Synthesis and in vitro cytotoxic evaluation of novel diazaspiro bicyclo hydantoin derivatives in human leukemia cells: A SAR study

To study the structure activity relationship (SAR) on the cytotoxic activity and probe the structural requirement for the potent antitumor activity, a series of novel diazaspiro bicyclo hydantoin derivatives were designed and synthesized. Their structures were confirmed by H-1 NMR, LCMS and IR analyses. The antiproliferative effect of these compounds were determined against human leukemia, K562 (chronic myelogenous leukemia) and CEM (T-cell leukemia) cells using trypan blue and MTT assay, and the SAR associated with the position of N-terminal substituents in diazaspiro bicyclo hydantoin have also been discussed. It has been observed that these compounds displayed strong, moderate and weak cytotoxic activities. Interestingly, compounds having electron withdrawing groups at third and fourth position of the phenyl ring displayed selectively cytotoxic activities to both the cell lines tested with IC50 value lower than 50 mu M. In addition, the cytotoxic activities of the compounds 7(a-o) bearing the substituents at N-3 position of diazaspiro bicyclo hydantoin increases in the order alkene > ester > ether and plays an important role in determining their antitumor activities. The position and number of substituents in benzyl group attached to N-8 of diazaspiro bicyclo hydantoin nucleus interacted selectively with specific targets leading to the difference of biochemical and pharmacological effects.

Transversely isotropic infinite cylindrical shell subjected to a radial axisymmetric line load

The problem of an infinite transversely isotropic circular cylindrical shell subjected to an axisymmetric radial external line load is investigated using elasticity theory, classical shell theory and shear deformation theory. The results obtained by these methods are compared for two ratios of inner to outer shell radius and for varying degrees of anisotropy. Some typical results are given here to show the effect of anisotropy and the thickness of the shell on the distribution of stresses and displacements.

Multilevel load frequency control (a perturbational approach)

A two-level control scheme for the load frequency control of a multi-area power system utilizing certain possible beneficial aspects of interconnections is described in this paper. The problem is identified as the determination of the necessary equivalent perturbation on the control distribution matrix to provide the corrective control.

Elasticity solution for a long circular sandwich cylindrical shell subjected to axisymmetric load

An elasticity solution has been obtained for a long circular sandwich cylindrical shell subjected to axisymmetric radial ring load using Love's stress function approach. Numerical results are presented for different ratios of modulus of elasticity of the layers. The results obtained from this analysis have been compared with those obtained from sandwich shell theory due to Fulton.

Load Transfer from a Smooth Elastic Pin to a Large Sheet

The plane problem of load transfer from an elastic interference or clearance fit pin to a large elastic sheet with a perfectly smooth interface is solved. As the load on the pin is monotonically increased, the pin-hole interface is in partial contact above certain critical load in interference fit and throughout the loading range in clearance fit.Such situations result in mixed boundary-value problems with moving boundaries and the arc of contact varies nonlinearly with applied load. These problems are analyzed by an inverse technique in which the arcs of contact/separation are prescribed and the causative loads are evaluated. A direct method of analysis is adopted using biharmonic polar trigonometric stress functions and a simple collocation method for satisfying the boundary conditions. A unified analytical formulation is achieved for interference and clearance fits. The solutions for the linear problem of push fits are inherent in the unified analysis. Numerical results highlighting the effects of pin and sheet elasticity parameters are presented.

Molecular convergence of hexosamine biosynthetic pathway and ER stress leading to insulin resistance in L6 skeletal muscle cells

Augmentation of hexosamine biosynthetic pathway (HBP) and endoplasmic reticulum (ER) stress were independently related to be the underlying causes of insulin resistance. We hypothesized that there might be a molecular convergence of activated HBP and ER stress pathways leading to insulin resistance. Augmentation of HBP in L6 skeletal muscle cells either by pharmacological (glucosamine) or physiological (high-glucose) means, resulted in increased protein expression of ER chaperones (viz., Grp78, Calreticulin, and Calnexin), UDP-GlcNAc levels and impaired insulin-stimulated glucose uptake. Cells silenced for O-glycosyl transferase (OGT) showed improved insulin-stimulated glucose uptake (P < 0.05) but without any effect on ER chaperone upregulation. While cells treated with either glucosamine or high-glucose exhibited increased JNK activity, silencing of OGT resulted in inhibition of JNK and normalization of glucose uptake. Our study for the first time, demonstrates a molecular convergence of O-glycosylation processes and ER stress signals at the cross-road of insulin resistance in skeletal muscle.

