Clinical outcomes following regenerative therapy of non-contained intrabony defects using a deproteinized bovine bone mineral combined with either enamel matrix derivative or collagen membrane

BACKGROUND The purpose of this study is to compare clinical outcomes in the treatment of deep non-contained intrabony defects (i.e., with ≥70% 1-wall component and a residual 2- to 3-wall component in the most apical part) using deproteinized bovine bone mineral (DBBM) combined with either enamel matrix protein derivative (EMD) or collagen membrane (CM). METHODS Forty patients with multiple intrabony defects were enrolled. Only one non-contained defect per patient with an intrabony depth ≥3 mm located in the interproximal area of single- and multirooted teeth was randomly assigned to the treatment with either EMD + DBBM (test: n = 20) or CM + DBBM (control: n = 20). At baseline and after 12 months, clinical parameters including probing depth (PD) and clinical attachment level (CAL) were recorded. The primary outcome variable was the change in CAL between baseline and 12 months. RESULTS At baseline, the intrabony component of the defects amounted to 6.1 ± 1.9 mm for EMD + DBBM and 6.0 ± 1.9 mm for CM + DBBM sites (P = 0.81). The mean CAL gain at sites treated with EMD + DBBM was not statistically significantly different (P = 0.82) compared with CM + DBBM (3.8 ± 1.5 versus 3.7 ± 1.2 mm). No statistically significant difference (P = 0.62) was observed comparing the frequency of CAL gain ≥4 mm between EMD + DBBM (60%) and CM + DBBM (50%) or comparing the frequency of residual PD ≥6 mm between EMD + DBBM (5%) and CM + DBBM (15%) (P = 0.21). CONCLUSION Within the limitations of the present study, regenerative therapy using either EMD + DBBM or CM + DBBM yielded comparable clinical outcomes in deep non-contained intrabony defects after 12 months.

Regulation of hematopoietic and leukemic stem cells by the immune system.

Hematopoietic stem cells (HSCs) are rare, multipotent cells that generate via progenitor and precursor cells of all blood lineages. Similar to normal hematopoiesis, leukemia is also hierarchically organized and a subpopulation of leukemic cells, the leukemic stem cells (LSCs), is responsible for disease initiation and maintenance and gives rise to more differentiated malignant cells. Although genetically abnormal, LSCs share many characteristics with normal HSCs, including quiescence, multipotency and self-renewal. Normal HSCs reside in a specialized microenvironment in the bone marrow (BM), the so-called HSC niche that crucially regulates HSC survival and function. Many cell types including osteoblastic, perivascular, endothelial and mesenchymal cells contribute to the HSC niche. In addition, the BM functions as primary and secondary lymphoid organ and hosts various mature immune cell types, including T and B cells, dendritic cells and macrophages that contribute to the HSC niche. Signals derived from the HSC niche are necessary to regulate demand-adapted responses of HSCs and progenitor cells after BM stress or during infection. LSCs occupy similar niches and depend on signals from the BM microenvironment. However, in addition to the cell types that constitute the HSC niche during homeostasis, in leukemia the BM is infiltrated by activated leukemia-specific immune cells. Leukemic cells express different antigens that are able to activate CD4(+) and CD8(+) T cells. It is well documented that activated T cells can contribute to the control of leukemic cells and it was hoped that these cells may be able to target and eliminate the therapy-resistant LSCs. However, the actual interaction of leukemia-specific T cells with LSCs remains ill-defined. Paradoxically, many immune mechanisms that evolved to activate emergency hematopoiesis during infection may actually contribute to the expansion and differentiation of LSCs, promoting leukemia progression. In this review, we summarize mechanisms by which the immune system regulates HSCs and LSCs.

