STAT-1 expression is regulated by IGFBP-3 in malignant glioma cells and is a strong predictor of poor survival in patients with glioblastoma Laboratory investigation

Object. Insulin-like growth factor binding proteins (IGEBPs) have been implicated in the pathogenesis of glioma. In a previous study the authors demonstrated that IGFBP-3 is a novel glioblastoma biomarker associated with poor survival. Since signal transducer and activator of transcription 1 (STAT-1) has been shown to be regulated by IGFBP-3 during chondrogenesis and is a prosurvival and radioresistant molecule in different tumors, the aim in the present study was to explore the functional significance of IGFBP-3 in malignant glioma cells, to determine if STAT-1 is indeed regulated by IGFBP-3, and to study the potential of STAT-1 as a biomarker in glioblastoma. Methods. The functional significance of IGFBP-3 was investigated using the short hairpin (sh)RNA gene knockdown approach on U251MG cells. STAT-1 regulation by IGFBP-3 was tested on U251MG and U87MG cells by shRNA gene knockdown and exogenous treatment with recombinant IGFBP-3 protein. Subsequently, the expression of STAT-1 was analyzed with real-time reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) in glioblastoma and control brain tissues. Survival analyses were done on a uniformly treated prospective cohort of adults with newly diagnosed glioblastoma (136 patients) using Kaplan-Meier and Cox regression models. Results. IGFBP-3 knockdown significantly impaired proliferation, motility, migration, and invasive capacity of U251MG cells in vitro (p < 0.005). Exogenous overexpression of IGFBP-3 in U251MG and U87MG cells demonstrated STAT-1 regulation. The mean transcript levels (by real-time RT-PCR) and the mean labeling index of STAT-1 (by IHC) were significantly higher in glioblastoma than in control brain tissues (p = 0.0239 and p < 0.001, respectively). Multivariate survival analysis revealed that STAT-1 protein expression (HR 1.015, p = 0.033, 95% CI 1.001-1.029) along with patient age (HR 1.025, p = 0.005, 95% CI 1.008-1.042) were significant predictors of shorter survival in patients with glioblastoma. Conclusions. IGFBP-3 influences tumor cell proliferation, migration, and invasion and regulates STAT-1 expression in malignant glioma cells. STAT-1 is overexpressed in human glioblastoma tissues and emerges as a novel prognostic biomarker.

The generalized Onsager model for a binary gas mixture

The Onsager model for the secondary flow field in a high-speed rotating cylinder is extended to incorporate the difference in mass of the two species in a binary gas mixture. The base flow is an isothermal solid-body rotation in which there is a balance between the radial pressure gradient and the centrifugal force density for each species. Explicit expressions for the radial variation of the pressure, mass/mole fractions, and from these the radial variation of the viscosity, thermal conductivity and diffusion coefficient, are derived, and these are used in the computation of the secondary flow. For the secondary flow, the mass, momentum and energy equations in axisymmetric coordinates are expanded in an asymptotic series in a parameter epsilon = (Delta m/m(av)), where Delta m is the difference in the molecular masses of the two species, and the average molecular mass m(av) is defined as m(av) = (rho(w1)m(1) + rho(w2)m(2))/rho(w), where rho(w1) and rho(w2) are the mass densities of the two species at the wall, and rho(w) = rho(w1) + rho(w2). The equation for the master potential and the boundary conditions are derived correct to O(epsilon(2)). The leading-order equation for the master potential contains a self-adjoint sixth-order operator in the radial direction, which is different from the generalized Onsager model (Pradhan & Kumaran, J. Fluid Mech., vol. 686, 2011, pp. 109-159), since the species mass difference is included in the computation of the density, viscosity and thermal conductivity in the base state. This is solved, subject to boundary conditions, to obtain the leading approximation for the secondary flow, followed by a solution of the diffusion equation for the leading correction to the species mole fractions. The O(epsilon) and O(epsilon(2)) equations contain inhomogeneous terms that depend on the lower-order solutions, and these are solved in a hierarchical manner to obtain the O(epsilon) and O(epsilon(2)) corrections to the master potential. A similar hierarchical procedure is used for the Carrier-Maslen model for the end-cap secondary flow. The results of the Onsager hierarchy, up to O(epsilon(2)), are compared with the results of direct simulation Monte Carlo simulations for a binary hard-sphere gas mixture for secondary flow due to a wall temperature gradient, inflow/outflow of gas along the axis, as well as mass and momentum sources in the flow. There is excellent agreement between the solutions for the secondary flow correct to O(epsilon(2)) and the simulations, to within 15 %, even at a Reynolds number as low as 100, and length/diameter ratio as low as 2, for a low stratification parameter A of 0.707, and when the secondary flow velocity is as high as 0.2 times the maximum base flow velocity, and the ratio 2 Delta m/(m(1) + m(2)) is as high as 0.5. Here, the Reynolds number Re = rho(w)Omega R-2/mu, the stratification parameter A = root m Omega R-2(2)/(2k(B)T), R and Omega are the cylinder radius and angular velocity, m is the molecular mass, rho(w) is the wall density, mu is the viscosity and T is the temperature. The leading-order solutions do capture the qualitative trends, but are not in quantitative agreement.

