16 cytoplasm localization in OVCA cells, we aim to develop a therapeutic strategy against OVCA. One possibility is encapsulate CLDN16 monoclonal antibody in lipid IgG-NaPi-Iib-immunoconjugated formulations. Ultimately, we believe that CLDN16 is an interesting and novel target to fight OVCA, which is the leading cause of gynecological cancers-related deaths among women. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3812. doi:10.1158/1538-7445.AM2011-3812

Abstract 4913: Systematic analysis of LY30-induced sensitization of cancer cells to TRAIL

Abstract TRAIL (Tumor necrosis factor-Related Apoptosis Inducing Ligand) can selectively induce apoptosis in cancer cells expressing the TRAIL receptors. However, the problem of TRAIL resistance poses a significant therapeutic challenge. We recently demonstrated that the small molecule compound, LY30, sensitizes human cancer cells to TRAIL induced apoptosis via mechanisms that involve upregulation and/or oligomerization of the death receptors. However, the underlying biology of this phenomenon remains less well defined. Here we describe a mathematical modeling approach to tease out the molecular circuitry in the synergistic response of human cervical cancer cells (HeLa) to LY30 and TRAIL mediated apoptosis. We made use of ODE (Ordinary Differential Equation) to build two distinct pathway models for Hela cells to represent TRAIL-induced apoptosis (TRAIL Model) or LY30-induced cellular modification (LY30 Model) using the existing models and previous data sets. Moreover, in accordance with the experimental settings we integrated the LY30 Model and TRAIL Model as the CTM (Combination Treatment Model). The CTM showed that the level of simulated death correlated well with experimental observations, including the synergistic level of death from the combination treatment. However, simulation and experimental data exhibited numerous mismatches for some molecules in the pathway, such as differences in the slope and onset of the synergistic activation of caspase-8 and caspase-3. Such mismatch implies some flaw in the model and thus led us to additional measurement on the early dynamics of the species involved upstream of the apoptotic pathway. In specific, assessment of cFLIP, an inhibitor of caspase-8 activation, showed slight up-regulation very early after LY30 treatment (1 hour), followed by a significant down-regulation over prolonged exposure. By including such biological phenomenon into the CTM, we are able to improve the agreement between data and simulation while maintaining the consistency in synergistic cell death. Further model analysis revealed the fluctuation of FLIP is regulated by LY30 through a complicated ratio change between two main ROS (Reactive Oxygen Species), superoxide and hydrogen peroxide. These data provide a novel means for assessment of mechanisms for small molecule-induced sensitization of human cancer cells to TRAIL mediated apoptosis, with broader clinical implications. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4913. doi:10.1158/1538-7445.AM2011-4913

Particle stimulation dephosphorylates glutathione S-transferase pi 1 of epithelial cells

Environmental pollutant exposure is associated with adverse respiratory outcomes. The phosphorylation of enzymes activates or deactivates many cellular processes and is related to the development of lung diseases such as asthma and chronic obstructive pulmonary disease. However, little is known about protein phosphorylation of bronchial epithelial cells in response to airborne particulates. Herein, we screened differentially phosphorylated proteins in TiO 2-treated epithelial cells and validated the change in GSTP1 protein phosphorylation. Two-dimensional electrophoresis was adopted for differential display proteomics of TiO 2-treated BEAS-2B cell lysates. Phosphoproteins were screened using Pro-Q ® Diamond phosphoprotein gel stain and identified by MALDI-TOF/TOF analysis. Immunoprecipitation and immunoblotting were performed for quantitative measurement of GSTP1 phosphorylation in cell lysates. Normalized relative intensities of nine phosphorylated proteins increased after TiO 2 treatment, whereas those of 12 proteins decreased in the BEAS-2B cell lysates. From gene ontology and pathway analysis, proteins involved in signal transduction were commonly identified, followed by cytoskeletal proteins, proteins from oxidation and antioxidation pathways, proteins catalyzing reductions, and those involved in cellular process, transport, and modification. Immunoblotting with anti-GSTP1 antibody demonstrated no change in GSTP1 protein levels in the lysates of BEAS-2B cells after treatment with TiO 2 particles; blotting with anti-phosphoserine and anti-phosphotyrosine antibodies showed dose-dependent decreases in phosphoserine and phosphotyrosine proteins. Stimulation with particulates phosphorylated and dephosphorylated several proteins in epithelial cells, and serine and tyrosine protein phosphorylation of GSTP1 decreased. These data indicate that airborne particles affect the pattern of phosphorylation of proteins involved in defense or apoptosis of respiratory epithelium.

