Search

Reprogramming somatic fibroblasts into alternative lineages would provide a promising source of cells for regenerative therapy. However, transdifferentiating human cells into specific homogeneous, functional cell types is challenging. Here we show that cardiomyocyte-like cells can be generated by treating human fibroblasts with a combination of nine compounds that we term 9C. The chemically induced cardiomyocyte-like cells uniformly contracted and resembled human cardiomyocytes in their transcriptome, epigenetic, and electrophysiological properties. 9C treatment of human fibroblasts resulted in a more open-chromatin conformation at key heart developmental genes, enabling their promoters and enhancers to bind effectors of major cardiogenic signals. When transplanted into infarcted mouse hearts, 9C-treated fibroblasts were efficiently converted to chemically induced cardiomyocyte-like cells. This pharmacological approach to lineage-specific reprogramming may have many important therapeutic implications after further optimization to generate mature cardiac cells.

Field data on fault-slip observations is commonly heterogeneous. Paleostress estimation from such data sets is, in general, carried out in two steps: (i) the classification of the heterogeneous data set into homogeneous subsets and (ii) an inversion of each homogeneous subset. This study gives a new approach, the HGA, that combines the two issues in a single step process and gives the stress tensors directly. The given heterogeneous data are directly operated upon by the genetic algorithm operators, initialization, elitism, selection, encoding, crossover and mutation. These operations simulate such a guided search that finds successively fitter solutions, the stress tensors, until the globally fittest solution is obtained. We first explain the basic steps of the algorithm on a working example and then demonstrate its veracity using several synthetic and two natural examples. The proposed genetic algorithm method obviates the necessity of having first to classify the heterogeneous data into homogeneous sets. It directly estimates different stress states by inversion of the given heterogeneous fault-slip data. In contrast to the existing linear methods, the method is not vulnerable to entrapment of the solution in a local optimum. Although the method requires an a priori estimate of the maximum number of expected homogeneous sets in a given population, this estimate does not control the final results. Like any other method, the genetic algorithm method too has its merits and limitations and these are discussed.

We demonstrate the first planar Airy light-sheet microscope. Fluorescence light-sheet microscopy has become the method of choice to study large biological samples with cellular or sub-cellular resolution. The propagation-invariant Airy beam enables a ten-fold increase in field-of-view with single-photon excitation; however, the characteristic asymmetry of the light-sheet limits its potential for multi-photon excitation. Here we show how a planar light-sheet can be formed from the curved propagation-invariant Airy beam. The resulting symmetric light sheet excites two-photon fluorescence uniformly across an extended field-of-view without the need for deconvolution. We demonstrate the method for rapid two-photon imaging of large volumes of neuronal tissue., Comment: 7 pages, 4 figures

Apolipoprotein E (APOE) is a multifunctional protein that plays significant roles in important cellular mechanisms in peripheral tissues and is as well expressed in the central nervous system, notably by adult neural stem cells (NSCs) in the hippocampus. Evidence from animal studies suggest that APOE is critical for adult NSC maintenance. However, whether APOE has the potential to play a similar role in human NSCs has not been directly investigated. To address this question, we conducted a focused study characterising APOE gene and protein expression in an in vitro model of neural differentiation utilising human induced pluripotent stem cells. We found that APOE gene expression was dramatically decreased as the cells became more differentiated, indicating that APOE expression levels reflect the degree of cellular differentiation during neural induction. Furthermore, qualitative analysis results of immunocytochemistry showed that intracellular localisation of APOE protein becomes more pronounced as neural differentiation progresses. Taken together, our findings suggest a potential role for APOE in human NSC maintenance and justify further investigations being carried out to understand whether changes in APOE levels can directly impact the neurogenic capacity of human stem cells.

