A novel biocompatible conducting polyvinyl alcohol (PVA)-polyvinylpyrrolidone (PVP)-hydroxyapatite (HAP) composite scaffolds for probable biological application.

We have prepared biocompatible composites of 80wt% polyvinyl alcohol (PVA)-(20wt%) polyvinylpyrrolidone (PVP) blend with different concentrations of bioactive nanohydroxyapatite, Ca10(PO4)6(HO)2 (HAP). The composite films demonstrated maximum effective conductivity (σ∼1.64×10(-4)S/m) and effective dielectric constant (ε∼290) at percolation threshold concentration (∼10wt% HAP) at room temperature. These values of σ and ε are much higher than those of PVA, PVP or HAP. Our preliminary observation indicated excellent biocompatibility of the electrospun fibrous meshes of two of these composites with different HAP contents (8.5 and 5wt% within percolation threshold concentration) using NIH 3T3 fibroblast cell line. Cells viability on the well characterized composite fibrous scaffolds was determined by MTT [3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay analysis. Enhancement of σ, due to HAP addition, was found to show increased biocompatibility of the fibrous scaffold. Enhanced σ value of the PVA/PVP-HAP composite provided supporting cues for the increased cell viability and biocompatibility of the composite fibrous meshes. Excellent biocompatibility these electrospun composite scaffolds made them to plausible potential candidates for tissue engineering or other biomedical applications.

Antitumour potential of BPT: a dual inhibitor of cdk4 and tubulin polymerization.

The marine natural product fascaplysin (1) is a potent Cdk4 (cyclin-dependent kinase 4)-specific inhibitor, but is toxic to all cell types possibly because of its DNA-intercalating properties. Through the design and synthesis of numerous fascaplysin analogues, we intended to identify inhibitors of cancer cell growth with good therapeutic window with respect to normal cells. Among various non-planar tryptoline analogues prepared, N-(biphenyl-2-yl) tryptoline (BPT, 6) was identified as a potent inhibitor of cancer cell growth and free from DNA-binding properties owing to its non-planar structure. This compound was tested in over 60 protein kinase assays. It displayed inhibition of Cdk4-cyclin D1 enzyme in vitro far more potently than many other kinases including Cdk family members. Although it blocks growth of cancer cells deficient in the mitotic-spindle checkpoint at the G0/G1 phase of the cell cycle, the block occurs primarily at the G2/M phase. BPT inhibits tubulin polymerization in vitro and acts as an enhancer of tubulin depolymerization of paclitaxel-stabilized tubulin in live cells. Western blot analyses indicated that, in p53-positive cells, BPT upregulates the expression of p53, p21 and p27 proteins, whereas it downregulates the expression of cyclin B1 and Cdk1. BPT selectively kills SV40-transformed mouse embryonic hepatic cells and human fibroblasts rather than untransformed cells. BPT inhibited the growth of several human cancer cells with an IC50<1 µM. The pharmacokinetic study in BALB/c mice indicated good plasma exposure after intravenous administration. It was found to be efficacious at 1/10th the maximum-tolerated dose (1000 mg/kg) against human tumours derived from HCT-116 (colon) and NCI-H460 (lung) cells in SCID (severe-combined immunodeficient) mice models. BPT is a relatively better anticancer agent than fascaplysin with an unusual ability to block two overlapping yet crucial phases of the cell cycle, mitosis and G0/G1. Its ability to effectively halt tumour growth in human tumour-bearing mice would suggest that BPT has the potential to be a candidate for further clinical development.

Upregulation of the microRNA cluster at the Dlk1-Dio3 locus in lung adenocarcinoma.

