Second Set Of Samples Research Articles

OH-functional polyesters based on malic acid: Influence of the OH-groups onto the thermal properties

Linear aliphatic polyesters having pendant hydroxyl groups were successfully synthesised by polycondensation using scandium triflate as catalyst. A first set of samples was prepared starting with hexane-, octane-, decane-, and dodecanediol and l-malic acid as hydroxyl functional dicarboxylic acid, on the one hand, and succinic acid as non-functional analogue, on the other. A second set of samples was prepared starting with different hydroxytelechelic oligo(ε-caprolactone)s, synthesised in a ring opening polymerisation (ROP) initiated by hexanediol using the same catalyst, and reacted with l-malic acid or succinic acid, respectively. The successful reaction was confirmed by nuclear magnetic resonance (NMR) spectroscopy, size exclusion chromatography (SEC), and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF-MS). The thermal properties of both types of polyesters – with and without pendant hydroxyl groups – were analysed by differential scanning calorimetry (DSC). While all polyesters based on succinic acid are highly crystalline, for the polyesters based on l-malic acid the crystallinity depends on the concentration of pendant hydroxyl groups and on the microstructure. Hence, the influence of the hydroxyl group enables to tailor the crystallinity.

Case report

There are several different methods of testing cattle for persistent infection with bovine viral diarrhea virus (PI-BVDV), including immunohistochemistry (IHC), polymerase chain reaction (PCR), virus isolation, and antigen capture enzyme-linked immunosorbent assay (ACE). The purpose of this report is to describe a case in which a pooled PCR test (28 specimens/pool) was compared for sensitivity against an ACE test (tested as single specimens). Ear notch specimens (fresh) were collected from beef calves purchased by a stocker operation. From January through March 2010, this stocker operation received 2,424 calves and took two ear-notch specimens from each calf for PI-BVDV testing. One specimen from each calf was sent to a commercial PI-BVDV testing laboratory that utilizes real-time PCR technology in specimen pools of 28 (Laboratory A), and a second specimen was sent to a commercial PI-BVDV testing laboratory that utilizes ACE (NS23) technology using non-pooled specimens (Laboratory B). Laboratory A detected four positive specimens out of the 2,424 specimens submitted (0.165% PI prevalence), while Laboratory B detected 12 positive specimens out of the 2,424 specimens submitted (0.495% PI prevalence). The 12 specimens detected by Laboratory B included the same four detected by Laboratory A, as well as eight additional positive specimens. Upon receiving these discordant results, the attending veterinarian requested that the 12 positive specimens detected by Laboratory B be re-tested using the original specimens. These original 12 ACE-positive specimens were then re-tested as single specimens at Laboratory B using a commercially available ACE test (Erns) and real-time PCR technology. The 12 original ACE-positive specimens were again positive by these different tests. Follow-up testing on the 12 calves was conducted more than 30 days after initial testing with a second set of samples from the eight surviving calves (four of the 12 calves died between the original and the second collection). The second set of samples from the eight surviving calves initially ACE-positive by Laboratory B were tested using serum, formalin-fixed ear notches, and fresh ear notches in phosphate buffered saline. All specimens were positive by gel-based reverse transcriptase PCR on serum and ear notches (non-pooled), and all eight formalin-fixed notches were also positive by IHC.