Organogenesis involves integration of myriad cell types with reciprocal interactions, each progressing through successive stages of lineage specification and differentiation. Establishment of unique gene networks within each cell dictates fate determination, and mutations of transcription factors that drive such networks can result in birth defects. Congenital heart defects are the most common malformations and are caused by disruption of discrete subsets of progenitors1–3, however, determining the transcriptional changes in individual cells that lead to organ-level defects in the heart, or other organs, has not been tractable. Here, we employed single-cell RNA sequencing to interrogate early cardiac progenitor cells as they become specified during normal and abnormal cardiogenesis, revealing how dysregulation of specific cellular sub-populations can have catastrophic consequences. A network-based computational method for single-cell RNA-sequencing that predicts lineage specifying transcription factors4,5identifiedHand2as a specifier of outflow tract cells but not right ventricular cells, despite failure of right ventricular formation inHand2-null mice6. Temporal single-cell transcriptome analysis ofHand2-null embryos revealed failure of outflow tract myocardium specification, whereas right ventricular myocardium differentiated but failed to migrate into the anterior pole of the developing heart. Dysregulation of retinoic acid signaling, responsible for anterior-posterior patterning7, was associated with posteriorization of anterior cardiac progenitors inHand2-null mutant hearts and ectopic atrial gene expression in outflow tract and right ventricle precursors. This work reveals transcriptional determinants in individual cells that specify cardiac progenitor cell fate and differentiation and exposes mechanisms of disrupted cardiac development at single-cell resolution, providing a framework to investigate congenital heart defects.

Although gene discovery in neuropsychiatric disorders, including autism spectrum disorder, intellectual disability, epilepsy, schizophrenia, and Tourette disorder, has accelerated, resulting in a large number of molecular clues, it has proven difficult to generate specific hypotheses without the corresponding datasets at the protein complex and functional pathway level. Here, we describe one path forward--an initiative aimed at mapping the physical and genetic interaction networks of these conditions, and then using these maps to connect the genomic data to neurobiology and, ultimately, the clinic. These efforts will include a team of geneticists, structural biologists, neurobiologists, systems biologists, and clinicians, leveraging a wide array of experimental approaches and creating a collaborative infrastructure necessary for long-term investigation. This initiative will ultimately intersect with parallel studies that focus on other diseases, as there is a significant overlap with genes implicated in cancer, infectious disease, and congenital heart defects.

Multiple tools have been developed to identify copy number variants (CNVs) from whole exome (WES) and whole genome sequencing (WGS) data. Current tools such as XHMM for WES and CNVnator for WGS identify CNVs based on changes in read depth. For WGS, other methods to identify CNVs include utilizing discordant read pairs and split reads and genome-wide local assembly with tools such as Lumpy and SvABA, respectively. Here, we introduce a new method to identify deletion CNVs from WES and WGS trio data based on the clustering of Mendelian errors (MEs). Using our Mendelian Error Method (MEM), we identified 127 deletions (inherited and de novo) in 2,601 WES trios from the Pediatric Cardiac Genomics Consortium, with a validation rate of 88% by digital droplet PCR. MEM identified additional de novo deletions compared to XHMM, and also identified sample switches, DNA contamination, a significant enrichment of 15q11.2 deletions compared to controls and eight cases of uniparental disomy. We applied MEM to WGS data from the Genome In A Bottle Ashkenazi trio and identified deletions with 97% specificity. MEM provides a robust, computationally inexpensive method for identifying deletions, and an orthogonal approach for verifying deletions called by other tools.

Estrogens have been shown to rapidly regulate local signalling events at both synapses and within the nucleus. The result of these signalling events is to rapidly modulate synapse structure and function, as well as epigenetic mechanisms including histone modifications. Ultimately these mechanisms are thought to contribute to long-lasting changes in neural circuitry, and thus influences cognitive functions such as learning and memory. However, the mechanisms by which estrogen-mediated local synaptic and nuclear signalling events are coordinated are not well understood. In this study we have found that the scaffold protein afadin, (also known as AF-6), undergoes a bi-directional trafficking to both synaptic and nuclear compartment in response to acute 17β-estradiol (estradiol) treatment. Interestingly, nuclear accumulation of afadin was coincidental with an increase in the phosphorylation of histone H3 at serine 10 (H3S10p). This histone modification is associated with the remodelling of chromatin into an open euchromatin state, allowing for transcriptional activation and related learning and memory processes. Critically, the cyto-nuclear trafficking of afadin was required for estradiol-dependent H3S10p. We further determined that nuclear accumulation of afadin is sufficient to induce phosphorylation of the mitogentic kinases ERK1/2 (pERK1/2) within the nucleus. Moreover, nuclear pERK1/2 was required for estradiol-dependent H3S10p. Taken together, we propose a model whereby estradiol incudes the bi-directional trafficking of afadin to synaptic and nuclear sub-compartments. Within the nucleus, afadin is required for increased pERK1/2 which in turn is required for H3S10p. Therefore this represents a mechanism through which estrogens may be able to coordinate both synaptic and nucleosomal events within the same neuronal population.