Mice in which lung epithelial cells can be induced to express an oncogenic Kras(G12D) develop lung adenocarcinomas in a manner analogous to humans. A myriad of genetic changes accompany lung adenocarcinomas, many of which are poorly understood. To get a comprehensive understanding of both the transcriptional and post-transcriptional changes that accompany lung adenocarcinomas, we took an omics approach in profiling both the coding genes and the non-coding small RNAs in an induced mouse model of lung adenocarcinoma. RNAseq transcriptome analysis of Kras(G12D) tumors from F1 hybrid mice revealed features specific to tumor samples. This includes the repression of a network of GTPase-related genes (Prkg1, Gnao1 and Rgs9) in tumor samples and an enrichment of Apobec1-mediated cytosine to uridine RNA editing. Furthermore, analysis of known single-nucleotide polymorphisms revealed not only a change in expression of Cd22 but also that its expression became allele specific in tumors. The most salient finding, however, came from small RNA sequencing of the tumor samples, which revealed that a cluster of ∼53 microRNAs and mRNAs at the Dlk1-Dio3 locus on mouse chromosome 12qF1 was markedly and consistently increased in tumors. Activation of this locus occurred specifically in sorted tumor-originating cancer cells. Interestingly, the 12qF1 RNAs were repressed in cultured Kras(G12D) tumor cells but reactivated when transplanted in vivo. These microRNAs have been implicated in stem cell pleuripotency and proteins targeted by these microRNAs are involved in key pathways in cancer as well as embryogenesis. Taken together, our results strongly imply that these microRNAs represent key targets in unraveling the mechanism of lung oncogenesis.

Dielectric spectroscopy of T-shaped blue-phase-III liquid crystal.

Dielectric relaxation spectra of a liquid crystalline (LC) material showing blue-phase-III (BPIII) for a considerably large temperature regime consisting of T-shaped molecules are investigated. A low frequency relaxation mode is observed in the isotropic phase (I) as well as in BPIII of the investigating material which is attributed to the short axis rotation of the T-shaped molecules. The outcome of the temperature and dc bias field variation of dielectric strength (Δε) and relaxation frequency (ν(c)) in the vicinity of the I-BPIII transition is also discussed. The temperature dependence of ν(c) in BPIII with a minor deviation from Arrhenius activities in the fluctuation-dominated nonlinear region (FDNLR) is found to follow the unusual thermal behavior of the activation energy (E(A)). The growth of pretransitional fluctuations is found to be nonlinear in the vicinity of the I-BPIII transition. A moderate growth of transition fluctuation commences from the value of the exponent α(eff)=0.38/°C, which is obtained by an exponential variation of ν(c) with respect to temperature in BPIII. Observed dynamic phenomenon in the vicinity of the I-BPIII transition regions is explained on the basis of the Landau-de Gennes and Maier-Saupe Theories.

Automatic section thickness determination using an absolute gradient focus function.

Quantitative analysis of microstructures using computerized stereology systems is an essential tool in many disciplines of bioscience research. Section thickness determination in current nonautomated approaches requires manual location of upper and lower surfaces of tissue sections. In contrast to conventional autofocus functions that locate the optimally focused optical plane using the global maximum on a focus curve, this study identified by two sharp 'knees' on the focus curve as the transition from unfocused to focused optical planes. Analysis of 14 grey-scale focus functions showed, the thresholded absolute gradient function, was best for finding detectable bends that closely correspond to the bounding optical planes at the upper and lower tissue surfaces. Modifications to this function generated four novel functions that outperformed the original. The 'modified absolute gradient count' function outperformed all others with an average error of 0.56 µm on a test set of images similar to the training set; and, an average error of 0.39 µm on a test set comprised of images captured from a different case, that is, different staining methods on a different brain region from a different subject rat. We describe a novel algorithm that allows for automatic section thickness determination based on just out-of-focus planes, a prerequisite for fully automatic computerized stereology.

Observation of relaxor ferroelectricity and multiferroic behaviour in nanoparticles of the ferromagnetic semiconductor La(2)NiMnO(6).