PARG is recruited to DNA damage sites through poly(ADP-ribose)- and PCNA-dependent mechanisms

Post-translational poly(ADP-ribosyl)ation has diverse essential functions in the cellular response to DNA damage as it contributes to avid DNA damage detection and assembly of the cellular repair machinery but extensive modification eventually also induces cell death. While there are 17 human poly(ADP-ribose) polymerase (PARP) genes, there is only one poly(ADP-ribose) glycohydrolase (PARG) gene encoding several PARG isoforms located in different subcellular compartments. To investigate the recruitment of PARG isoforms to DNA repair sites we locally introduced DNA damage by laser microirradiation. All PARG isoforms were recruited to DNA damage sites except for a mitochondrial localized PARG fragment. Using PARP knock out cells and PARP inhibitors, we showed that PARG recruitment was only partially dependent on PARP-1 and PAR synthesis, indicating a second, PAR-independent recruitment mechanism. We found that PARG interacts with PCNA, mapped a PCNA binding site and showed that binding to PCNA contributes to PARG recruitment to DNA damage sites. This dual recruitment mode of the only nuclear PARG via the versatile loading platform PCNA and by a PAR dependent mechanism likely contributes to the dynamic regulation of this posttranslational modification and ensures the tight control of the switch between efficient DNA repair and cell death.

HMGB factors are required for posterior digit development through integrating signaling pathway activities

The chromatin factors Hmgb1 and Hmgb2 have critical roles in cellular processes, including transcription and DNA modification. To identify the function of Hmgb genes in embryonic development, we generated double mutants of Hmgb1;Hmgb2 in mice. While double null embryos arrest at E9.5, Hmgb1(-/-) ; Hmgb2(+/-) embryos exhibit a loss of digit5, the most posterior digit, in the forelimb. We show that Hmgb1(-/-) ; Hmgb2(+/-) forelimbs have a reduced level of Shh signaling, as well as a clear downregulation of Wnt and BMP target genes in the posterior region. Moreover, we demonstrate that hmgb1 and hmgb2 in zebrafish embryos enhance Wnt signaling in a variety of tissues, and that double knockdown embryos have reduced Wnt signaling and shh expression in pectoral fin buds. Our data show that Hmgb1 and Hmgb2 function redundantly to enhance Wnt signaling in embryos, and further suggest that integrating Wnt, Shh, and BMP signaling regulates the development of digit5 in forelimbs.

Transplantation of human fetal blood stem cells in the osteogenesis imperfecta mouse leads to improvement in multiscale tissue properties

AbstractOsteogenesis imperfecta (OI or brittle bone disease) is a disorder of connective tissues caused by mutations in the collagen genes. We previously showed that intrauterine transplantation of human blood fetal stem/stromal cells in OI mice (oim) resulted in a significant reduction of bone fracture. This work examines the cellular mechanisms and mechanical bone modifications underlying these therapeutic effects, particularly examining the direct effects of donor collagen expression on bone material properties. In this study, we found an 84% reduction in femoral fractures in transplanted oim mice. Fetal blood stem/stromal cells engrafted in bones, differentiated into mature osteoblasts, expressed osteocalcin, and produced COL1a2 protein, which is absent in oim mice. The presence of normal collagen decreased hydroxyproline content in bones, altered the apatite crystal structure, increased the bone matrix stiffness, and reduced bone brittleness. In conclusion, expression of normal collagen from mature osteoblast of donor origin significantly decreased bone brittleness by improving the mechanical integrity of the bone at the molecular, tissue, and whole bone levels.