Effect of Cd(II) on the Ripening of Ferrihydrite in Alkaline Media

AbstractTo acquire a better understanding of the influence exerted by the presence of Cd2+ during the process of transforming ferrihydrite to goethite, the morphological and structural changes of several samples obtained by the addition of Cd2+ to a suspension of nascent goethite were explored, and their chemical reactivity in acid media assessed. The samples (series Gi) were obtained by adding, at different times during the synthesis process, Cd2+ ions to ferrihydrite (Fe5HO8.4H2O) formed in alkaline media. The suspensions were aged for 5 days at 70°C, and the amorphous materials were extracted using a HCl solution (series GHCl-i). The X-ray diffraction (XRD) patterns showed that a goethite-like phase was formed, and chemical analyses indicated that the Cd content, xCd, increased with the earlier addition of the Cd2+ ions to the Fe oxyhydroxide suspension. Lattice parameters and cell volume, obtained by the Rietveld simulation of XRD data, indicated an enlargement of the cell parameters of goethite in line with the Cd-for-Fe substitution. In order to determine the influence of oxalate ions on the non-extracted solids, a second set of samples was also prepared that was kept in contact with an ammonium oxalate solution for 4 h (series Gox-i). The dissolution behavior of two series of Cd goethites and of a third series, obtained from coprecipitation of Fe3+ and Cd2+ ions in alkaline media, was observed. Kinetics measurements in 4 M HCl showed that the initial dissolution rate of samples Gox-i decreased with aging time, while the opposite effect was observed for series GHCl-i. Dissolution–time curves were well described by the Kabai equation, and activation energies were calculated using the Arrhenius equation. The results indicate that the presence of Cd during the crystallization process of goethite leads to the formation of a Cd goethite with modified morphology, structural parameters, and chemical reactivity.

EVALUACIÓN DE ANTICUERPOS CONTRA EL VIRUS DE LA DIARREA VIRAL BOVINA DE UN HATO EN PROCESO DE ERRADICACIÓN DE LA ENFERMEDAD

Antibodies against bovine viral diarrhea virus (BVDV) and antigen of BVDV wereevaluated in serum samples from cattle aged 10 to 23 months (n = 148) from a dairy herdwhere the disease eradication has been carrying out since 2002. The herd is located inSanta Rita, Arequipa, Perú, and samples were collected twice on a 30-day interval.Antibodies against BVDV and BVDV antigen were analyzed by viral neutralization andcapture ELISA tests respectively. Neutralising antibodies against BVDV were detected in6.8% (10/148) of animals, and most reactors were 19 to 23 months of age. The antibodytiter ranged from 8 to >256. None persistently infected (PI) animals were found. Theincidence of infection and seroconversion in the second set of samples was nill. Theseresults suggested that the antibodies against the BVD corresponded to residual antibodiesfrom previous infections. In addition, the results indicate the BVD infection has beencontrolled.

Potential usage of fly and bottom ash from thermal power plant ”Nikola Tesla” landfill, Serbia

In Serbia, the ash from power plants has long been labelled as hazardous waste. With the adoption of the appropriate legislation this ash became secondary raw material with the potential usage. In this paper an analysis of the fly and bottom ash composition, which are disposed of in the power plant ?Nikola Tesla A? landfill, is presented. Thirty samples, divided into three sets, were analyzed for trace elements As, Ba, Be, Cd, Co, Cr, Cu, Mn, Mo, Ni, Pb, Ti, V and Zn. The first and second set of samples were taken at the depth of 0.0-0.6 m, from cassette III, at the place of waste discharge (set I) and in the centre of the cassette (set II).The third set of samples was taken from the same cassette spot but at the different depth. The estimated variations in quality within individual sets, as well as the comparison between sets I and II, were done. The repeatability of results by the depth of cassette (set III) was also analyzed. The mixture consisting of 79.4% limestone, 17% clay, 0.5% sand, 0.55% iron ore, 0.55% from steel mill waste and 2% ash from the thermal power plant "Nikola Tesla A" was adopted as the reputable mixture for cement making. For concrete making, the same cement mixture was used but with 2.1% of the same ash material added. The results showed possibility of further fly and bottom ash use as the cement and concrete material.