Introduction Synapse loss is the structural correlate of the cognitive decline indicative of dementia. In the brains of Alzheimer's disease sufferers, amyloid β (Aβ) peptides aggregate to form senile plaques but as soluble peptides are toxic to synapses. We previously demonstrated that Aβ induces Dickkopf-1 (Dkk1), which in turn activates the Wnt–planar cell polarity (Wnt-PCP) pathway to drive tau pathology and neuronal death. Methods We compared the effects of Aβ and of Dkk1 on synapse morphology and memory impairment while inhibiting or silencing key elements of the Wnt-PCP pathway. Results We demonstrate that Aβ synaptotoxicity is also Dkk1 and Wnt-PCP dependent, mediated by the arm of Wnt-PCP regulating actin cytoskeletal dynamics via Daam1, RhoA and ROCK, and can be blocked by the drug fasudil. Discussion Our data add to the importance of aberrant Wnt signaling in Alzheimer's disease neuropathology and indicate that fasudil could be repurposed as a treatment for the disease., Highlights • Aβ synaptotoxicity is Dickkopf-1 and Wnt-PCP dependent. • The Wnt-PCP pathway drives Aβ-driven synapse loss via RhoA and ROCK. • ROCK inhibitor fasudil blocks Aβ-driven synapse loss and cognitive impairment. • Fasudil should be assessed for repurposing for Alzheimer's disease.

Background The human neural progenitor cell line CTX0E16 is a robust source of forebrain-like glutamatergic cortical neurons that display intrinsic functional properties (Anderson et al., 2015, Stem Cell Res). This cell line was obtained from the embryonic cortex of a 12-week gestation XY fetus, and conditionally immortalised by ectopic expression of the c-MycER TAM transgene. This construct allows the stem cells to be maintained in a proliferative state under the presence of hydroxytamoxifen (4-OHT), while removal of 4-OHT and growth factors stimulates progenitor differentiation. The cortex is generally smaller in schizophrenia patients, and understanding the global gene expression changes occurring during cortical development, and its relationship to genetic risk for schizophrenia, might reveal clues as to which genetic mechanisms are responsible for mediating the differences observed in patients. Our hypothesis is that neurodevelopmental diseases such as schizophrenia might share a genetic component with processes driving early cortical maturation, which can be modelled in vitro. Methods The CTX0E16 cell line was provided by ReNeuron and total RNA was extracted from proliferating neural progenitor cells (NPCs, n=3) and from cells that underwent a 28-day differentiation protocol (n=3). RNA samples were then processed on Illumina HT12 v4 chips. Raw fluorescence probe intensity values were extracted using GenomeStudio, and were subsequently background-corrected and log-transformed in R using the lumi package. Differentially expressed probes were determined using t-tests and the false-discovery rate of multiple testing correction was applied (q Results We expect to identify and annotate sets of genes involved in cortical maturation that are additionally involved in the aetiology of schizophrenia. Discussion The discussion will focus on the role of cortical maturation as a process that may be involved in schizophrenia aetiology.

The fundamental role of the brain-specific myelin transcription factor 1-like (MYT1L) gene in cases of intellectual disability and in the aetiology of neurodevelopmental disorders is increasingly recognised. Yet, its function remains under-investigated. Here, we identify a network of helix-loop-helix (HLH) transcriptional regulators controlled by MYT1L, as indicated by our analyses in human neural stem cells and in the human brain. Using cell-based knockdown approaches and microarray analyses we found that (i) MYT1L is required for neuronal differentiation and identified ID1, a HLH inhibitor of premature neurogenesis, as a target. (2) While MYT1L prevented expression of ID1, it induced expression of a large number of terminal differentiation genes. (3) Consistently, expression of MYT1L in the human brain coincided with neuronal maturation and inversely correlated with that of ID1 and ID3 throughout the lifespan. (4) Genetic polymorphisms that reduced expression of MYT1L in the hippocampus resulted in increased expression of ID1 and ID3, decreased levels of the proneural basic HLH (bHLH) transcriptional regulators TCF4 and NEUROD6 and decreased expression of genes involved in long-term potentiation and synaptic transmission, cancer and neurodegeneration. Furthermore, our neuroimaging analyses indicated that MYT1L expression associated with hippocampal volume and activation during episodic memory recall, as measured by blood-oxygen-level dependent (BOLD) signals. Overall, our findings suggest that MYT1L influences memory-related processes by controlling a neuronal proliferation/differentiation switch of ID-bHLH factors.Neuropsychopharmacology accepted article preview online, 04 May 2017. doi:10.1038/npp.2017.91.