We report a diffuse phase transition (extending over a finite temperature range of ∼50 K) in sol-gel derived nanoparticles (∼25 nm) of the ferromagnetic double perovskite La(2)NiMnO(6). The macroscopic polarization (P-E hysteresis loop), validity of the Vogel-Fulcher relation and high dielectric permittivity (∼9 × 10(2)) confirm relaxor ferroelectric phenomena in these magnetic nanoparticles. Compared to the corresponding bulk sample, appreciably large enhancement of the magnetocapacitive effect (MC  âˆ¼ 30%) is observed even under low magnetic field (0.5 T) around the broad relaxor dielectric peak temperature (∼220 K), which is close to the ferromagnetic transition temperature (θ(f) âˆ¼ 196 K). All of these features establish the multiferroic character of the La(2)NiMnO(6) nanoparticles. The inhomogeneities arising from chemical and valence mixing in the present La(2)NiMnO(6) nanoparticles and the inter-site, Ni/Mn-site disorder along with surface disorder of the individual nanoparticles resulting in local polar regions are attributed to the observed dielectric behaviour of the nanoparticles. The wave vector dependent spin-pair correlation is considered to be the plausible cause of the colossal magnetocapacitive response near the transition temperature. High permittivity and large magnetocapacitive properties make these ferromagnetic La(2)NiMnO(6) nanoparticles technologically important.

Incidence of ESBL producers amongst Gram-negative bacilli isolated from intra-abdominal infections across India (based on SMART study, 2007 data).

OBJECTIVES: This study was conducted in 9 centers spread over India from January 1 to December 31, 2007 to monitor in vitro susceptibility of Gram-negative bacilli to Group I carbapenem, ertapenem and other antimicrobials in intra-abdominal infections and to identify early changes in susceptibility pattern of community or hospital acquired organisms, with a focus on ESBL producers. MATERIAL AND METHODS: Gram-negative bacilli isolated from intra-abdominal samples of patients with documented intra-abdominal infections were processed for identification by conventional/ automated methods and antimicrobial susceptibility by Micro-Scan (Siemens) MIC panel against 12 antimicrobials (3rd and 4th generation cephalosporins, Groups I and II carbapenems, amikacin, levofloxacin, amoxicillin-clavulanic acid and piperacillin-tazobactam). RESULTS: A total of 588 isolates were identified, of which 351 (60%) were E. coli and 114 (19%) were Klebsiella spp. 79% of E. coli and 70% of Klebsiella spp. were ESBL producers in general. 110 of E. coli and 35 of Klebsiella isolates were from community-acquired intra-abdominal infections. 80% of E. coli and 63% of Klebsiella isolates from community-acquired infections were ESBL producers, against 79% of E. coli and 73% of Klebsiella isolates from hospital-acquired infections. Amongst the ESBL-positive isolates of E. coli, 94% were susceptible in vitro to ertapenem, 96% to imipenem and 76% to piperacillin-tazobactam. For ESBL-positive isolates of Klebsiella spp., the corresponding figures were 80%, 94% and 59% respectively. CONCLUSION: The study showed a high incidence of ESBL-producers amongst Enterobacteriaceae isolates from intra-abdominal infections in both community-acquired and hospital-acquired settings across India. Ertapenem was comparable with imipenem against ESBL-positive E. coli isolates, while imipenem was more effective than ertapenem against ESBL-positive Klebsiella isolates.

Identification and characterization of amylase-binding protein C from Streptococcus mitis NS51.

A substantial proportion of the streptococcal species found in dental plaque biofilms are able to interact with the abundant salivary enzyme alpha-amylase. These streptococci produce proteins that specifically bind amylase. An important plaque species, Streptococcus mitis, secretes a 36-kDa amylase-binding protein into the extracellular milieu. Proteins precipitated from S. mitis NS51 cell culture supernatant by the addition of purified salivary amylase were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to a membrane, and a prominent 36-kDa band was cut from the membrane and sequenced to yield the N-terminal amino acid sequence DSQAQYSNGV. Searching the S. mitis genome sequence database revealed a single open reading frame containing this sequence, and the gene was amplified by the S. mitis genomic DNA polymerase chain reaction. The coding region of this open reading frame, designated amylase-binding protein C (AbpC), was cloned into an Escherichia coli expression vector and the recombinant AbpC (rAbpC) was purified from the soluble fraction of the E. coli cell lysate. Purified AbpC was found to interact with immobilized amylase, confirming AbpC as a new streptococcal amylase-binding protein.

Critical evaluation of toxic versus beneficial effects of methylglyoxal.