Palaeoenvironmental changes in Central Europe (NE Poland) during the last 6200 years reconstructed from a high-resolution multi-proxy peat archive

Peat deposits from an ombrotrophic bog (north-eastern Poland) were analysed to reconstruct peatland development and environmental changes. This paper presents reconstructions of hydrological changes and plant succession over the last 6000 years. The methods included the high-resolution analysis of plant macrofossils, pollen and testate amoebae, supported by radiocarbon dating. Three main phases were identified in the history of the bog and surrounding woodland vegetation: 4000–400 BC, 400 BC–AD 1700 and AD 1700–2011. Except for terrestrialisation and the fen-to-bog transition phase, the development of bog vegetation was mainly dependent on the climate until approximately AD 1700. The dominant taxon in Gązwa bog was Sphagnum fuscum/rubellum. Woodland development was significantly affected by human activity at several time periods. The most visible human activity, manifested by the decline of deciduous species, occurred ca. 350 BC, ca. AD 250, ca. AD 1350 and after AD 1700. These events correspond to phases of human settlement in the area. During 400–300 BC, the decline of deciduous trees, primarily Carpinus, coincided with an increase in indicators of human activity and fire frequency. At ca. AD 200, Carpinus and Tilia abundance decreased, corresponding to an increased importance of cereals (Secale and Triticum). Since ca. AD 1350, the impact of Teutonic settlement is apparent, and after AD 1700, deciduous forests largely disappeared.

p62/SQSTM1 upregulation constitutes a survival mechanism that occurs during granulocytic differentiation of acute myeloid leukemia cells.

The p62/SQSTM1 adapter protein has an important role in the regulation of several key signaling pathways and helps transport ubiquitinated proteins to the autophagosomes and proteasome for degradation. Here, we investigate the regulation and roles of p62/SQSTM1 during acute myeloid leukemia (AML) cell maturation into granulocytes. Levels of p62/SQSTM1 mRNA and protein were both significantly increased during all-trans retinoic acid (ATRA)-induced differentiation of AML cells through a mechanism that depends on NF-κB activation. We show that this response constitutes a survival mechanism that prolongs the life span of mature AML cells and mitigates the effects of accumulation of aggregated proteins that occurs during granulocytic differentiation. Interestingly, ATRA-induced p62/SQSTM1 upregulation was impaired in maturation-resistant AML cells but was reactivated when differentiation was restored in these cells. Primary blast cells of AML patients and CD34(+) progenitors exhibited significantly lower p62/SQSTM1 mRNA levels than did mature granulocytes from healthy donors. Our results demonstrate that p62/SQSTM1 expression is upregulated in mature compared with immature myeloid cells and reveal a pro-survival function of the NF-κB/SQSTM1 signaling axis during granulocytic differentiation of AML cells. These findings may help our understanding of neutrophil/granulocyte development and will guide the development of novel therapeutic strategies for refractory and relapsed AML patients with previous exposure to ATRA.

Fluorescence-encoded gold nanoparticles: library design and modulation of cellular uptake into dendritic cells.

In order to harness the unique properties of nanoparticles for novel clinical applications and to modulate their uptake into specific immune cells we designed a new library of homo- and hetero-functional fluorescence-encoded gold nanoparticles (Au-NPs) using different poly(vinyl alcohol) and poly(ethylene glycol)-based polymers for particle coating and stabilization. The encoded particles were fully characterized by UV-Vis and fluorescence spectroscopy, zeta potential and dynamic light scattering. The uptake by human monocyte derived dendritic cells in vitro was studied by confocal laser scanning microscopy and quantified by fluorescence-activated cell sorting and inductively coupled plasma atomic emission spectroscopy. We show how the chemical modification of particle surfaces, for instance by attaching fluorescent dyes, can conceal fundamental particle properties and modulate cellular uptake. In order to mask the influence of fluorescent dyes on cellular uptake while still exploiting its fluorescence for detection, we have created hetero-functionalized Au-NPs, which again show typical particle dependent cellular interactions. Our study clearly prove that the thorough characterization of nanoparticles at each modification step in the engineering process is absolutely essential and that it can be necessary to make substantial adjustments of the particles in order to obtain reliable cellular uptake data, which truly reflects particle properties.

Development of a balanced experimental–computational approach to understanding the mechanics of proximal femur fractures

The majority of people who sustain hip fractures after a fall to the side would not have been identified using current screening techniques such as areal bone mineral density. Identifying them, however, is essential so that appropriate pharmacological or lifestyle interventions can be implemented. A protocol, demonstrated on a single specimen, is introduced, comprising the following components; in vitro biofidelic drop tower testing of a proximal femur; high-speed image analysis through digital image correlation; detailed accounting of the energy present during the drop tower test; organ level finite element simulations of the drop tower test; micro level finite element simulations of critical volumes of interest in the trabecular bone. Fracture in the femoral specimen initiated in the superior part of the neck. Measured fracture load was 3760 N, compared to 4871 N predicted based on the finite element analysis. Digital image correlation showed compressive surface strains as high as 7.1% prior to fracture. Voxel level results were consistent with high-speed video data and helped identify hidden local structural weaknesses. We found using a drop tower test protocol that a femoral neck fracture can be created with a fall velocity and energy representative of a sideways fall from standing. Additionally, we found that the nested explicit finite element method used allowed us to identify local structural weaknesses associated with femur fracture initiation.