Human adult cardiomyocytes have limited regenerative capacity resulting in permanent loss of cardiomyocytes in the setting of many forms of heart disease. In an effort to replace lost cells, several groups have reported successful reprogramming of fibroblasts into induced cardiomyocyte-like cells (iCMs) without going through an intermediate progenitor or stem cell stage in murine and human models. This direct cardiac reprogramming approach holds promise as a potential method for regenerative medicine in the future and for dissecting the regulatory control of cell fate determination. Here we review the recent advances in the direct cardiac reprogramming field and the challenges that must be overcome to move this strategy closer to clinical application.

Single cardiomyocytes contain myofibrils that harbor the sarcomere-based contractile machinery of the myocardium. Cardiomyocytes differentiated from human pluripotent stem cells (hPSC-CMs) have potential as an in vitro model of heart activity. However, their fetal-like misalignment of myofibrils limits their usefulness for modeling contractile activity. We analyzed the effects of cell shape and substrate stiffness on the shortening and movement of labeled sarcomeres and the translation of sarcomere activity to mechanical output (contractility) in live engineered hPSC-CMs. Single hPSC-CMs were cultured on polyacrylamide substrates of physiological stiffness (10 kPa), and Matrigel micropatterns were used to generate physiological shapes (2,000-µm(2) rectangles with length:width aspect ratios of 5:1-7:1) and a mature alignment of myofibrils. Translation of sarcomere shortening to mechanical output was highest in 7:1 hPSC-CMs. Increased substrate stiffness and applied overstretch induced myofibril defects in 7:1 hPSC-CMs and decreased mechanical output. Inhibitors of nonmuscle myosin activity repressed the assembly of myofibrils, showing that subcellular tension drives the improved contractile activity in these engineered hPSC-CMs. Other factors associated with improved contractility were axially directed calcium flow, systematic mitochondrial distribution, more mature electrophysiology, and evidence of transverse-tubule formation. These findings support the potential of these engineered hPSC-CMs as powerful models for studying myocardial contractility at the cellular level.

© 2015 Elsevier Inc. The mechanisms by which transcription factor haploinsufficiency alters the epigenetic and transcriptional landscape in human cells to cause disease are unknown. Here, we utilized human induced pluripotent stem cell (iPSC)-derived endothelial cells (ECs) to show that heterozygous nonsense mutations in NOTCH1 that cause aortic valve calcification disrupt the epigenetic architecture, resulting in derepression of latent pro-osteogenic and -inflammatory gene networks. Hemodynamic shear stress, which protects valves from calcification in vivo, activated anti-osteogenic and anti-inflammatory networks in NOTCH1+/+, but not NOTCH1+/-, iPSC-derived ECs. NOTCH1 haploinsufficiency altered H3K27ac at NOTCH1-bound enhancers, dysregulating downstream transcription of more than 1,000 genes involved in osteogenesis, inflammation, and oxidative stress. Computational predictions of the disrupted NOTCH1-dependent gene network revealed regulatory nodes that, when modulated, restored the network toward the NOTCH1+/+state. Our results highlight how alterations in transcription factor dosage affect gene networks leading to human disease and reveal nodes for potential therapeutic intervention.

The present research work focuses on the mechanical, chemical, and curing characteristics of novolac resin based on renewable resource materials such as cardanol and furfural. Cardanol, a metasubstituted phenol, is a renewable organic resource obtained as a byproduct of the cashew industry. Furfural, an aromatic aldehyde, is also a renewable resource obtained as an agricultural waste product. Novolac resin has been synthesized by the condensation of cardanol with furfural in the presence of an oxalic acid catalyst and using varied molar proportions of the reacting monomers. The reaction was performed at 120°C. The progress of the reaction was monitored by determining the free formaldehyde and free phenol content. The prepared cardanol–furfural-based novolac resins were further characterized by various techniques such as infrared and nuclear magnetic resonance spectroscopic analysis. The resins were cured using the most suitable agent, hexamethylenetetramine. The differential scanning calorimetric technique was used to investigate the curing behavior of the prepared samples. The cured film samples were used for the determination of mechanical properties such as adhesion, flexibility, scratch hardness, gloss, and impact resistance; these cured film samples were also used to observe the effect of various chemicals and solvents (chemical resistance properties) such as sulfuric acid, acetic acid, sodium hydroxide, sodium carbonate and methanol, methyl ethyl ketone, xylene, and deionized water, respectively, on the surface of the film. The resulting coatings based on prepared cardanol–furfural resin were found to have excellent mechanical and chemical resistance properties.