In various organisms, an array of enzymes is involved in the synthesis and breakdown of methylglyoxal. Through these enzymes, it is intimately linked to several other physiologically important metabolites, suggesting that methylglyoxal has some important role to play in the host organism. Several in vitro and in vivo studies showed that methylglyoxal acts specifically against different types of malignant cells. These studies culminated in a recent investigation to evaluate a methylglyoxal-based formulation in treating a small group of cancer patients, and the results were promising. Methylglyoxal acts against a number of pathogenic microorganisms. However, recent literature abounds with the toxic effects of methylglyoxal, which are supposed to be mediated through methylglyoxal-derived advanced glycation end products (AGE). Many diseases such as diabetes, cataract formation, hypertension, and uremia are proposed to be intimately linked with methylglyoxal-derived AGE. However methylglyoxal-derived AGE formation and subsequent pathogenesis might be a very minor event because AGE are nonspecific reaction products that are derived through the reactions of carbonyl groups of reducing sugars with amino groups present in the side chains of lysine and arginine and in terminal amino groups of proteins. Moreover, the results of some in vitro experiments with methylglyoxal under non-physiological conditions were extrapolated to the in vivo situation. Some experiments even showed contradictory results and were differently interpreted. For this reason conclusions about the potential beneficial effects of methylglyoxal have often been neglected, thus hindering the advancement of medical science and causing some confusion in fundamental understanding. Overall, the potential beneficial effects of methylglyoxal far outweigh its possible toxic role in vivo, and it should be utilized for the benefit of suffering humanity.

Handwritten numeral databases of Indian scripts and multistage recognition of mixed numerals.

This article primarily concerns the problem of isolated handwritten numeral recognition of major Indian scripts. The principal contributions presented here are (a) pioneering development of two databases for handwritten numerals of two most popular Indian scripts, (b) a multistage cascaded recognition scheme using wavelet based multiresolution representations and multilayer perceptron classifiers and (c) application of (b) for the recognition of mixed handwritten numerals of three Indian scripts Devanagari, Bangla and English. The present databases include respectively 22,556 and 23,392 handwritten isolated numeral samples of Devanagari and Bangla collected from real-life situations and these can be made available free of cost to researchers of other academic Institutions. In the proposed scheme, a numeral is subjected to three multilayer perceptron classifiers corresponding to three coarse-to-fine resolution levels in a cascaded manner. If rejection occurred even at the highest resolution, another multilayer perceptron is used as the final attempt to recognize the input numeral by combining the outputs of three classifiers of the previous stages. This scheme has been extended to the situation when the script of a document is not known a priori or the numerals written on a document belong to different scripts. Handwritten numerals in mixed scripts are frequently found in Indian postal mails and table-form documents.

Correlation of structural, magnetic and transport properties with the tolerance factor in a low-doped La(0.875)Sr(0.125-x)Ca(x)MnO(3), (0 ≤x≤0.125) system: cross-over from Mott to Shklovskii-Efros variable range hopping conduction.

Samples of the low-doped manganite La(0.875)Sr(0.125-x)Ca(x)MnO(3) (0≤x≤0.125) have been synthesized and the effect on the structural, magnetic and transport properties of decreasing the tolerance factor by replacing larger Sr(2+) ions with smaller Ca(2+) ions are reported. For samples with x≥0.0625, a concentration (x) dependent structural transition (rhombohedral ([Formula: see text]) to orthorhombic (Pnma)) has been detected at room temperature and the Curie temperature T(C) is found to decrease with increased Ca doping level. For samples with x≤0.0625, a narrow metallic region exists and the corresponding insulator to metal transition temperature T(MI) decreases with increasing Ca content, i.e. decreasing tolerance factor. In the paramagnetic region, x dependent crossover from Mott variable range hopping (Mott-VRH) to Shklovskii-Efros variable range hopping (SE-VRH) occurs as the Ca content increases. The thermoelectric power (TEP) of the samples increases substantially, varying inversely with the tolerance factor. These results are analysed from the consideration of increased bending of the Mn-O-Mn bond with the decrease of the average ionic radius of the A-site element [Formula: see text] and the tolerance factor t, which causes narrowing of the bandwidth, decrease of mobility of e(g) electrons and weakening of the double exchange (DE) interaction associated with the substitution of Ca.

Frequency-plane analysis of normal and pathological ECG signals for disease identification.