Ramucirumab (IMC-1121B): Monoclonal Antibody Inhibition of Vascular Endothelial Growth Factor Receptor-2

Angiogenesis, a well-recognized characteristic of malignancy, has been exploited more than any other pathway targeted by biologic anti-neoplastic therapies. Vascular endothelial growth factor receptor-2 (VEGFR-2) is the critical receptor involved in malignant angiogenesis with its activation inducing a number of other cellular modifications resulting in tumor growth and metastases. Ramucirumab (IMC-1121B; ImClone Systems Corporation, Branchburg, NJ) is a fully human monoclonal antibody developed to specifically inhibit VEGFR-2. Ramucirumab is currently being investigated in multiple clinical trials across a variety of tumor types. Herein, angiogenesis inhibition in cancer is reviewed and up-to-date information on the clinical development of ramucirumab is presented.

Optimizing the offloading properties of the total contact cast for plantar foot ulceration

Total contact casting is the gold standard offloading treatment for plantar foot ulceration, but the optimal technique and preferred materials are poorly defined and not readily prescribed in daily practice. We investigated in-cast pressure offloading in two types of total contact casts vs. a control condition, in patients with plantar foot ulceration. In-cast walking pressures were collected using the Novel Pedar-X system in 20 participants with a plantar foot ulcer in two types of total contact casts: a conventional total contact cast and a cushion-modified total contact cast incorporating an inlay of 6 mm slow-rebound cellular urethane and 6 mm soft cellular urethane. Casts were compared with a canvas cast shoe to establish baseline pressure values. Compared with the cast shoe, the conventional total contact cast significantly reduced peak pressure at the ulcer site by 44%, mean pressure by 47% and pressure-time integral by 37% (P<0.001), while the cushion-modified total contact cast significantly reduced peak pressure at the ulcer site by 70%, mean pressure by 60% and pressure-time integral by 69% (P<0.001). Plantar pressure across the entire foot and each region of the foot was also reduced with the conventional total contact cast compared with the cast shoe, and further reduced by the cushion-modified total contact cast (P<0.05). The offloading properties of the total contact cast can be enhanced with a 12 mm cellular urethane cushion modification. Further well-designed trials are required to understand and validate this cast technique and to demonstrate healing rates and safety in different patient populations.

Long-Lasting Effects of Human Mesenchymal Stem Cell Systemic Administration on Pain-Like Behaviors, Cellular, and Biomolecular Modifications in Neuropathic Mice

Background: Neuropathic pain (NP) is an incurable disease caused by a primary lesion in the nervous system. NP is a progressive nervous system disease that results from poorly defined neurophysiological and neurochemical changes. Its treatment is very difficult. Current available therapeutic drugs have a generalized nature, sometime acting only on the temporal pain properties rather than targeting the several mechanisms underlying the generation and propagation of pain. Methods: Using biomolecular and immunohistochemical methods, we investigated the effect of the systemic injection of human mesenchymal stem cells (hMSCs) on NP relief. We used the spared nerve injury (SNI) model of NP in the mouse. hMSCs were injected into the tail vein of the mouse. Stem cell injection was performed 4 days after sciatic nerve surgery. Neuropathic mice were monitored every 10 days starting from day 11 until 90 days after surgery. Results: hMSCs were able to reduce pain-like behaviors, such as mechanical allodynia and thermal hyperalgesia, once injected into the tail vein. An anti-nociceptive effect was detectable from day 11 post surgery (7 days post cell injection). hMSCs were mainly able to home in the spinal cord and pre-frontal cortex of neuropathic mice. Injected hMSCs reduced the protein levels of the mouse pro-inflammatory interleukin IL-1β and IL-17 and increased protein levels of the mouse anti-inflammatory interleukin IL-10, and the marker of alternatively activated macrophages CD106 in the spinal cord of SNI mice. Conclusion: As a potential mechanism of action of hMSCs in reducing pain, we suggest that they could exert their beneficial action through a restorative mechanism involving: (i) a cell-to-cell contact activation mechanism, through which spinal cord homed hMSCs are responsible for switching pro-inflammatory macrophages to anti-inflammatory macrophages; (ii) secretion of a broad spectrum of molecules to communicate with other cell types. This study could provide novel findings in MSC pre-clinical biology and their therapeutic potential in regenerative medicine.