Matrix-Assisted Refolding, Purification and Activity Assessment Using a `Form Invariant' Assay for Matrix Metalloproteinase 2 (MMP2)

Matrix metalloproteinases expression is used as biomarker for various cancers and associated malignancies. Since these proteinases can cleave many intracellular proteins, overexpression tends to be toxic; hence, a challenge to purify them. To overcome these limitations, we designed a protocol where full length pro-MMP2 enzyme was overexpressed in E. coli as inclusion bodies and purified using 6xHis affinity chromatography under denaturing conditions. In one step, the enzyme was purified and refolded directly on the affinity matrix under redox conditions to obtain a bioactive protein. The pro-MMP2 protein was characterized by mass spectrometry, CD spectroscopy, zymography and activity analysis using a simple in-house developed `form invariant' assay, which reports the total MMP2 activity independent of its various forms. The methodology yielded higher yields of bioactive protein compared to other strategies reported till date, and we anticipate that using the protocol, other toxic proteins can also be overexpressed and purified from E. coli and subsequently refolded into active form using a one step renaturation protocol.

A bio-attuned ratiometric hydrogen sulfate ion selective receptor in aqueous solvent: structural proof of the H-bonded adduct

A new cell permeable quinazoline based receptor (1) selectively senses HSO4- ions of nanomolar region in 0.1 M HEPES buffer (ethanol-water: 1/5, v/v) at biological pH over other competitive ions through the proton transfer followed by hydrogen bond formation and subsequent anion coordination to yield the LHSO4]-LH+center dot 3H(2)O (2) ensemble, which has been crystallographically characterised to ensure the structure property relationship. This non-cytotoxic HSO4- ion selective biomarker has great potential to recognize the intercellular distribution of HSO4- ions in HeLa cells under fluorescence microscope.

Impact of diurnal forcing on intraseasonal sea surface temperature oscillations in the Bay of Bengal

The diurnal cycle is an important mode of sea surface temperature (SST) variability in tropical oceans, influencing air-sea interaction and climate variability. Upper ocean mixing mechanisms are significant at diurnal time scales controlling the intraseasonal variability (ISV) of SST. Sensitivity experiments using an Ocean General Circulation Model (OGCM) for the summer monsoon of the year 2007 show that incorporation of diurnal cycle in the model atmospheric forcings improves the SST simulation at both intraseasonal and shorter time scales in the Bay of Bengal (BoB). The increase in SST-ISV amplitudes with diurnal forcing is approximate to 0.05 degrees C in the southern bay while it is approximate to 0.02 degrees C in the northern bay. Increased intraseasonal warming with diurnal forcing results from the increase in mixed layer heat gain from insolation, due to shoaling of the daytime mixed layer. Amplified intraseasonal cooling is dominantly controlled by the strengthening of subsurface processes owing to the nocturnal deepening of mixed layer. In the southern bay, intraseasonal variability is mainly determined by the diurnal cycle in insolation, while in the northern bay, diurnal cycle in insolation and winds have comparable contributions. Temperature inversions (TI) develop in the northern bay in the absence of diurnal variability in wind stress. In the northern bay, SST-ISV amplification is not as large as that in the southern bay due to the weaker diurnal variability of mixed layer depth (MLD) limited by salinity stratification. Diurnal variability of model MLD is not sufficient to create large modifications in mixed layer heat budget and SST-ISV in the northern bay.