Cardanol-based vinyl ester resin ( CVER ) was blended with varying concentrations of Carboxyl terminated butadiene acrylonitrile (CTBN) ranging between 0 and 15 wt% with an interval increment of 5 wt%, by two different approaches, viz., physical and chemical blending. The formation of various products during the curing of blend samples has been studied by FTIR spectroscopic analysis. The dynamic DSC scans showed that the blend samples of CVERs cured in the temperature range of 100–150 °C. It was observed that the blend sample containing 5 wt% and 10 wt% liquid rubber, for physical and chemical blending, respectively, showed least cure time, at 120 °C, amongst all other blend samples. On evaluation, it was found that in toughened samples thermal and mechanical properties were marginally affected with the increase of CTBN concentration. The blend morphology, studied by scanning electron microscopy analysis, showed the presence of precipitated discrete rubber particles which dispersed throughout the vinyl ester matrix, i.e., they revealed the presence of two phase morphology.

Dendritic spines were first described by Santiago Ramon y Cajal more than one hundred years ago when he examined Golgi-stained cerebellar Purkinje cells of birds. Since then, considerable effort has been put towards understanding how these structures are formed and what their functions in the central nervous system are. It is now well established that dendritic spines represent specialized subcellular compartments on the postsynaptic neuron where the majority of excitatory synapses are located. Therefore, the density of dendritic spines is a rough indication of how much excitatory input a particular neuron receives. One notable feature of these structures is the large heterogeneity of their dimensions and shapes. They can exist as short stubby spines, long thin spines, and mushroom-shaped spines. Moreover, they are highly dynamic, such that ongoing spine growth, turnover, and morphological changes occur in both developing and adult brains. Although excitatory synapses can form and function outside of dendritic spines, their location on spine heads likely confers additional properties. For example, the presence of the spine neck is thought to create an isolated biochemical compartment on the spine head, where individual synapses of the postsynaptic neuron can function and be regulated independently of each other. Changes in the dimension and shape of individual spines also allow modulation of synaptic efficacy between specific neuronal partners and therefore contribute to synaptic plasticity and provide the cellular basis of learning and memory. Indeed, many molecular players that regulate dendritic spine morphogenesis also turn out to be essential for learning-related synaptic plasticity and memory formation. In this special issue, reviews and original research papers have been collected to address various questions on dendritic spine biology. These include the process of spine development, the functional differentiation of large and small spines, the relationships between spine changes and learning, the signaling pathways that control spine morphogenesis, and the link between spine abnormalities and brain disorders.

The heart is a complex organ, consisting of multiple cell types that coordinately regulate blood flow. Reciprocal Notch pathway signalling in endocardial and myocardial cells is now shown to promote maturation of the ventricular chambers. These insights reveal mechanisms that, when disrupted, can lead to cardiomyopathies.

In the present article, the brief biography of Sir George Gabriel Stokes and his contribution to the field of physics and fluid mechanics has been reviewed.

Molecular chaperones regulate quality control in the human proteome, pathways that have been implicated in many diseases, including heart failure. Mutations in the BAG3 gene, which encodes a co-chaperone protein, have been associated with heart failure due to both inherited and sporadic dilated cardiomyopathy. Familial BAG3 mutations are autosomal dominant and frequently cause truncation of the coding sequence, suggesting a heterozygous loss-of-function mechanism. However, heterozygous knockout of the murine BAG3 gene did not cause a detectable phenotype. To model BAG3 cardiomyopathy in a human system, we generated an isogenic series of human induced pluripotent stem cells (iPSCs) with loss-of-function mutations in BAG3. Heterozygous BAG3 mutations reduced protein expression, disrupted myofibril structure, and compromised contractile function in iPSC-derived cardiomyocytes (iPS-CMs). BAG3-deficient iPS-CMs were particularly sensitive to further myofibril disruption and contractile dysfunction upon exposure to proteasome inhibitors known to cause cardiotoxicity. We performed affinity tagging of the endogenous BAG3 protein and mass spectrometry proteomics to further define the cardioprotective chaperone complex that BAG3 coordinates in the human heart. Our results establish a model for evaluating protein quality control pathways in human cardiomyocytes and their potential as therapeutic targets and susceptibility factors for cardiac drug toxicity.