Multidrug resistance in hydrocarbon-tolerant Gram-positive and Gram-negative bacteria

New Gram-positive and Gram-negative bacteria were isolated from Poeni oily sludge, using enrichment procedures. The six Gram-positive strains belong to Bacillus, Lysinibacillus and Rhodococcus genera. The eight Gram-negative strains belong to Shewanella, Aeromonas, Pseudomonas and Klebsiella genera. Isolated bacterial strains were tolerant to saturated (i.e., n-hexane, n-heptane, n-decane, n-pentadecane, n-hexadecane, cyclohexane), monoaromatic (i.e., benzene, toluene, styrene, xylene isomers, ethylbenzene, propylbenzene) and polyaromatic (i.e., naphthalene, 2-methylnaphthalene, fluorene) hydrocarbons, and also resistant to different antimicrobial agents (i.e., ampicillin, kanamycin, rhodamine 6G, crystal violet, malachite green, sodium dodecyl sulfate). The presence of hydrophilic antibiotics like ampicillin or kanamycin in liquid LB-Mg medium has no effects on Gram-positive and Gram-negative bacteria resistance to toxic compounds. The results indicated that Gram-negative bacteria are less sensitive to toxic compounds than Gram-positive bacteria, except one bacteria belonging to Lysinibacillus genus. There were observed cellular and molecular modifications induced by ampicillin or kanamycin to isolated bacterial strains. Gram-negative bacteria possessed between two and four catabolic genes (alkB, alkM, alkB/alkB1, todC1, xylM, PAH dioxygenase, catechol 2,3-dioxygenase), compared with Gram-positive bacteria (except one bacteria belonging to Bacillus genus) which possessed one catabolic gene (alkB/alkB1). Transporter genes (HAE1, acrAB) were detected only in Gram-negative bacteria.

Bacterial and archaeal resistance to ionizing radiation

Organisms living in extreme environments must cope with large fluctuations of temperature, high levels of radiation and/or desiccation, conditions that can induce DNA damage ranging from base modifications to DNA double-strand breaks. The bacterium Deinococcus radiodurans is known for its resistance to extremely high doses of ionizing radiation and for its ability to reconstruct a functional genome from hundreds of radiation-induced chromosomal fragments. Recently, extreme ionizing radiation resistance was also generated by directed evolution of an apparently radiation-sensitive bacterial species, Escherichia coli. Radioresistant organisms are not only found among the Eubacteria but also among the Archaea that represent the third kingdom of life. They present a set of particular features that differentiate them from the Eubacteria and eukaryotes. Moreover, Archaea are often isolated from extreme environments where they live under severe conditions of temperature, pressure, pH, salts or toxic compounds that are lethal for the large majority of living organisms. Thus, Archaea offer the opportunity to understand how cells are able to cope with such harsh conditions. Among them, the halophilic archaeon Halobacterium sp and several Pyrococcus or Thermococcus species, such as Thermococcus gammatolerans, were also shown to display high level of radiation resistance. The dispersion, in the phylogenetic tree, of radioresistant prokaryotes suggests that they have independently acquired radioresistance. Different strategies were selected during evolution including several mechanisms of radiation byproduct detoxification and subtle cellular metabolism modifications to help cells recover from radiation-induced injuries, protection of proteins against oxidation, an efficient DNA repair tool box, an original pathway of DNA double-strand break repair, a condensed nucleoid that may prevent the dispersion of the DNA fragments and specific radiation-induced proteins involved in radioresistance. Here, we compare mechanisms and discuss hypotheses suggested to contribute to radioresistance in several Archaea and Eubacteria.