Umwelt und Nutzungsgeschichte im Spiegel der Paläoökologie

Seit Jahrtausenden veränderten Menschen ihre Umwelt, direkt, etwa durch landwirtschaftliche Tätigkeiten, oder indirekt, durch Veränderungen der atmosphärischen Zusammensetzung und der biogeochemischen Zyklen. Der andauernde menschliche Einfluss führte in vielen Landökosystemen zur Entwicklung komplexer Verbindungen zwischen Mensch und Natur. Um die Rolle des Menschen und des Klimas auf Landökosysteme zu verstehen und zu entwirren, sind paläoökologische Untersuchungen an Seesedimenten und Mooren eine vielversprechende Methode. Sie setzen aber zeitlich und taxonomisch hochaufgelöste Multiproxy-Studien (z.B. Pollen-, Sporen-, Kieselalgen-, Zuckmücken- und Holzkohleanalysen) an geschichteten Sedimenten voraus. Die Zeit von der Spätantike bis ins Mittelalter ist besonders geeignet, um die Wechselwirkungen Mensch-Klima-Umwelt zu untersuchen. Einerseits wurde die Umwelt bereits zuvor durch viele tausend Jahre menschliche Landnutzung verändert, andererseits zeichnen sich lokal bis regional deutliche Landschaftsveränderungen ab: Nach einem starken Einbruch der landwirtschaftlichen Tätigkeit in den Wirren der Völkerwanderungszeit, in der die damaligen politischen und wirtschaftlichen Systeme in West-, Mittel- und Südeuropa kollabierten, erholte sich die Landnutzung im aufblühenden Frankenreich der Merowinger und noch deutlicher der Karolinger. Im 12./13. Jh. kam es zu einer Phase intensiver Städtegründungen (Kap. 4.2). Für diese Zeit stehen auch dendroklimatische Rekonstruktionen zur Verfügung, welche Niederschlag und Temperatur entkoppeln können (Büntgen et al. 2011). Am Beispiel von paläoökologischen Resultaten (Pollen, Sporen, Holzkohle) aus dem Lauerzersee diskutieren wir die Umweltveränderungen vom 5. bis ins 14. Jh. exemplarisch. Es ist zurzeit die beste vorhandene Sequenz aus dem Schweizer Mittelland und für diese Region repräsentativ.

The Little Ice Age in Scientific Perspective: Cold Spells and Caveats

Contrary to the position taken by Kelly and Ó Gráda, a rich body of regional- to large-scale temperature reconstructions that span from the last millennium to almost the entire Holocene confirms the existence of several temperature depressions that occurred at different intensities and spatial ranges between c. 1350 and 1900, thus supporting the conception of a Little Ice Age. Nonetheless, the genuine uncertainties that continue to surround paleoclimatic study suggest that methodologies and findings are subject to further refinement.

Search for supersymmetry in events with four or more leptons in √s = 8 TeV pp collisions with the ATLAS detector

Results from a search for supersymmetry in events with four or more leptons including electrons, muons and taus are presented. The analysis uses a data sample corresponding to 20.3  fb −1 of proton-proton collisions delivered by the Large Hadron Collider at s √ =8  TeV and recorded by the ATLAS detector. Signal regions are designed to target supersymmetric scenarios that can be either enriched in or depleted of events involving the production of a Z boson. No significant deviations are observed in data from standard model predictions and results are used to set upper limits on the event yields from processes beyond the standard model. Exclusion limits at the 95% confidence level on the masses of relevant supersymmetric particles are obtained. In R -parity-violating simplified models with decays of the lightest supersymmetric particle to electrons and muons, limits of 1350 and 750 GeV are placed on gluino and chargino masses, respectively. In R -parity-conserving simplified models with heavy neutralinos decaying to a massless lightest supersymmetric particle, heavy neutralino masses up to 620 GeV are excluded. Limits are also placed on other supersymmetric scenarios.