The use of an acetoacetyl-CoA synthase in place of a -ketothiolase enhances poly-3-hydroxybutyrate production in sugarcane mesophyll cells

Engineering the production of polyhydroxyalkanoates (PHAs) into high biomass bioenergy crops has the potential to provide a sustainable supply of bioplastics and energy from a single plant feedstock. One of the major challenges in engineering C-4 plants for the production of poly[(R)-3-hydroxybutyrate] (PHB) is the significantly lower level of polymer produced in the chloroplasts of mesophyll (M) cells compared to bundle sheath (BS) cells, thereby limiting the full PHB yield-potential of the plant. In this study, we provide evidence that the access to substrate for PHB synthesis may limit polymer production in M chloroplasts. Production of PHB in M cells of sugarcane is significantly increased by replacing -ketothiolase, the first enzyme in the bacterial PHA pathway, with acetoacetyl-CoA synthase. This novel pathway enabled the production of PHB reaching an average of 6.3% of the dry weight of total leaf biomass, with levels ranging from 3.6 to 11.8% of the dry weight (DW) of individual leaves. These yields are more than twice the level reported in PHB-producing sugarcane containing the -ketothiolase and illustrate the importance of producing polymer in mesophyll plastids to maximize yield. The molecular weight of the polymer produced was greater than 2x10(6)Da. These results are a major step forward in engineering a high biomass C-4 grass for the commercial production of PHB.

Can the direct measurement of the load applied on the residuum of individuals with transfemoral amputation fitted with bone-anchored prosthesis help to improve the rehabilitation program?

To strive to improve the rehabilitation program of individuals with transfemoral amputation fitted with bone-anchored prosthesis based on data from direct measurements of the load applied on the residuum we first of all need to understand the load applied on the fixation. Therefore the load applied on the residuum was first directly measured during standardized activities of daily living such as straight line level walking, ascending and descending stairs and a ramp and walking around a circle. From measuring the load in standardized activities of daily living the load was also measured during different phases of the rehabilitation program such as during walking with walking aids and during load bearing exercises.[1-15] The rehabilitation program for individuals with a transfemoral amputation fitted with an OPRA implant relies on a combination of dynamic and static load bearing exercises.[16-20] This presentation will focus on the study of a set of experimental static load bearing exercises. [1] A group of eleven individuals with unilateral transfemoral amputation fitted with an OPRA implant participated in this study. The load on the implant during the static load bearing exercises was measured using a portable system including a commercial transducer embedded in a short pylon, a laptop and a customized software package. This apparatus was previously shown effective in a proof-of-concept study published by Prof. Frossard. [1-9] The analysis of the static load bearing exercises included an analysis of the reliability as well as the loading compliance. The analysis of the loading reliability showed a high reliability between the loading sessions indicating a correct repetition of the LBE by the participants. [1, 5] The analysis of the loading compliance showed a significant lack of axial compliance leading to a systematic underloading of the long axis of the implant during the proposed experimental static LBE.

Fuel cells: Problems and prospects

n recent years, fuel cell technology has advanced significantly. Field trials on certain types of fuel cells have shown promise for electrical use. This article reviews the electrochemistry, problems and prospects of fuel cell systems.

PMU measurement based dynamic load modeling using SVC devices in online environment

Online dynamic load modeling has become possible with the availability of Static Voltage Compensator (SVC) and Phasor Measurement Unit (PMU) devices. The power of the load response to the small random bounded voltage fluctuations caused from SVC can be measured by PMU for modelling purposes. The aim of this paper is to illustrate the capability of identifying an aggregated load model from high voltage substation level in the online environment. The induction motor is used as the main test subject since it contributes the majority of the dynamic loads. A test system representing simple electromechanical generator model serving dynamic loads through the transmission network is used to verify the proposed method. Also, dynamic load with multiple induction motors are modeled to achieve a better realistic load representation.

Critical Evaluation of Determining Swelling Pressure by Swell-Load Method and Constant Volume Method

For any construction activity in expansive soils, determination of swelling pressure/heave is an essential step. Though many attempts have been made to develop laboratory procedures by using the laboratory one-dimensional oedometer to determine swelling pressure of expansive soils, they are reported to yield varying results. The main reason for these variations could be heterogeneous moisture distribution of the sample over its thickness. To overcome this variation the experimental procedure should be such that the soil gets fully saturated. Attempts were made to introduce vertical sand drains in addition to the top and bottom drains. In this study five and nine vertical sand drains were introduced to experimentally find out the variations in the swell and swelling pressure. The variations in the moisture content at middle, top, and bottom of the sample in the oedometer test are also reported. It is found that swell-load method is better as compared to zero-swell method. Further, five number of vertical sand drains are found to be sufficient to obtain uniform moisture content distribution.