In this paper a frequency plane analysis of both normal and diseased ECG signals is performed specifically for disease identification. Image processing techniques are used to develop an automated data acquisition package of 12 lead ECG signals from paper records. A regeneration domain is also developed to check the captured pattern with the original wave shape. A QRS complex detector with an accuracy level approximately 98.4% in up to 30% signal to noise level is developed. Discrete Fourier transform (DFT) is performed to obtain the frequency spectrum of every ECG signal. Some interesting amplitude and phase response properties of chest lead V2, V3, V4, V6 and limb lead I, II, III, AVL, AVF are seen. Both amplitude and phase properties are different for normal and diseased subjects and can serve an important role in disease identification. A statistical analysis of amplitude property is carried out to show that this property is significantly different for normal and diseased subjects.

Recognition of online handwritten mathematical expressions.

This paper aims at automatic understanding of online handwritten mathematical expressions (MEs) written on an electronic tablet. The proposed technique involves two major stages: symbol recognition and structural analysis. Combination of two different classifiers have been used to achieve high accuracy for the recognition of symbols. Several online and offline features are used in the structural analysis phase to identify the spatial relationships among symbols. A context-free grammar has been designed to convert the input expressions into their corresponding T(E)X strings which are subsequently converted into MathML format. Contextual information has been used to correct several structure interpretation errors. A new method for evaluating performance of the proposed system has been formulated. Experiments on a dataset of considerable size strongly support the feasibility of the proposed system.

Generation of digital time database from paper ECG records and Fourier transform-based analysis for disease identification.

ECG signals recorded on paper are transferred to the digital time database with the help of an automated data extraction system developed here. A flatbed scanner is used to form an image database of each 12-lead ECG signal. Those images are then fed into a Pentium PC having a system to extract pixel-to-pixel co-ordinate information to form a raw database with the help of some image processing techniques. These raw data are then ported to the regeneration domain of the system to check the captured pattern with the original wave shape. The sampling period of each ECG signal is computed after detection of QRS complex. Finally, discrete Fourier transform of the generated database is performed to observe the frequency response properties of every ECG signal. Some interesting amplitude properties of monopolar chest lead V4 and V6 are noticed which are stated.

Dielectric relaxation behavior of a liquid crystal showing an unusual type of antiferroelectric-ferroelectric-antiferroelectric phase sequence.

The temperature and frequency dependent dielectric relaxation behavior of a liquid crystalline (S)-(+)-1-methylheptyl 4-[2-(4-alkoxyphenyl) thiophene-5-carbonylthiooxy] benzoate system is reported. Interesting successive antiferroelectric-ferroelectric-antiferroelectric (AF-FE-AF) phase transitions are observed in this system resembling the successive phase transitions observed in crystalline Rochelle salt. The smectic-C* (SmC*) to AF1 phase transition (around 103.0 degrees C) is first order in nature, predicted from the use of Orihara and Ishibashi theory. It is also found that a contribution of the ferroelectric SmC* phase ordering penetrates even in the antiferroelectric AF1 (SmC(*)(A)) and AF2 (SmC(*)(A)) phases very close to the SmC*-AF1 and SmC*-AF2 phase boundaries (critical regions). It is suggested that this type of mixing of AF and FE phases might cause surface induced ferroelectric- or ferroelectric-type ordering near the AF-FE phase transitions. A soft mode with Debye-type dispersion was observed in the SmA phase. The thermal behaviors of dielectric dispersion, absorption, and dielectric strength in different phases are also reported and discussed.

Potentiality of a new compound for in vitro differentiation between halophilic and non-halophilic vibrios.

Sensitivity of 21 halophilic vibrios and 16 clinical isolates of non-halophilic vibrios was determined against a new possible antivibrio agent, a pyrimidine analogue, 4, 6-dimethylpyrimidine -2-thiol (4,6-DMPT). It appeared to be a vibriocidal agent, having a mean MIC and MBC of 32 microg/ml for halophilic strains and 64 microg/ml for non-halophilic strains and an LD50 of 300 mg/Kg body weight of mice. Thus, 4,6-DMPT may help an in vitro distinction between halophilic and non-halophilic vibrios. Sensitivity of these strains was also studied with respect to pteridine, crystal violet and Tween 80 hydrolysis as further markers distinguishing between these 2 groups which could also be differentiated by their growth on TCBS or/and CLED media.