Mercury exposure Aasessment in fish and humans from Sundarban Mangrove Wetland of India

Present study had documented total mercury levels in six commonly consumed fish species, and performed across-sectional study on local residents to gauge their intake of fish (via dietary survey) and mercury exposure (via hair biomarker analyses). Mean total mercury content in edible composites of locally-caught fishes (topse, hilsa, mackerel, topse, sardinella, khoira) was low and ranged from 0.01 to 0.11 mu g g(-1) mercury, dry weight. In a cross-sectional study of 58 area residents, the mercury content in hair ranged from 0.25 to 1.23 mu g g(-1), with a mean of 0.65 +/- 0.23 mu g g(-1), Flair mercury level was not influenced by gender, age, or occupation. Mean number of meals consumed per week was 3.1 +/- 1.1, and all participants consumed at least one fish meal per week. When related to fish consumption, a significant positive association was found between number of fish meals consumed per week and hair mercury levels.

Predicting clinical response to anticancer drugs using an ex vivo platform that captures tumour heterogeneity

Predicting clinical response to anticancer drugs remains a major challenge in cancer treatment. Emerging reports indicate that the tumour microenvironment and heterogeneity can limit the predictive power of current biomarker-guided strategies for chemotherapy. Here we report the engineering of personalized tumour ecosystems that contextually conserve the tumour heterogeneity, and phenocopy the tumour microenvironment using tumour explants maintained in defined tumour grade-matched matrix support and autologous patient serum. The functional response of tumour ecosystems, engineered from 109 patients, to anticancer drugs, together with the corresponding clinical outcomes, is used to train a machine learning algorithm; the learned model is then applied to predict the clinical response in an independent validation group of 55 patients, where we achieve 100% sensitivity in predictions while keeping specificity in a desired high range. The tumour ecosystem and algorithm, together termed the CANScript technology, can emerge as a powerful platform for enabling personalized medicine.

Effects of instabilities and coherent structures on the performance of a thermocline based thermal energy storage

We investigate the transient dynamics of disturbances inside a thermocline based molten salt thermal energy storage (TES). Numerical simulations were conducted with four inlet flow configurations. The disturbances introduced at the inlet grow via Rayleigh Taylor instability. The formed vortical motions inside the tank propagate downstream and destroy the thermocline. The vortex-thermocline interaction upsets the stratification inside the TES. The disturbance growth rate, penetration length and vortex Reynolds number are measured. The growth of penetration length prior to the vortex-thermocline interaction is quadratic. The vortex Reynolds number of the eddy which causes thermocline breakdown increases with increase in Atwood number. The impingement of vortex on thermocline is studied. (C) 2015 Elsevier Ltd. All rights reserved.

Mass spectrometry based characterization of Hb Beckman variant in a falsely elevated HbA(1c) sample

Glycated hemoglobin (HbA(1c)) is a `gold standard' biomarker for assessing the glycemic index of an individual. HbA(1c) is formed due to nonenzymatic glycosylation at N-terminal valine residue of the P-globin chain. Cation exchange based high performance liquid chromatography (CE HPLC) is mostly used to quantify HbA(1c), in blood sample. A few genetic variants of hemoglobin and post-translationally modified variants of hemoglobin interfere with CE HPLC-based quantification,. resulting in its false positive estimation. Using mass spectrometry, we analyzed a blood sample with abnormally high HbA(1c) (52.1%) in the CE HPLC method. The observed HbA(1c) did not corroborate the blood glucose level of the patient. A mass spectrometry based bottom up proteomics approach, intact globin chain mass analysis, and chemical modification of the proteolytic peptides identified the presence of Hb Beckman, a genetic variant of hemoglobin, in the experimental sample. A similar surface area to charge ratio between HbA(1c) and Hb Beckman might have resulted in the coelution of the variant with HbA(1c) in CE HPLC. Therefore, in the screening of diabetes mellitus through the estimation of HbA(1c), it is important to look for genetic variants of hemoglobin in samples that show abnormally high glycemic index, and HbA(1c) must be estimated using an alternative method. (C) 2015 Elsevier Inc. All rights reserved.