Despite current standard of care, the average 5-year mortality after an initial diagnosis of heart failure (HF) is about 40%, reflecting an urgent need for new therapeutic approaches. Previous studies demonstrated that the epigenetic reader protein bromodomain-containing protein 4 (BRD4), an emerging therapeutic target in cancer, functions as a critical coactivator of pathologic gene transactivation during cardiomyocyte hypertrophy. However, the therapeutic relevance of these findings to human disease remained unknown. We demonstrate that treatment with the BET bromodomain inhibitor JQ1 has therapeutic effects during severe, preestablished HF from prolonged pressure overload, as well as after a massive anterior myocardial infarction in mice. Furthermore, JQ1 potently blocks agonist-induced hypertrophy in human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). Integrated transcriptomic analyses across animal models and human iPSC-CMs reveal that BET inhibition preferentially blocks transactivation of a common pathologic gene regulatory program that is robustly enriched for NFκB and TGF-β signaling networks, typified by innate inflammatory and profibrotic myocardial genes. As predicted by these specific transcriptional mechanisms, we found that JQ1 does not suppress physiological cardiac hypertrophy in a mouse swimming model. These findings establish that pharmacologically targeting innate inflammatory and profibrotic myocardial signaling networks at the level of chromatin is effective in animal models and human cardiomyocytes, providing the critical rationale for further development of BET inhibitors and other epigenomic medicines for HF.

Diseases caused by gene haploinsufficiency in humans commonly lack a phenotype in mice that are heterozygous for the orthologous factor, impeding the study of complex phenotypes and critically limiting the discovery of therapeutics. Laboratory mice have longer telomeres relative to humans, potentially protecting against age-related disease caused by haploinsufficiency. Here, we demonstrate that telomere shortening in NOTCH1-haploinsufficient mice is sufficient to elicit age-dependent cardiovascular disease involving premature calcification of the aortic valve, a phenotype that closely mimics human disease caused by NOTCH1 haploinsufficiency. Furthermore, progressive telomere shortening correlated with severity of disease, causing cardiac valve and septal disease in the neonate that was similar to the range of valve disease observed within human families. Genes that were dysregulated due to NOTCH1 haploinsufficiency in mice with shortened telomeres were concordant with proosteoblast and proinflammatory gene network alterations in human NOTCH1 heterozygous endothelial cells. These dysregulated genes were enriched for telomere-contacting promoters, suggesting a potential mechanism for telomere-dependent regulation of homeostatic gene expression. These findings reveal a critical role for telomere length in a mouse model of age-dependent human disease and provide an in vivo model in which to test therapeutic candidates targeting the progression of aortic valve disease.

Background: Reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells in situ represents a promising strategy for cardiac regeneration. A combination of 3 cardiac transcription factors, Gata4, Mef2c, and Tbx5 (GMT), can convert fibroblasts into induced cardiomyocyte-like cells, albeit with low efficiency in vitro. Methods: We screened 5500 compounds in primary cardiac fibroblasts to identify the pathways that can be modulated to enhance cardiomyocyte reprogramming. Results: We found that a combination of the transforming growth factor-β inhibitor SB431542 and the WNT inhibitor XAV939 increased reprogramming efficiency 8-fold when added to GMT-overexpressing cardiac fibroblasts. The small molecules also enhanced the speed and quality of cell conversion; we observed beating cells as early as 1 week after reprogramming compared with 6 to 8 weeks with GMT alone. In vivo, mice exposed to GMT, SB431542, and XAV939 for 2 weeks after myocardial infarction showed significantly improved reprogramming and cardiac function compared with those exposed to only GMT. Human cardiac reprogramming was similarly enhanced on transforming growth factor-β and WNT inhibition and was achieved most efficiently with GMT plus myocardin. Conclusions: Transforming growth factor-β and WNT inhibitors jointly enhance GMT-induced direct cardiac reprogramming from cardiac fibroblasts in vitro and in vivo and provide a more robust platform for cardiac regeneration.

This paper presents the electron beam misalignment study with respect to cathode position and cathode magnetic field of 42 GHz, 200 kW gyrotron. The performance of gyrotron is affected with the misalignment of cathode position. The simulation results confirm the tolerance of cathode misalignment with respect to the design parameters such as the transverse-to-axial velocity ratio, the maximum transverse velocity spread, etc.

