Increased plasticity of the stiffness of melanoma cells correlates with their acquisition of metastatic properties.

The stiffness of tumor cells varies during cancer progression. In particular, metastatic carcinoma cells analyzed by Atomic Force Microscopy (AFM) appear softer than non-invasive and normal cells. Here we examined by AFM how the stiffness of melanoma cells varies during progression from non-invasive Radial Growth Phase (RGP) to invasive Vertical Growth Phase (VGP) and to metastatic tumors. We show that transformation of melanocytes to RGP and to VGP cells is characterized by decreased cell stiffness. However, further progression to metastatic melanoma is accompanied by increased cell stiffness and the acquisition of higher plasticity by tumor cells, which is manifested by their ability to greatly augment or reduce their stiffness in response to diverse adhesion conditions. We conclude that increased plasticity, rather than decreased stiffness as suggested for other tumor types, is a marker of melanoma malignancy. These findings advise caution about the potential use of AFM for melanoma diagnosis. FROM THE CLINICAL EDITOR: This study investigates the changes to cellular stiffness in metastatic melanoma cells examined via atomic force microscopy. The results demonstrate that increased plasticity is a marker of melanoma malignancy, as opposed to decreased stiffness.

Antibiotic-induced modifications of the stiffness of bacterial membranes.

In the latest years the importance of high resolution analysis of the microbial cell surface has been increasingly recognized. Indeed, in order to better understand bacterial physiology and achieve rapid diagnostic and treatment techniques, a thorough investigation of the surface modifications induced on bacteria by different environmental conditions or drugs is essential. Several instruments are nowadays available to observe at high resolution specific properties of microscopic samples. Among these, AFM can routinely study single cells in physiological conditions, measuring the mechanical properties of their membrane at a nanometric scale (force volume). Such analyses, coupled with high resolution investigation of their morphological properties, are increasingly used to characterize the state of single cells. In this work we exploit such technique to characterize bacterial systems. We have performed an analysis of the mechanical properties of bacteria (Escherichia coli) exposed to different conditions. Such measurements were performed on living bacteria, by changing in real-time the liquid environment: standard phosphate buffered saline, antibiotic (ampicillin) in PBS and growth medium. In particular we have focused on the determination of the membrane stiffness modifications induced by these solutions, in particular between stationary and replicating phases and what is the effect of the antibiotic on the bacterial structure.

Atomic Force Microscopy Stiffness Tomography on Living Arabidopsis thaliana Cells Reveals the Mechanical Properties of Surface and Deep Cell-Wall Layers during Growth.

Cell-wall mechanical properties play a key role in the growth and the protection of plants. However, little is known about genuine wall mechanical properties and their growth-related dynamics at subcellular resolution and in living cells. Here, we used atomic force microscopy (AFM) stiffness tomography to explore stiffness distribution in the cell wall of suspension-cultured Arabidopsis thaliana as a model of primary, growing cell wall. For the first time that we know of, this new imaging technique was performed on living single cells of a higher plant, permitting monitoring of the stiffness distribution in cell-wall layers as a function of the depth and its evolution during the different growth phases. The mechanical measurements were correlated with changes in the composition of the cell wall, which were revealed by Fourier-transform infrared (FTIR) spectroscopy. In the beginning and end of cell growth, the average stiffness of the cell wall was low and the wall was mechanically homogenous, whereas in the exponential growth phase, the average wall stiffness increased, with increasing heterogeneity. In this phase, the difference between the superficial and deep wall stiffness was highest. FTIR spectra revealed a relative increase in the polysaccharide/lignin content.