Genome-wide analysis correlates Ayurveda Prakriti

The practice of Ayurveda, the traditional medicine of India, is based on the concept of three major constitutional types (Vata, Pitta and Kapha) defined as ``Prakriti''. To the best of our knowledge, no study has convincingly correlated genomic variations with the classification of Prakriti. In the present study, we performed genome-wide SNP (single nucleotide polymorphism) analysis (Affymetrix, 6.0) of 262 well-classified male individuals (after screening 3416 subjects) belonging to three Prakritis. We found 52 SNPs (p <= 1 x 10(-5)) were significantly different between Prakritis, without any confounding effect of stratification, after 10(6) permutations. Principal component analysis (PCA) of these SNPs classified 262 individuals into their respective groups (Vata, Pitta and Kapha) irrespective of their ancestry, which represent its power in categorization. We further validated our finding with 297 Indian population samples with known ancestry. Subsequently, we found that PGM1 correlates with phenotype of Pitta as described in the ancient text of Caraka Samhita, suggesting that the phenotypic classification of India's traditional medicine has a genetic basis; and its Prakriti-based practice in vogue for many centuries resonates with personalized medicine.

Plasmonic Lipid Bilayer Membranes for Enhanced Detection Sensitivity of Biolabeling Fluorophores

Plasmonics based sensing, using the surface plasmon resonance of metal nanoparticles, has been effectively demonstrated in various applications. Extending this methodology to cell and artificial lipid bilayer membranes is extremely beneficial in enhancing the sensitivity of the detection of binding and cellular transport of molecules across such membranes. Here, the creation of an artificial plasmonic biomembrane template is demonstrated and used to show the enhanced detection sensitivity of certain widely used biomarker molecules. The efficacy of these templates is explained in terms of the ability of the hydrophobic polymer grafted gold nanoparticles used to organize, penetrate, and fluidize the membranes. The enhancement of photoluminescence of the dye molecules used occurs over a reasonably large spectral range as compared to the plasmon resonance of gold nanoparticles. The results could, possibly, be extended to cellular membranes with relevant modifications, as well as to the detection of any other biological molecule appropriately labeled with fluorescent dye molecules, and demonstrate the versatility of these plasmonic bioinspired platforms as potential biochemical sensors.

A fluorescent molecularly-imprinted polymer gate with temperature and pH as inputs for detection of alpha-fetoprotein

In this work, we have reported a new approach on the use of stimuli-responsive molecularly imprinted polymer (MIP) for trace level sensing of alpha-fetoprotein (AFP), which is a well know cancer biomarker. The stimuli-responsive MIP is composed of three components, a thermo-responsive monomer, a pH responsive component (tyrosine derivative) and a highly fluorescent vinyl silane modified carbon dot. The synthesized AFP-imprinted polymer possesses excellent selectivity towards their template molecule and dual-stimuli responsive behavior. Along with this, the imprinted polymer was also explored as `OR' logic gate with two stimuli (pH and temperature) as inputs. However, the non-imprinted polymers did not have such `OR' gate property, which confirms the role of template binding. The imprinted polymer was also used for estimation of AFP in the concentration range of 3.96-80.0 ng mL(-1), with limit of detection (LOD) 0.42 ng mL(-1). The role of proposed sensor was successfully exploited for analysis of AFP in real human blood plasma, serum and urine sample. (C) 2015 Elsevier B.V. All rights reserved.