In a recent paper, Srivastava et al. (Int J Appl Math Mech 8:17–53, 2012) tackled the problem of steady Stokes flow around deformed sphere wherein author evaluated the general expression of drag with the help of method developed by Datta and Srivastava (Proc Indian Acad Sci Math Sci 109:441–452, 1999) in both situations when axially symmetric isolated body is placed under axial flow (along the axis of symmetry) and transverse flow(along perpendicular to axis of symmetry) corrected up to the second order of deformation parameter and applied those on to class of oblate bodies and to flat circular disk as a special case. In this paper, rotation of deformed sphere has been dealt in axial or longitudinal situation in which uniform stream is along the axis of symmetry under the no-slip boundary conditions. The general expression of torque in terms of axial Stokes drag (Srivastava et al., Int J Appl Math Mech 8:17–53, 2012) is produced with the help of method developed by Datta and Srivastava (Proc Indian Acad Sci Math Sci 109:441–452, 1999). The general form of deformed sphere, governed by polar equation, $$r = a(1 + \varepsilon \sum\nolimits_{k = 0}^{\infty } {d_{k} P_{k} ({ \cos } \theta )} )$$ (where, d k is shape factor and P k is Legendre function of first kind), is considered for the study. The expression of torque for slowly rotating deformed sphere is derived up to the order of O(e 2). The class of oblate and prolate axisymmetric bodies is considered for the validation and further numerical discussion. All the expressions of moment coefficients are updated up to the order of O(e 2), where e is deformation parameter. In particular, up to the order of O(e) and O(e 2), the numerical values of moment coefficients and their ratio have been evaluated for the various values of deformation parameters(e) and aspect ratio(b/a) for a class of oblate and prolate axially symmetric bodies including disk and thin slender elongated bodies as a special cases.

Cardanol-based vinyl ester resin (CVER) was prepared by reacting indigenously synthesized cardanol-based epoxidized novolac resin (CENR) with methacrylic acid (MA) in the presence of triphenylphosphine as catalyst. Five samples of cardanol-based vinyl ester resin containing styrene and glycidyl methacrylate (GMA), as diluents, in the weight ratios 40:0, 30:10, 20:20, 10:30 and 0:40 were prepared at room temperature. Sharp exotherms were observed in DSC scans in the temperature range of 60–170 °C. The onset temperature ( T onset ), peak exothermic temperature ( T p ) and completion temperature ( T stop ) decreased with increase in GMA content in the ternary blend systems of CVER/styrene/GMA. A broad exotherm was observed after the initial sharp exotherm that was attributed to the etherification reaction. Cured samples were found to be stable up to 205–235 °C and started loosing weight above this temperature. Rapid decomposition was observed in the temperature range of 400–550 °C as evidenced by TGA analysis. Increase of GMA content in ternary blend systems of CVER/styrene/GMA lowered the tensile strength progressively and enhanced the impact strength and elongation-at-break. The cured films of VER containing mixture of styrene and GMA exhibited good gloss and impact resistance. The chemical resistance of cured films of VER containing mixture of styrene and GMA showed good resistance to acids, deionized water, synthetic sea water and mineral turpentine oil.

The present paper reports the preparation and thermo-mechanical characterization of epoxy resin from renewable resource materials viz. cardanol and furfural. The cardanol furfural based epoxy resin was prepared by reacting cardanol furfural based novolac resin with molar excess of epichlorohydrin, in basic medium, at 120 °C. The cardanol furfural based novolac resin was synthesized by reacting cardanol and furfural in the molar ratios viz. 1.0:0.5, 1.0:0.6, 1.0:0.7 and 1.0:0.8, respectively, using succinic acid as catalyst. The prepared resin samples were characterized by Fourier-transform infrared and nuclear magnetic resonance spectroscopic analysis. The prepared epoxy resin samples were cured by using diaminodiphenylmethane. The samples were cured at 160 °C, in an air oven, which was determined by peak temperatures from the dynamic differential scanning calorimetric (DSC) scans. DSC technique was used to investigate the curing behavior. Thermal stability, as assesed by dynamic thermogravimetric traces of the epoxy resin sample with molar ratio 1:0.8, was found to be the highest among all other samples. The castings of the cured samples were used for the determination of mechanical properties such as tensile strength, impact strength etc. whereas cured film samples were used to observe the effect of various chemicals and solvents on the surface of the film. The epoxy resin sample prepared from high molar ratio i.e. 1:0.8 was found to exhibit better mechanical and chemical properties than other samples.