Impact of shape-memory programming on mechanically-driven recovery in polymers

Shape-memory polymers (SMPs) are a class of mechanically functional “smart” materials defined by their ability to change shape upon exposure to an environmental stimulus. The shape-memory effect has traditionally been activated by thermal mechanisms via heating the polymer above a transition temperature to increase chain mobility and initiate shape recovery. This study proposes a unique approach to mechanically drive recovery in SMP networks using external forces to facilitate shape change in a material with stored strain. SMP networks were synthesized from tert-butyl acrylate and poly(ethylene glycol) dimethacrylate in three network compositions. Networks were tailored to maintain a constant glass transition temperature (∼52 °C) with increasing crosslinking density, shown by rubbery modulus values of 1.2, 3.1, and 8.2 MPa. Hollow SMP cylinders were axially elongated (programmed) to stored strain levels of approximately 25%. A second set of samples was machined to match the programmed dimensions of the SMP sample set. Compression testing revealed that the compressive strength and energy required for deformation for the programmed SMP samples were on average 62% and 52% of the as-machined samples’ values, respectively. The ratios between programmed and as-machined samples’ compressive properties were independent of both crosslinking density and temperature up to the onset of glass transition. Lastly, an interference-fit test model was used to demonstrate that mechanically-driven SMPs could immediately create and maintain a stronger fixation force compared to as-machined samples and thermally-driven SMP samples. This study introduces an approach to drive shape change that mitigates the time-temperature dependence and discusses the potential of this mechanism for biomedical devices.

Automatic Identification of Defects on Eggshell Through a Multispectral Vision System

The objective of this research was to develop an off-line artificial vision system to automatically detect defective eggshells, i.e., dirty or cracked eggshells, by employing multispectral images with the final purpose of adapting the system to an on-line grading machine. In particular, this work was focused on studying the feasibility of identifying organic stains on brown eggshells (dirty eggshell) caused by blood, feathers, feces, etc., from natural stains caused by deposits of pigments on the outer layer of clean eggshells. During the analysis, a total of 384 eggs were evaluated (clean, 148; dirty, 236). Dirty samples were evaluated visually in order to classify them according to the kind of defect (blood, feathers, and white, clear or dark feces), and clean eggshells were classified on the basis of the color of the natural stains (clear or dark). For each sample, digital images were acquired by employing a charged coupled device camera endowed with 15 monochromatic filters (440–940 nm). A Matlab® function was developed in order to automate the process and analyze images with the aim to classify samples as clean or dirty. The program was constituted by three major steps: first, the research of an opportune combination of monochromatic images in order to isolate the eggshell from the background; second, the detection of the dirt stains; third, the classification of the image samples into the dirty or clean group on the basis of the geometric characteristics of the stains (area in pixel). The proposed classification algorithm was able to correctly classify nearly 98% of the samples with a very low processing time (0.05 s). The robustness of the proposed classification was observed applying an external validation to a second set of samples (n = 178), obtaining a similar percentage of correctly classified samples (97%).

Second-harmonic optical spectroscopy on split-ring-resonator arrays

Previous second-harmonic-generation experiments on metallic split-ring-resonator arrays have been performed at fixed fundamental laser center frequency. Here, we perform nonlinear optical spectroscopy on a first set of samples, revealing pronounced resonances. Furthermore, to clarify the role of higher-order split-ring resonances, we perform additional experiments on a second set of samples in which the fundamental split-ring-resonator resonance frequencies are lithographically tuned, whereas the higher-order resonances are fixed. We find that the higher-order resonances merely reabsorb the second-harmonic generation, revealing the fundamental split-ring resonance as the nonlinear source.

Adipocyte morphometric evaluation and angiogenesis in the omentum transposed to the breast: a preliminary study

OBJECTIVE:The purpose of this study was to describe the probable mechanism of the volume increase of laparoscopically harvested omentum flaps used to treat breast deformities.METHODS:A histological analysis of omentum samples was performed to study the volume increase of laparoscopically harvested omentum flaps. Samples were harvested immediately after the transposition of the omentum from the abdominal cavity to the breast region and during the second surgical procedure for breast symmetrization of eight patients submitted to the transposition of the omentum flap. Changes in the morphometric measurements of the adipocytes (perimeter, diameter, and area), microvascular density (as measured by the CD31 endothelial marker), and immunohistochemical expression of VEGF were documented.RESULTS:The increases in adipocyte size and microvascular density were statistically significant (P ≤ 0.012). The expression levels of VEGF were lower in the second set of samples when compared to the first set, but the differences were not statistically significant (P < 0.093).CONCLUSION:These results demonstrate an increase in cellular volume as measured by adipocyte perimeter, diameter, and area. Moreover, the increase in the number of vessels in the second set of samples suggests that neoangiogenesis was stimulated by the initial increase in VEGF expression levels observed in the first set of samples. The increase in VEGF expression in the flap may have been caused by adipocyte hypertrophy resulting from neoangiogenesis.