Epithelial-mesenchymal transition in oral squamous carcinoma cells

In epithelial-mesenchymal transition (EMT), epithelial cells acquire traits typical for mesenchymal cells, dissociate their cell-cell junctions and gain the ability to migrate. EMT is essential during embryogenesis, but may also mediate cancer progression. Basement membranes are sheets of extracellular matrix that support epithelial cells. They have a major role in maintaining the epithelial phenotype and, in cancer, preventing cell migration, invasion and metastasis. Laminins are the main components of basement membranes and may actively contribute to malignancy. We first evaluated the differences between cell lines obtained from oral squamous cell carcinoma and its recurrence. As the results indicated a change from epithelial to fibroblastoid morphology, E-cadherin to N-cadherin switch, and change in expression of cytokeratins to vimentin intermediate filaments, we concluded that these cells had undergone EMT. We further induced EMT in primary tumour cells to gain knowledge of the effects of transcription factor Snail in this cell model. The E-cadherin repressors responsible for the EMT in these cells were ZEB-1, ZEB-2 and Snail, and ectopic expression of Snail was able to augment the levels of ZEB-1 and ZEB-2. We produced and characterized two monoclonal antibodies that specifically recognized Snail in cell lines and patient samples. By immunohistochemistry, Snail protein was found in mesenchymal tissues during mouse embryonal development, in fibroblastoid cells of healing skin wounds and in fibromatosis and sarcoma specimens. Furthermore, Snail localized to the stroma and borders of tumour cell islands in colon adenocarcinoma, and in laryngeal and cervical squamous cell carcinomas. Immunofluorescence labellings, immunoprecipitations and Northern and Western blots showed that EMT induced a progressive downregulation of laminin-332 and laminin-511 and, on the other hand, an induction of mesenchymal laminin-411. Chromatin immunoprecipitation revealed that Snail could directly bind upstream to the transcription start sites of both laminin α5 and α4 chain genes, thus regulating their expression. The levels of integrin α6β4, a receptor for laminin-332, as well as the hemidesmosomal complex proteins HD1/plectin and BP180 were downregulated in EMT-experienced cells. The expression of Lutheran glycoprotein, a specific receptor for laminin-511, was diminished, whereas the levels of integrins α6β1 and α1β1 and integrin-linked kinase were increased. In quantitative cell adhesion assays, the cells adhered potently to laminin-511 and fibronectin, but only marginally to laminin-411. Western blots and immunoprecipitations indicated that laminin-411 bound to fibronectin and could compromise cell adhesion to fibronectin in a dose-dependent manner. EMT induced a highly migratory and invasive tendency in oral squamous carcinoma cells. Actin-based adhesion and invasion structures, podosomes and invadopodia, were detected in the basal cell membranes of primary tumour and spontaneously transformed cancer cells, respectively. Immunofluorescence labellings showed marked differences in their morphology, as podosomes organized a ring structure with HD1/plectin, αII-spectrin, talin, focal adhesion kinase and pacsin 2 around the core filled with actin, cortactin, vinculin and filamin A. Invadopodia had no division between ring and core and failed to organize the ring proteins, but instead assembled tail-like, narrow actin cables that showed a talin-tensin switch. Time-lapse live-cell imaging indicated that both podosomes and invadopodia were long-lived entities, but the tails of invadopodia vigorously propelled in the cytoplasm and were occasionally released from the cell membrane. Invadopodia could also be externalized outside the cytoplasm, where they still retained the ability to degrade matrix. In 3D confocal imaging combined with in situ gelatin zymography, the podosomes of primary tumour cells were large, cylindrical structures that increased in time, whereas the invadopodia in EMT-driven cells were smaller, but more numerous and degraded the underlying matrix in significantly larger amounts. Fluorescence recovery after photobleaching revealed that the substructures of podosomes were replenished more rapidly with new molecules than those of invadopodia. Overall, our results indicate that EMT has a major effect on the transcription and synthesis of both intra- and extracellular proteins, including laminins and their receptors, and on the structure and dynamics of oral squamous carcinoma cells.