Structural Analysis of Aggregate Blends Using the SHRP Gyratory Compactor, MLR-95-8, 1996

The Iowa Department of Transportation (IDOT) received a Strategic Highway Research Program (SHRP) gyratory compactor in December 1994. Since then IDOT has been studying the ability of the compactor to analyze fundamental properties of aggregates such as shape, texture, and gradation by studying the volumetrics of the aggregate blends under a standard load using the SHRP gyratory compactor. This method of analyzing the volumetrics of aggregate blends is similar to SHRP's fine aggregate angularity procedure, which analyzes void levels in noncompacted aggregate blends, which in turn can be used to evaluate the texture or shape of aggregates, what SHRP refers to as angularity. Research is showing that by splitting the aggregate blend on the 2.36-mm (#8) sieve and analyzing the volumetrics or angularity of the separated blend, important fundamental properties can be determined. Most important is structure (the degree and location of aggregate interlock). In addition, analysis of the volumes of the coarse and fine portions can predict the voids in the mineral aggregate and the desired asphalt content. By predicting these properties, it can be determined whether the combined aggregate blend, when mixed with asphalt cement, will produce a mix with structural adequacy to carry the designed traffic load.

Comparison of the developmental effects of two mercury compounds on glial cells and neurons in aggregate cultures of rat telencephalon.

A three-dimensional cell culture system was used as a model to study the influence of low levels of mercury in the developing brain. Aggregating cell cultures of fetal rat telencephalon were treated for 10 days either during an early developmental period (i.e., between days 5 and 15 in vitro) or during a phase of advanced maturation (i.e., between days 25 and 35) with mercury. An inorganic (HgCl2) and an organic mercury compound (monomethylmercury chloride, MeHgCl) were examined. By monitoring changes in cell type-specific enzymes activities, the concentration-dependent toxicity of the compounds was determined. In immature cultures, a general cytotoxicity was observed at 10(-6) M for both mercury compounds. In these cultures, HgCl2 appeared somewhat more toxic than MeHgCl. However, no appreciable demethylation of MeHgCl could be detected, indicating similar toxic potencies for both mercury compounds. In highly differentiated cultures, by contrast, MeHgCl exhibited a higher toxic potency than HgCl2. In addition, at 10(-6) M, MeHgCl showed pronounced neuron-specific toxicity. Below the cytotoxic concentrations, distinct glia-specific reactions could be observed with both mercury compounds. An increase in the immunoreactivity for glial fibrillary acidic protein, typical for gliosis, could be observed at concentrations between 10(-9) M and 10(-7) M in immature cultures, and between 10(-8) M and 3 x 10(-5) M in highly differentiated cultures. A conspicuous increase in the number and clustering of GSI-B4 lectin-binding cells, indicating a microglial response, was found at concentrations between 10(-10) M and 10(-7) M. These development-dependent and cell type-specific effects may reflect the pathogenic potential of long-term exposure to subclinical doses of mercury.

Audit report of the financial statements of the governmental activities, the business type activities, the aggregate discretely presented component units, each major fund and the aggregate remaining fund information of the State of Iowa as of and for the year ended June 30, 2014

Audit report of the financial statements of the governmental activities, the business type activities, the aggregate discretely presented component units, each major fund and the aggregate remaining fund information of the State of Iowa as of and for the year ended June 30, 2014

Investigation of Rapid Thermal Analysis Procedures for Prediction of the Service Life of PCCP Carbonate Coarse Aggregate, HR-337, 1993