Microbial diversity of internal environment of Johann Gregor Mendel Station, Antarctica

During January and February 2011 two sets of microbiological samples were collected inside the main building of Johann Gregor Mendel station located at the James Ross Island near the coast of Antarctica. The aim was to examine the changes of microbial profile of the antarctic station environment. The first set of samples was collected from the station environment before the staff entry, i.e. after 10 months of quiescent state when nobody was present at the station. The second set of samples was collected from the same places before the Antarctic expedition staff left the station after 45 days of the stay. The cultivation of samples was focused on mesophilic bacteria. Twenty-three strains were obtained from the Set No. 1 and 27 strains were obtained from the Set No. 2. However, 8 strains from each set were not reliably identified by mass spectrometry. Altogether 13 strains of Gram-positive bacteria were identified in the Set. No. 1, while only 7 in the Set No. 2. Contrastingly, Gram-negative bacteria were much more abundant in the Set No. 2 (12 strains) than in the Set No. 1 (2 strains). Bacillus sp. was the most common Gram-positive strain (9 isolations from the first set, 2 isolations from the second set). Pantoea agglomerans was the most common Gram-negative strain (2 isolations from the first set, 7 isolations from the second set). The first experience with the microbial profile of the research station showed that we were able to detect mainly bacteria commonly present in the outer environment that could survive under extreme conditions. We did not isolate any microbes related to human colonisation except of enterococci and Escherichia vulneris. For further investigation of the station environment, it will be necessary to choose alternative way of collection and storage of samples to ensure survival of all present bacteria.

Evaluation of crystalline perfection degree of multi‐walled carbon nanotubes: correlations between thermal kinetic analysis and micro‐Raman spectroscopy

AbstractCommercially available and laboratory‐prepared multi‐walled carbon nanotubes (MWCNTs) are systematically investigated by the use of micro‐Raman spectroscopy (MRS), thermogravimetric analysis (TGA) and complementary techniques (scanning electron microscopy (SEM) and transmission electron microscopy (TEM)) with the aim of establishing a standardised post‐growth diagnostic protocol for the assessment of their overall crystalline quality. By studying a set of ‘reference’ samples, clear correlations are evidenced between the Raman graphitisation indexes (D/G, G′/G and G′/D intensity ratios) commonly adopted to describe the crystalline arrangement of nanotubes, and their reactivity towards oxygen, as measured by the apparent activation energy needed for their oxidation, inferred from the kinetic analysis in quasi‐isothermal conditions. The higher the crystalline perfection degree, the higher the energy needed for oxidising them. The efficacy of the found correlations in indirectly assessing the reactivity of nanotubes prepared under different conditions is successfully demonstrated by the use of a second set of samples. The physical meaning and range of validity of the shown correlations are further discussed. Copyright © 2010 John Wiley &amp; Sons, Ltd.

Estimating the Stability of Organic Phosphorus in Wetland Soils

The distribution of soil P among labile and nonlabile forms can be a major determinant of agricultural and natural ecosystem productivity. Determination of soil P pools is typically performed using operationally defined chemical fractionation methods. Most of the current fractionation techniques were developed for predominately mineral soils, thus they provide only limited information on organic P (Po), particularly with respect to stability. We hypothesized that the extent to which P could be extracted from organic soils, after exposure to heat, may be related to environmental recalcitrance. We investigated two thermal methods for characterizing Po stability in organic wetland soils, an autoclave‐based and a dry heat technique. Soils from two subtropical wetlands were collected to a depth of approximately 1 m. Autoclave‐extractable P was determined by subjecting soils to 128°C and 170 kPa for 90 min in an autoclave. A second set of samples was exposed to dry heat at temperatures of 160, 200, 260, 300, 360, and 550°C. The results were compared with data from a conventional chemical P fractionation scheme. Phosphorus that could be extracted using the hot water technique declined with soil depth, representing 10 to 50% of total P in surficial soils, to 5 to 10% at a depth of 60 cm. Microbial biomass P was correlated with hot water extractable P, and represented approximately 50% of the hot water extract. In the dry heat technique, increasing the extraction temperature resulted in significantly greater extraction of Po The 360°C treatment was best able to distinguish between recalcitrant and labile Po