Dielectric investigation of electrically oriented ferroelectric smectic mixture CS-1013.

From dielectric spectroscopic study, a first-order ferroelectric phase transition has been observed in ferroelectric smectic mixture CS-1013 having the phase sequence Cr-SmC*-SmA-N*-Iso. Frequency (100 Hz-10 MHz) and temperature-dependent dielectric measurements have been performed on an electrically aligned sample (thickness 15+/-1 microm) gold coated on glass plates. In the unidirectionally aligned sample, two dielectric relaxation modes (Goldstone mode and soft mode) have been clearly observed in the ferroelectric SmC* phase while only one relaxation mode (soft mode) is visualized in the paraelectric SmA phase. Low-frequency molecular relaxation was also observed in the smectic phases. The experimental results have also been analyzed at different temperatures and biasing voltages for an understanding of the dynamics of dielectric processes in the ferroelectric phase. Finally, we proposed the "pseudospin" model for understanding the ferroelectric-antiferroelectric transition in liquid crystals. We associate the tilt angle straight theta and the pitch of the helix, respectively, with biaxial (b) and uniaxial (u) anisotropy parameters as fluctuating parameters around their stability limit (corresponding to the crystalline values). Here, the director acts as the pseudospin variable. This gives rise to a transverse Ising type (or anisotropic Heisenberg model under the mean-field approximation). It is then shown that such a model with fluctuations of (b) and (u) would explain the ferroelectric and antiferroelectric phase transitions in such liquid crystals. Using Landau theory and the stability conditions, we have also shown, in brief, the feasibility of different types of phase transitions in the ferroelectric liquid crystal system.

Crystal structure of imidazole glycerol phosphate synthase: a tunnel through a (beta/alpha)8 barrel joins two active sites.

BACKGROUND: Imidazole glycerol phosphate synthase catalyzes a two-step reaction of histidine biosynthesis at the bifurcation point with the purine de novo pathway. The enzyme is a new example of intermediate channeling by glutamine amidotransferases in which ammonia generated by hydrolysis of glutamine is channeled to a second active site where it acts as a nucleophile. In this case, ammonia reacts in a cyclase domain to produce imidazole glycerol phosphate and an intermediate of purine biosynthesis. The enzyme is also a potential target for drug and herbicide development since the histidine pathway does not occur in mammals. RESULTS: The 2.1 A crystal structure of imidazole glycerol phosphate synthase from yeast reveals extensive interaction of the glutaminase and cyclase catalytic domains. At the domain interface, the glutaminase active site points into the bottom of the (beta/alpha)(8) barrel of the cyclase domain. An ammonia tunnel through the (beta/alpha)(8) barrel connects the glutaminase docking site at the bottom to the cyclase active site at the top. A conserved "gate" of four charged residues controls access to the tunnel. CONCLUSIONS: This is the first structure in which all the components of the ubiquitous (beta/alpha)(8) barrel fold, top, bottom, and interior, take part in enzymatic function. Intimate contacts between the barrel domain and the glutaminase active site appear to be poised for crosstalk between catalytic centers in response to substrate binding at the cyclase active site. The structure provides a number of potential sites for inhibitor development in the active sites and in a conserved interdomain cavity.

Human DNA replication initiation factors, ORC and MCM, associate with oriP of Epstein-Barr virus.

The 165-kb chromosome of Epstein-Barr virus (EBV) is replicated by cellular enzymes only once per cell cycle in human cells that are latently infected. Here, we report that the human origin recognition complex, ORC, can be detected in association with an EBV replication origin, oriP, in cells by using antibodies against three different subunits of human ORC to precipitate crosslinked chromatin. Mcm2, a subunit of the MCM replication licensing complex, was found to associate with oriP during G(1) and to dissociate from it during S phase. The detection of ORC and Mcm2 at oriP was shown to require the presence of the 120-bp replicator of oriP. Licensing and initiation of replication at oriP of EBV thus seem to be mediated by ORC. This is an example of a virus apparently using ORC and associated factors for the propagation of its genome.