Finite element analysis predicts experimental failure patterns in vertebral bodies loaded via intervertebral discs up to large deformation

Vertebral compression fracture is a common medical problem in osteoporotic individuals. The quantitative computed tomography (QCT)-based finite element (FE) method may be used to predict vertebral strength in vivo, but needs to be validated with experimental tests. The aim of this study was to validate a nonlinear anatomy specific QCT-based FE model by using a novel testing setup. Thirty-seven human thoracolumbar vertebral bone slices were prepared by removing cortical endplates and posterior elements. The slices were scanned with QCT and the volumetric bone mineral density (vBMD) was computed with the standard clinical approach. A novel experimental setup was designed to induce a realistic failure in the vertebral slices in vitro. Rotation of the loading plate was allowed by means of a ball joint. To minimize device compliance, the specimen deformation was measured directly on the loading plate with three sensors. A nonlinear FE model was generated from the calibrated QCT images and computed vertebral stiffness and strength were compared to those measured during the experiments. In agreement with clinical observations, most of the vertebrae underwent an anterior wedge-shape fracture. As expected, the FE method predicted both stiffness and strength better than vBMD (R2 improved from 0.27 to 0.49 and from 0.34 to 0.79, respectively). Despite the lack of fitting parameters, the linear regression of the FE prediction for strength was close to the 1:1 relation (slope and intercept close to one (0.86 kN) and to zero (0.72 kN), respectively). In conclusion, a nonlinear FE model was successfully validated through a novel experimental technique for generating wedge-shape fractures in human thoracolumbar vertebrae.

Optimal Confidence Bands for Shape-Restricted Curves

Let Y be a stochastic process on [0,1] satisfying dY(t)=n 1/2 f(t)dt+dW(t) , where n≥1 is a given scale parameter (`sample size'), W is standard Brownian motion and f is an unknown function. Utilizing suitable multiscale tests, we construct confidence bands for f with guaranteed given coverage probability, assuming that f is isotonic or convex. These confidence bands are computationally feasible and shown to be asymptotically sharp optimal in an appropriate sense.

The generation of neutrophils in the bone marrow is controlled by autophagy.

Autophagy has been demonstrated to have an essential function in several cellular hematopoietic differentiation processes, for example, the differentiation of reticulocytes. To investigate the role of autophagy in neutrophil granulopoiesis, we studied neutrophils lacking autophagy-related (Atg) 5, a gene encoding a protein essential for autophagosome formation. Using Cre-recombinase mediated gene deletion, Atg5-deficient neutrophils showed no evidence of abnormalities in morphology, granule protein content, apoptosis regulation, migration, or effector functions. In such mice, however, we observed an increased proliferation rate in the neutrophil precursor cells of the bone marrow as well as an accelerated process of neutrophil differentiation, resulting in an accumulation of mature neutrophils in the bone marrow, blood, spleen, and lymph nodes. To directly study the role of autophagy in neutrophils, we employed an in vitro model of differentiating neutrophils that allowed modulating the levels of ATG5 expression, or, alternatively, intervening pharmacologically with autophagy-regulating drugs. We could show that autophagic activity correlated inversely with the rate of neutrophil differentiation. Moreover, pharmacological inhibition of p38 MAPK or mTORC1 induced autophagy in neutrophilic precursor cells and blocked their differentiation, suggesting that autophagy is negatively controlled by the p38 MAPK-mTORC1 signaling pathway. On the other hand, we obtained no evidence for an involvement of the PI3K-AKT or ERK1/2 signaling pathways in the regulation of neutrophil differentiation. Taken together, these findings show that, in contrast to erythropoiesis, autophagy is not essential for neutrophil granulopoiesis, having instead a negative impact on the generation of neutrophils. Thus, autophagy and differentiation exhibit a reciprocal regulation by the p38-mTORC1 axis.

Histone H4 mRNA from the nematode Ascaris lumbricoides is cis-spliced and polyadenylated.

A histone H4 gene from Ascaris lumbricoides contains an intron of approx. 2040 bp. Transcripts of the gene are spliced and polyadenylated. This is the first intron-containing H4 gene described for a metazoan. Notably, H4 mRNA from another nematode, Caenorhabditis elegans, is intron-less and lacks poly A (Roberts, S.B., Emmons, S.W. and Childs, G. (1989) J. Mol. Biol. 206, 567-577).