Texture and magnetic properties of exchange bias systems

We report on the magnetic and structural properties of IrMn/CoFe exchange bias systems deposited onto a dual seed layer of NiCr(X)/Ru(5 nm), with X=2, 6, and 20 nm. Samples with the structure NiCr (Xnm)/IrMn (7 nm)/CoFe (3 nm)/Ta (10 nm) with X=2, 6, and 20 nm were prepared by dc sputtering for magnetic characterization. A second set of samples with structure NiCr (Xnm)/IrMn (10 nm) with X=2, 6, and 20 nm were deposited onto TEM grids for structural characterization by TEM. A method of manipulating of the TEM grid to allow a qualitative analysis of the in-plane texture of the samples is described and used to analyze the microstructure of these samples. The microstructure and particularly the texture are correlated with the anisotropy constant (KAF) of the antiferromagnet (AF) layer, with an optimum NiCr seed layer of 6 nm to give a maximum value of KAF of 1.2×107 ergs/cc.

Five Serum Proteins Identified Using SELDI-TOF-MS as Potential Biomarkers of Gastric Cancer

The aims of this study were to detect serum proteomic patterns in gastric cancer serum samples using Surface-enhanced Laser Desorption/ionization-Time-of-flight-Mass Spectrometry ProteinChip array technology, to screen biomarker candidates, to build diagnostic models and to evaluate their clinical significance. Serum samples from patients with gastric cancer and normal healthy control subjects (n = 125) were analysed using surface-enhanced laser desorption/ionization technology. The spectra were generated on weak cation exchange (WCX2) chips, and protein peak clustering and classification analyses were established using Ciphergen Biomarker Wizard and Biomarker Pattern software, respectively. The diagnostic models were developed and validated by discriminant analysis. In addition, the results of the surface-enhanced laser desorption/ionization model were compared with the biomarkers carcinoembryonic antigen and carbohydrate antigen 199 in a subset of samples using a microparticle enzyme immunoassay. Five protein peaks at 2046, 3179, 1817, 1725 and 1929 m/z were automatically chosen as components of the best biomarker pattern for diagnosis of gastric cancer. In addition, we identified a single protein peak at 4665 m/z, which could distinguish between stage I/II and stage III/IV gastric cancer with a specificity and sensitivity of 91.6% (11/12) and 95.4% (21/22), respectively. When this biomarker was validated in the second set of samples, the specificity and sensitivity were 91.7% (11/12) and 86.3% (19/22), respectively. The present results suggest that serum surface-enhanced laser desorption/ionization protein profiling can distinguish patients with gastric cancer, and in particular stage I/II patients, from normal subjects with a relatively high sensitivity and specificity. Surface-enhanced Laser Desorption/ionization-Time-of-flight-Mass Spectrometry is a potential new diagnostic tool for the screening of gastric cancer.