Mesenchymal Stem Cells in the Aging and Osteoporotic Population

Because of their ability to self-renew and differentiate, mesenchymal stem cells (MSCs) are the in vivo source for replacing lost cells in high-turnover tissues during the life of an organism. MSCs have osteogenic potential and can be eligible for the repair and maintenance of the skeleton, thus they are very attractive for tissue engineering and regenerative medicine approaches. However, many changes in their behavior, caused by aging and bone disease, have been reported in the literature. These changes, which affect MSC self-renewal ability and differentiation potentiality, are related to cell proliferation, differentiation, cell cycle phases (depending on gene modification), and cytokine and growth factor production. This review summarizes the literature related to intrinsic and extrinsic characteristics of human bone marrow or adipose tissue MSCs during aging and osteoporosis. Although some studies reveal contrasting results, the results of this review suggest that the cellular modifications due to aging and osteoporosis should be carefully considered in relation to the use of MSCs for therapeutic application.

Spinal Facet Joint Biomechanics and Mechanotransduction in Normal, Injury and Degenerative Conditions

The facet joint is a crucial anatomic region of the spine owing to its biomechanical role in facilitating articulation of the vertebrae of the spinal column. It is a diarthrodial joint with opposing articular cartilage surfaces that provide a low friction environment and a ligamentous capsule that encloses the joint space. Together with the disc, the bilateral facet joints transfer loads and guide and constrain motions in the spine due to their geometry and mechanical function. Although a great deal of research has focused on defining the biomechanics of the spine and the form and function of the disc, the facet joint has only recently become the focus of experimental, computational and clinical studies. This mechanical behavior ensures the normal health and function of the spine during physiologic loading but can also lead to its dysfunction when the tissues of the facet joint are altered either by injury, degeneration or as a result of surgical modification of the spine. The anatomical, biomechanical and physiological characteristics of the facet joints in the cervical and lumbar spines have become the focus of increased attention recently with the advent of surgical procedures of the spine, such as disc repair and replacement, which may impact facet responses. Accordingly, this review summarizes the relevant anatomy and biomechanics of the facet joint and the individual tissues that comprise it. In order to better understand the physiological implications of tissue loading in all conditions, a review of mechanotransduction pathways in the cartilage, ligament and bone is also presented ranging from the tissue-level scale to cellular modifications. With this context, experimental studies are summarized as they relate to the most common modifications that alter the biomechanics and health of the spine-injury and degeneration. In addition, many computational and finite element models have been developed that enable more-detailed and specific investigations of the facet joint and its tissues than are provided by experimental approaches and also that expand their utility for the field of biomechanics. These are also reviewed to provide a more complete summary of the current knowledge of facet joint mechanics. Overall, the goal of this review is to present a comprehensive review of the breadth and depth of knowledge regarding the mechanical and adaptive responses of the facet joint and its tissues across a variety of relevant size scales.

Nanomaterials for Regenerative Medicine

The application of nanotechnology has opened a new realm of advancement in the field of regenerative medicine and has provided hope for the culmination of long-felt needs by the development of an ideal means to control the biochemical and mechanical microenvironment for successful cell delivery and tissue regeneration. Both top-down and bottom-up approaches have been widely used in the advancement of this field, be it by improvement in scaffolds for cell growth, development of new and efficient delivery devices, cellular modification and tracking applications or by development of nanodevices such as biosensors. The current review elaborates the various nanomaterials used in regenerative medicine with a special focus on the development of this field during the last 5 years and the recent advances in their aforementioned applications. Furthermore, the key issues and challenges in using nanotechnology-based approaches are highlighted with an outlook on the likely future of nano-assisted regenerative medicine.