The hemodynamic effects of in-tandem carotid artery stenosis: implications for carotid endarterectomy.

OBJECTIVES: It remains controversial whether patients with severe disease of the internal carotid artery and a coexisting stenotic lesion downstream would benefit from a carotid endarterectomy (CEA) of the proximal lesion. The aim of this study was to simulate the hemodynamic and wall shear effects of in-tandem internal carotid artery stenosis using a computational fluid dynamic (CFD) idealized model to give insight into the possible consequences of CEA on these lesions. METHODS: A CFD model of steady viscous flow in a rigid tube with two asymmetric stenoses was introduced to simulate blood flow in arteries with multiple constrictions. The effect of varying the distance between the two stenoses, and the severity of the upstream stenosis on the pressure and wall shear stress (WSS) distributions on the second plaque, was investigated. The influence of the relative positions of the two stenoses was also assessed. RESULTS: The distance between the plaques was found to have minimal influence on the overall hemodynamic effect except for the presence of a zone of low WSS (range -20 to 30 dyne/cm2) adjacent to both lesions when the two stenoses were sufficiently close (<4 times the arterial diameter). The upstream stenosis was protective if it was larger than the downstream stenosis. The relative positions of the stenoses were found to influence the WSS but not the pressure distribution. CONCLUSIONS: The geometry and positions of the lesions need to be considered when considering the hemodynamic effects of an in-tandem stenosis. Low WSS is thought to cause endothelial dysfunction and initiate atheroma formation. The fact that there was a flow recirculation zone with low WSS in between the two stenoses may demonstrate how two closely positioned plaques may merge into one larger lesion. Decision making for CEA may need to take into account the hemodynamic situation when an in-tandem stenosis is found. CFD may aid in the risk stratification of patients with this problem.

Transient Natural Ventilation of a Room with a Distributed Heat Source

We report on an experimental and theoretical study of the transient flows which develop as a naturally ventilated room adjusts from one temperature to another. We focus on a room heated from below by a uniform heat source, with both high- and low-level ventilation openings. Depending on the initial temperature of the room relative to (i) the final equilibrium temperature and (ii) the exterior temperature, three different modes of ventilation may develop. First, if the room temperature lies between the exterior and the equilibrium temperature, the interior remains well-mixed and gradually heats up to the equilibrium temperature. Secondly, if the room is initially warmer than the equilibrium temperature, then a thermal stratification develops in which the upper layer of originally hot air is displaced upwards by a lower layer of relatively cool inflowing air. At the interface, some mixing occurs owing to the effects of penetrative convection. Thirdly, if the room is initially cooler than the exterior, then on opening the vents, the original air is displaced downwards and a layer of ambient air deepens from above. As this lower layer drains, it is eventually heated to the ambient temperature, and is then able to mix into the overlying layer of external air, and the room becomes well-mixed. For each case, we present new laboratory experiments and compare these with some new quantitative models of the transient flows. We conclude by considering the implications of our work for natural ventilation of large auditoria.

Limiting States Of Internal Wave Rays

Existing models of baroclinic tides are based upon the "traditional approximation'', i. e., neglect of the horizontal component of the Earth's rotation, leading to a well- known conclusion that no freely propagating internal waves can exist beyond the critical latitude and the wave rays are symmetric to the vertical. However, recent studies have contended that the situation may change if both the vertical and horizontal components of the Earth's rotation are taken into account. With the full account of the Coriolis force, characteristics of the internal wavefield generated by tidal flow over uneven topography are investigated. It is found that "nontraditional effects'' profoundly change not only the dynamics of internal waves but also the rate at which the barotropic tidal energy is fed into the internal wavefield. Discarding the traditional approximation, internal waves are proved to be able to generate poleward of the critical latitude, rays of which are no longer symmetric and the limiting values of ray angles become greater or less than 90 degrees, depending on the local latitude and the direction of ray. More importantly, in contrast to the predictions of models based upon the traditional approximation, a substantial conversion occurs in the situations when stratification is so weak that the buoyancy frequency is below the tidal one.