Post-mortem degradation of selected proteins in the human brain

Event Abstract Back to Event Post-mortem degradation of selected proteins in the human brain Tamás Balázsa1*, Guillaume Lourmet1, Tamás Dóczi2, Arpád Dobolyi1 and Miklós Palkovits1 1 Hungarian Academy of Sciences and Semmelweis University, Neuromorphological and Neuroendocrine Research Laboratory, Department Anatomy, Histology and Embryology, Hungary 2 University of Pécs, Faculty of Medicine, Department of Neurosurgery, Hungary The number of proteomics studies using human brain samples has been increasing in recent years. Therefore, it is important to identify possibilities and limitations of the human post-mortem material available for protein studies. The post-mortem delay is one of the unavoidable factors to consider. We investigated the effect of post-mortem delay on the preservation/degradation of six different brain proteins. We examined the protein levels in the temporal cortex after neurosurgical removal (temporal lobectomy) from patients with drug-resistant epilepsy. Samples were taken from the middle temporal gyrus and frozen 0, 5, 15, 30, 60, 180 min after removal. During the different time periods before freezing, samples were kept on room temperature covered by saline. A second set of samples were taken from the Human Brain Tissue Bank, Budapest. These neuropatologically controlled samples were dissected from persons died in acute myocardial infarctions 1 to 16 hours after death. Similarly to the neurosurgical samples, samples were taken from the middle temporal gyrus. We performed Western blot analysis with a selection of antibodies including synapsis-related proteins (Rab 3a, α-synuclein, synaptophysin), a kinase (CalmK II), a membrane protein (PLCβ1), and a protein of the cytoskeleton (β actin). The degradation varied on the particular type of proteins. Some proteins were relatively stable within the first 3 hours including actin and synaptophysin, while others showed a considerable rapid degradation including CalmK II and Rab 3a. The effect of post-mortem delay may depend on the structural differences of brain areas. Measurements for the regional distribution of the investigated 6 proteins in the brain are in process in our laboratory. FP6, BNEII, No LSHM-CT-2004-503039. Conference: IBRO International Workshop 2010, Pécs, Hungary, 21 Jan - 23 Jan, 2010. Presentation Type: Poster Presentation Topic: Abstracts Citation: Balázsa T, Lourmet G, Dóczi T, Dobolyi A and Palkovits M (2010). Post-mortem degradation of selected proteins in the human brain. Front. Neurosci. Conference Abstract: IBRO International Workshop 2010. doi: 10.3389/conf.fnins.2010.10.00194 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 03 May 2010; Published Online: 03 May 2010. * Correspondence: Tamás Balázsa, Hungarian Academy of Sciences and Semmelweis University, Neuromorphological and Neuroendocrine Research Laboratory, Department Anatomy, Histology and Embryology, Budapest, Hungary, balazsat@ana.sote.hu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Tamás Balázsa Guillaume Lourmet Tamás Dóczi Arpád Dobolyi Miklós Palkovits Google Tamás Balázsa Guillaume Lourmet Tamás Dóczi Arpád Dobolyi Miklós Palkovits Google Scholar Tamás Balázsa Guillaume Lourmet Tamás Dóczi Arpád Dobolyi Miklós Palkovits PubMed Tamás Balázsa Guillaume Lourmet Tamás Dóczi Arpád Dobolyi Miklós Palkovits Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

FLT3 ITD Signaling Profiles in AML Samples Harboring Mutations.