Androgen receptor in transcriptional regulation and carcinogenesis

Androgens control a variety of developmental processes that create the male phenotype and are important for maintaining male fertility and normal functions of tissues and organs that are not directly involved in procreation. Androgen receptor (AR) that mediates the biological actions of androgens is a member of the nuclear receptor superfamily of ligand-inducible transcription factors. Although AR was cloned over 15 years ago, the mechanisms by which it regulates gene expression are not well understood. A growing body of in vitro experimental evidence suggests that a complex network of proteins is involved in the androgen-dependent transcriptional regulation. However, the process of AR-dependent transcriptional regulation under physiological conditions is largely elusive. In the present study, a series of experiments were performed, including quantitative chromatin immunoprecipitation (ChIP) assays, to investigate AR-mediated transcription process using living prostate cancer cells. Our results show that the loading of AR and recruitment of coactivators and RNA polymerase II (Pol II) to both the promoter and enhancer of AR target genes are a transient and cyclic event that in addition to hyperacetylation, also involves dynamic changes in methylation, phosphorylation of core histone H3 in androgen-treated LNCaP cells. The dynamics of testosterone (T)-induced loading of AR onto the proximal promoters of the genes clearly differed from that loaded onto the distal enhancers. Significantly, more holo-AR was loaded onto the enhancers than the promoters, but the principal Pol II transcription complex was assembled on the promoters. By contrast, the pure antiandrogen bicalutamide (CDX) complexed to AR elicited occupancy of the PSA promoter, but was unable to load onto the PSA enhancer and was incapable of recruiting Pol II, coactivators and following changes of covalent histone modifications. The partial antagonist cyproterone acetate (CPA) and mifepristone (RU486) were capable of promoting AR loading onto both the PSA promoter and enhancer at a comparable efficiency with androgen in LNCaP cells expressing mutant AR. However, CPA- and RU486-bound AR not only recruited Pol II and coactivator p300 and GRIP1 onto the promoter and enhancer, but also recruited the corepressor NCoR onto the promoter as efficiently as CDX. In addition, we demonstrate that both proteasome and protein kinases are implicated in AR-mediated transcription. Even though proteasome inhibitor MG132 and protein kinase inhibitor DRB (5, 6-Dichlorobenzimidazole riboside) can block ligand-dependent accumulation of PSA mRNA with same efficiency, their use results in different molecular profiles in terms of the formation of AR-mediated transcriptional complex. Collectively, these results indicate that transcriptional activation by AR is a complicated process, which includes transient loading of holo-AR and recruitment of Pol II and coregulators accompanied by a cascade of distinct covalent histone modifications; This process involves both the promoter and enhancer elements, as well as other general components of the cell machineries e.g. proteasome and protein kinase; The pure antiandrogen CDX and the partial antagonist CPA and RU486 exhibit clearly different profiles in terms of their ability to induce the formation of AR-dependent transcriptional complexes and the histone modifications associated with the target genes in human prostate cancer cells. Finally, by using quantitative RT-PCR to compare the expression of sixteen AR co-regulators in prostate cancer cell lines, xenografts, and clinical prostate cancer specimens we suggest that AR co-regulators protein inhibitor of activated STAT1 (PIAS1) and steroid receptor coactivator 1(SRC1) could be involved in the progression of prostate cancer.

A coupled SPH-DEM approach to model the interactions between multiple red blood cells in motion in capillaries

Red blood cells (RBCs) are the most common type of blood cells in the blood and 99% of the blood cells are RBCs. During the circulation of blood in the cardiovascular network, RBCs squeeze through the tiny blood vessels (capillaries). They exhibit various types of motions and deformed shapes, when flowing through these capillaries with diameters varying between 5 10 µm. RBCs occupy about 45 % of the whole blood volume and the interaction between the RBCs directly influences on the motion and the deformation of the RBCs. However, most of the previous numerical studies have explored the motion and deformation of a single RBC when the interaction between RBCs has been neglected. In this study, motion and deformation of two 2D (two-dimensional) RBCs in capillaries are comprehensively explored using a coupled smoothed particle hydrodynamics (SPH) and discrete element method (DEM) model. In order to clearly model the interactions between RBCs, only two RBCs are considered in this study even though blood with RBCs is continuously flowing through the blood vessels. A spring network based on the DEM is employed to model the viscoelastic membrane of the RBC while the inside and outside fluid of RBC is modelled by SPH. The effect of the initial distance between two RBCs, membrane bending stiffness (Kb) of one RBC and undeformed diameter of one RBC on the motion and deformation of both RBCs in a uniform capillary is studied. Finally, the deformation behavior of two RBCs in a stenosed capillary is also examined. Simulation results reveal that the interaction between RBCs has significant influence on their motion and deformation.

Analysis of an orthotropic cylindrical shell having a transversely isotropic core subjected to axisymmetric load

A long two-layered circular cylinder having a thin orthotropic outer shell and a thick transversely isotropic core subjected to an axisymmetric radialv line load has been analysed. For analysis of the outer shell the classical thin shell theory was adopted and for analysis of the inner core the elasticity theory was used. The continuity of stresses and deformations at the interface has been satisfied by assumming perfect adhesion between the layers. Numerical results have been presented for two different ratios of outer shell thickness to inner radius and for three different ratios of modulus of elasticity in the radial direction of outer shell to inner core. The results have been compared with the elasticity solution of the same problem to bring out the reliability of this hybrid method. References