The major objective of this research project was to use thermal analysis techniques in conjunction with x-ray analysis methods to identify and explain chemical reactions that promote aggregate related deterioration in portland cement concrete. Twenty-two different carbonate aggregate samples were subjected to a chemical testing scheme that included: • bulk chemistry (major, minor and selected trace elements) • bulk mineralogy (minor phases concentrated by acid extraction) • solid-solution in the major carbonate phases • crystallite size determinations for the major carbonate phases • a salt treatment study to evaluate the impact of deicer salts Test results from these different studies were then compared to information that had been obtained using thermogravimetric analysis techniques. Since many of the limestones and dolomites that were used in the study had extensive field service records it was possible to correlate many of the variables with service life. The results of this study have indicated that thermogravimetric analysis can play an important role in categorizing carbonate aggregates. In fact, with modern automated thermal analysis systems it should be possible to utilize such methods on a quality control basis. Strong correlations were found between several of the variables that were monitored in this study. In fact, several of the variables exhibited significant correlations to concrete service life. When the full data set was utilized (n = 18), the significant correlations to service life can be summarized as follows ( a = 5% level): • Correlation coefficient, r, = -0.73 for premature TG loss versus service life. • Correlation coefficient, r, = 0.74 for relative crystallite size versus service life. • Correlation coefficient, r, = 0.53 for ASTM C666 durability factor versus service life. • Correlation coefficient, r, = -0.52 for acid-insoluble residue versus service life. Separation of the carbonate aggregates into their mineralogical categories (i.e., calcites and dolomites) tended to increase the correlation coefficients for some specific variables (r sometimes approached 0.90); however, the reliability of such correlations was questionable because of the small number of samples that were present in this study.

Effect of Compaction on the Aggregate Fracturing of Asphalt Concrete, MLR-86-07, 1986

Other states have elected not to adopt the 75 Blow Marshall Mix Design for their heavy traffic roads. Their reasons are that the 75 blow design cracks the larger aggregates. Therefore, a limited study was done during December, 1985, to try to determine what amount of cracking took place, and in what condition the cracks are in the mix.

Investigation of Rapid Thermal Analysis Procedures for Prediction of the Service Life of PCCP Carbonate Coarse Aggregate: Phase I, Progress Report, HR-337, 1992

The major objective of this research project is to utilize thermal analysis techniques in conjunction with x-ray analysis methods to identify and explain chemical reactions that promote aggregate related deterioration in Portland cement concrete. The first year of this project has been spent obtaining and analyzing limestone and dolomite samples that exhibit a wide range of field service performance. Most of the samples chosen for the study also had laboratory durability test information (ASTM C 666, method B) that was readily available. Preliminary test results indicate that a strong relationship exists between the average crystallite size of the limestone (calcite) specimens and their apparent decomposition temperatures as measured by thermogravimetric analysis. Also, premature weight loss in the thermogravimetric analysis tests appeared to be related to the apparent decomposition temperature of the various calcite test specimens.

How Do Aggregate Fluctuations Depend on the Network Structure of the Economy?

[spa] En este artículo, analizamos la volatilidad agregada de una economía estilizada donde los agentes estann conectados en redes. Si hay relaciones estratégicas entre las acciones de los agentes, choques idiosincráticos pueden generar fluctuaciones agregadas. Demonstramos que la volatilidad agregada depende de la estructura de redes de la economía de dos maneras. Por un lado, si hay más conexiones en la economía en su conjunto, la volatilidad agregada es más baja. Por otro lado, si las conexiones están más concentradas, la volatilidad agregada es más alta. Presentamos una aplicación de nuestras predicciones teóricas que utiliza datos de EEUU de conexiones intrasectoriales y de diversificación de las empresas.

How Do Aggregate Fluctuations Depend on the Network Structure of the Economy?

[spa] En este artículo, analizamos la volatilidad agregada de una economía estilizada donde los agentes estann conectados en redes. Si hay relaciones estratégicas entre las acciones de los agentes, choques idiosincráticos pueden generar fluctuaciones agregadas. Demonstramos que la volatilidad agregada depende de la estructura de redes de la economía de dos maneras. Por un lado, si hay más conexiones en la economía en su conjunto, la volatilidad agregada es más baja. Por otro lado, si las conexiones están más concentradas, la volatilidad agregada es más alta. Presentamos una aplicación de nuestras predicciones teóricas que utiliza datos de EEUU de conexiones intrasectoriales y de diversificación de las empresas.