Abstract 1588Poster Board I-614 BackgroundMutations in the receptor tyrosine kinase (RTK) Fms-like tyrosine kinase 3 (FLT3) gene are among the most common somatic mutations in AML with FLT3 internal tandem duplications (ITDs) occurring in 20-35% of adult and 5-15% of pediatric AML. While the presence of FLT3 ITD mutation does not appear to influence outcome to induction chemotherapy, this mutation has been shown to confer a poor prognosis with significantly shorter disease free and relapse free survival. For patients with intermediate risk cytogenetically normal AML, molecular testing for FLT3 ITD has recently been incorporated into the National Comprehensive Cancer Network (NCCN) guidelines for clinical practice. However, while molecular testing can identify a subset of patients at high risk for relapse, there remains clinical heterogeneity likely due to differences in activation of signal transduction networks. ObjectivesThis study tested the ability to use single cell network profiling (SCNP), in which cells are perturbed with extracellular modulators and their response ascertained by multiparametric flow cytometry, to identify a more clinically predictive functional readout of activation state, intracellular signaling capabilities and pathway dysregulation in the context of FLT3 mutational status. MethodsModulated SCNP was performed sequentially on two independent sets of patient samples (n=32 peripheral blood and n=85 bone marrow samples respectively). 304 and 201 “node-metric” i.e. modulated read outs of dynamic elements on individual proteins in signaling pathways were measured in the two sets respectively. These were derived from pathways known to be relevant to Flt3 WT and Flt3-ITD signaling (e.g. Ras-Raf-Erk-S6, PI3K-Akt-S6, STATs), as well as in-vitro chemotherapeutic induction of apoptosis (cleaved PARP, cleaved caspases), phosphatases, drug transporters (e.g. MDR-1, ABCG2) and expression of growth factor RTKs (e.g. Flt3R, c-Kit). ResultsIn the first study, univariate analysis revealed 76 nodes out of 304 tested that distinguished FLT3 ITD from FLT3 WT patient samples (i.e. AUC of ROC >0.7; p<0.05). Analysis of false discovery rate showed this frequency to be significantly greater than the number of nodes that can be expected by chance (p=0.0009). Although several nodes were found to be correlated, many were independent of each other and represented multiple signaling pathways. Importantly, multivariate analysis showed that combinations of independently predictive nodes improved stratification over the single nodes (AUC of ROC up to 0.99) with respect to distinguishing WT and ITD FLT3 samples. Independent analysis of a second set of samples, revealed several nodes in common between the 2 studies which distinguish FLT3 ITD from WT, including etoposide/c-PARP (apoptosis), IL-27/p-STAT3, 5 (JAK/STAT pathways) and Flt3L/p-S6 (Ras/Erk/mTOR/S6 or PI3K/mTor/S6 pathways). In both sample sets, Flt3 receptor expression did not differ significantly between FLT3 ITD and FLT3 WT samples. ConclusionsPathway analysis by SCNP revealed significant differences in signaling in FLT3 ITD relative to WT AML samples across multiple pathways. We propose that a functional signature of FLT3 signaling is distinct from the existing molecular typing and may improve the ability to predict prognostic outcomes in individual AML patients. The impact of other important prognostic, molecular markers within the FLT3 context (e.g. NPM1) are currently under investigation. DisclosuresKornblau:Nodality, Inc.: Consultancy. Rosen:Nodality, Inc.: Employment, Equity Ownership. Cleary Cohen:Nodality Inc.: Employment, Equity Ownership. Gayko:Nodality, Inc.: Employment, Equity Ownership. Putta:Nodality, Inc.: Employment, Equity Ownership. Woronicz:Nodality, Inc.: Employment, Equity Ownership. Fantl:Nodality, Inc.: Employment, Equity Ownership. Cesano:Nodality, Inc.: Employment, Equity Ownership.

Forensic analysis of explosives using isotope ratio mass spectrometry (IRMS) — Preliminary study on TATP and PETN

The application of isotopic techniques to investigations requiring the provision of evidence to a Court is limited. The objective of this research was to investigate the application of light stable isotopes and isotope ratio mass spectrometry (IRMS) to solve complex forensic cases by providing a level of discrimination not achievable utilising traditional forensic techniques. Due to the current threat of organic peroxide explosives, such as triacetone triperoxide (TATP), research was undertaken to determine the potential of IRMS to differentiate samples of TATP that had been manufactured utilising different starting materials and/or manufacturing processes. In addition, due to the prevalence of pentaerythritoltetranitrate (PETN) in detonators, detonating cord, and boosters, the potential of the IRMS technique to differentiate PETN samples from different sources was also investigated. Carbon isotope values were measured in fourteen TATP samples, with three definite groups appearing in the initial sample set based on the carbon data alone. Four additional TATP samples (in a second set of samples) were distinguishable utilising the carbon and hydrogen isotopic compositions individually, and also in combination with the oxygen isotope values. The 3D plot of the carbon, oxygen and hydrogen data demonstrated the clear discrimination of the four samples of TATP. The carbon and nitrogen isotope values measured from fifteen PETN samples, allowed samples from different sources to be readily discriminated. This paper demonstrates the successful application of IRMS to the analysis of explosives of forensic interest to assist in discriminating samples from different sources. This research represents a preliminary evaluation of the IRMS technique for the measurement of stable isotope values in TATP and PETN samples, and supports the dedication of resources for a full evaluation of this application in order to achieve Court reportable IRMS results.