Acute benefits of the microbial-derived isoflavone metabolite equol on arterial stiffness in men prospectively recruited according to equol producer phenotype: a double-blind randomized controlled trial

There is much speculation with regard to the potential cardioprotective benefits of equol, a microbial-derived metabolite of the isoflavone daidzein, which is produced in the large intestine after soy intake in 30% of Western populations. Although cross-sectional and retrospective data support favorable associations between the equol producer (EP) phenotype and cardiometabolic health, few studies have prospectively recruited EPs to confirm this association. The aim was to determine whether the acute vascular benefits of isoflavones differ according to EP phenotype and subsequently investigate the effect of providing commercially produced S-(–)equol to non-EPs. We prospectively recruited male EPs and non-EPs (n = 14/ group) at moderate cardiovascular risk into a double-blind, placebocontrolled crossover study to examine the acute effects of soy isoflavones (80-mg aglycone equivalents) on arterial stiffness [carotid-femoral pulse-wave velocity (cfPWV)], blood pressure, endothelial function (measured by using the EndoPAT 2000; Itamar Medical), and nitric oxide at baseline (0 h) and 6 and 24 h after intake. In a separate assessment, non-EPs consumed 40 mg S-(–)equol with identical vascular measurements performed 2 h after intake. After soy intake, cfPWV significantly improved in EPs at 24 h (cfPWV change from 0 h: isoflavone, 20.2 6 0.2 m/s; placebo, 0.6 6 0.2 m/s; P , 0.01), which was significantly associated with plasma equol concentrations (R = 20.36, P = 0.01). No vascular effects were observed in EPs at 6 h or in non-EPs at any time point. Similarly, no benefit of commercially produced S-(–)equol was observed in non-EPs despite mean plasma equol concentrations reaching 3.2 mmol/L. Acute soy intake improved cfPWV in EPs, equating to an 11–12% reduced risk of cardiovascular disease if sustained. However, a single dose of commercially produced equol had no cardiovascular benefits in non-EPs. These data suggest that the EP phenotype is critical in unlocking the vascular benefits of equol in men, and long-term trials should focus on confirming the implications of EP phenotype on cardiovascular health. This trial was registered at clinicaltrials.gov as NCT01530893. Am J Clin Nutr doi: 10.3945/ajcn.115.125690.

Changes in the stiffness of demineralized dentin following application of tooth whitening agents

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reaction of the lateral periodontium of dogs' teeth to contaminated and noncontaminated perforations filled with mineral trioxide aggregate

It has been shown that the mineral trioxide aggregate (MTA) used to seal lateral/furcal perforations stimulates the deposition of newly formed cementum. Nevertheless, when the site of the perforation is contaminated, the healing process might occur under less favorable conditions. This study evaluated the repair healing process of noncontaminated and contaminated lateral perforations filled with MTA and the effect of previously filling the contaminated perforations with a bactericidal agent. Thirty lateral root perforations were prepared in endodontically treated dog's teeth, thus forming 3 groups with 10 specimens each. In group 1 the perforations were immediately sealed with MTA. In group 2 the perforations were left open for 7 days and thereafter sealed with MTA. In group 3 the perforations were left open for 7 days, filled temporarily with a calcium hydroxide-based paste for 14 days, and then sealed with MTA. The animals were killed after 90 days, and the pieces were prepared for histomorphologic and histomicrobiologic evaluations. The statistical analysis showed that group 1 had significantly better repair than groups 2 (P <.05) and 3 (P <.05), which validates the superior results obtained when MTA was immediately used to seal root perforations. Groups 2 and 3 had statistically similar repair to each other (P >.05). There were a larger number of cases of complete or partial biologic seal in group 1 compared with the contaminated groups. It might be concluded that the lateral root perforations sealed with MTA after contamination presented worse repair than the noncontaminated, immediately sealed perforations. The temporary filling with a bactericidal agent (calcium hydroxide-based paste) did not improve the repair of perforations exposed to contamination, and the contaminated groups presented similar results to each other.