Measuring Soil Nitrogen Mineralization under Field Conditions

Land application of animal manure is known to alter rates of nitrogen (N) mineralization in soils, but quantitative information concerning intensity and duration of these effects has been difficult to obtain under field conditions. We estimated net effects of manure on N mineralization in soils under field conditions in a completely randomized design, at six field sites, by comparing liquid swine (Sus scrofa) manure treatments to plots receiving no manure. Soil samples were collected immediately after manure application to determine inorganic N concentrations, and those samples were also incubated 28 d in the laboratory to determine amounts of N mineralized from the soil. Analyses and incubation were repeated on a second set of samples collected after various times, depending on the site. Differences in inorganic N concentrations were significant among treatments at all six locations for the first sampling and five of the six locations for the second sampling. In comparison, significant differences in inorganic N concentrations measured after 28 d of laboratory incubation were observed for only two of the six sites for each sampling time. Our results illustrate how to distinguish between the effects manure has on rates of N mineralization in soils and rates at which manure N is mineralized.

Identification of Metastasis-Related Proteins and Their Clinical Relevance to Triple-Negative Human Breast Cancer

Currently, there are no definite biomarkers of triple-negative breast cancer. The study aims to identify the metastasis-associated proteins of triple-negative breast tumors. A murine metastatic breast cancer model has been established by using TA2 mice. Parallel proteomic analyses were done on a murine metastatic breast cancer model and its primary breast cancer using two-dimensional gel electrophoresis. The differentially expressed proteins were detected in TA2 mice developing spontaneous breast cancer and lung metastasis. Furthermore, their expression were detected in human breast cancer with or without metastasis, and their prediction values were assessed in a second set of samples. Nineteen of 36 differentially expressed proteins were identified by peptide mass fingerprinting using matrix-assisted laser-desorption ionization-time of flight-mass spectrometry. These proteins were also validated in mouse tumor tissues by immunohistochemical staining. Actin, 14-3-3, vimentin, HSP70, CK18, and moesin were up-regulated in the metastatic tumors, whereas HSP90 and tubulin were absent or down-regulated. Furthermore, 61 patients with triple-negative breast cancer and 39 patients with estrogen receptor-positive/progesterone receptor-positive breast cancer were selected for exploring the clinical relevance of these identified proteins to human breast cancer metastasis. Expression of 14-3-3 and HSP70 was significantly correlated with metastasis of human triple-negative breast cancer. Moreover, the validation study in the second set confirmed that 14-3-3, HSP70, and their combination had high sensitivities and specificities in predicting metastatic potential of triple-negative breast cancer. These tumor metastasis-associated proteins validated may be useful as biomarkers and targets for diagnosis and treatment of human triple-negative breast cancer.

Persisting High Levels of Synovial Fluid Markers after Cartilage Repair

Local attempts to repair a cartilage lesion could cause increased levels of anabolic and catabolic factors in the synovial fluid. After repair with regenerated cartilage, the homeostasis of the cartilage ideally would return to normal. In this pilot study, we first hypothesized levels of synovial fluid markers would be higher in patients with cartilage lesions than in patients with no cartilage lesions, and then we hypothesized the levels of synovial fluid markers would decrease after cartilage repair. We collected synovial fluid samples from 10 patients before autologous chondrocyte transplantation of the knee. One year later, a second set of samples was collected and arthroscopic evaluation of the repair site was performed. Fifteen patients undergoing knee arthroscopy for various symptoms but with no apparent cartilage lesions served as control subjects. We measured synovial fluid matrix metalloproteinase-3 (MMP-3) and insulinlike growth factor-I (IGF-I) concentrations with specific activity and enzyme-linked immunosorbent assays, respectively. The levels of MMP-3 and IGF-I were higher in patients having cartilage lesions than in control subjects with no cartilage lesions. One year after cartilage repair, the lesions were filled with repair tissue, but the levels of MMP-3 and IGF-I remained elevated, indicating either graft remodeling or early degeneration. Level III, prognostic study. See the Guidelines for Authors for a complete description of levels of evidence.