Resoles were prepared under microwave irradiation with different phenols, such as phenol, o-, p-, and m-cresols, separately with formaldehyde having formaldehyde/phenol ratio of 2:1 in basic medium. Analogical synthesis was performed using conventional heating for comparing the methods. The methylolation of phenol was confirmed by Fourier transform infrared spectroscopic analysis and a reaction mechanism was proposed. The number-average molecular weight was found by gel permeation chromatography technique. On the basis of the calculated value of kinetic chain length, the structure of the resole-type phenolic resin was proposed. Differential scanning calorimetry technique was used to investigate the curing behavior. As assessed by dynamic thermogravimetry, traces of resole sample prepared from p-cresol were found to possess better thermal stability, both in conventional as well as microwave-irradiated systems, among all other resole samples. The tensile strength, elongation at break, and impact strength showed an increasing trend. The main advantage of the process is about sixfold reduction of reaction time of the process carried at microwave reactors in comparison with the conventional heating.

The hydrology of high altitude glacierized basin in Himalaya is complex to understand due to high variability in climate and lack of hydro-meteorological data. In this study, analysis of hydrological, glacier ablation and meteorological records have been made for Dunagiri Glacier (~4,200 m) during the melt season (July - September), for the years 1985 and 1987–1989. In the daily cycle of mean diurnal discharge in summer months during 1985–1989, maximum discharge occurred at midnight, and minimum discharge occurred in the morning. Mean daytime and nighttime discharge volume was contributing approximately equal amounts. Such type of analysis will be the basis for designing/developing Run-off-the river type hydroelectric power projects which are heavily dependent on snow and glacier melt in Himalaya. Time series analysis of available hydro-meteorological records have been used to understand governing hydrological processes within the basin and to develop regression model for future development in runoff forecasting using climate data.

Fry method is a graphical technique that displays the strain ellipse in the form of central vacancy on a point distribution, the Fry plot. For objective strain estimation from the Fry plot, the central vacancy must appear as a sharply focused ellipse. In practice, however, a diffused appearance of the central vacancy in the Fry plots induces considerable subjectivity in direct strain estimation. Several alternative computer-based methods have recently been proposed for objective strain estimation from the Fry plots. Relative merits and limitations of these methods are, however, not yet well understood. This study compares the accuracy and efficacy of the six recent methods proposed for objective strain estimation from the Fry plot. The six methods are based on four different approaches, the point density contrast, the image analysis, the Delaunay triangulation and the point distribution analysis. We test and compare these methods by: (i) generating variously sorted object sets through computer simulations, (ii) distorting the object sets, with the help of computer graphics software, by pure shear, simple shear and simultaneous pure-and-simple shear and, (iii) estimating the errors. Results from more than 2900 tests imply that the continuous function method and the Delaunay triangulation method yield most accurate strain estimates. The image analysis method gives very accurate principal strain orientations but overestimates the strain ratio, specially, in strongly distorted object sets. The point distribution analysis methods are successful mostly on those point distributions that are obtained by pure shear distortion of well-sorted objects with specific boundary conditions. With respect to the time-efficiency, the image analysis method and the Delaunay triangulation method are found to be most rapid.

An epoxy resin based on cardanol and varying content of CTBN was cured using polyamine as a hardener. The ultimate aim of the study was to modify the brittle epoxy matrix by the liquid rubber to improve toughness characteristics. FTIR of the modified was performed to understand the structural transformations taking place during the uncured and cured stage of the modified systems. Also, the structures were proposed on the basis of the results of NMR and MALDI-TOF mass spectroscopic analysis along with GPC analysis. Mechanical properties of neat as well as modified networks have been studied to observe the effect of rubber modification. The improvement in these properties indicates that the rubber-modified resin would be more durable than the pure epoxy based on cardanol. A clear-cut two-step mass loss in dynamic thermogravimetric trace of unmodified and CTBN-modified systems was observed. Modified cardanol based epoxy network displayed two phase separated morphology.

The esterification reaction of phenol-cardanol based epoxidized novolac-vinyl ester resins was performed in the temperature range of 80–120 °C with an interval of 10 °C using a 1:0.9, epoxy-to-carboxylic group ratio in the presence of triphenyl phosphine as catalyst. The results based on kinetic study, indicated that the reaction followed first order kinetics. FT-IR analysis of vinyl ester resins revealed that addition esterification reaction between carbonyl and epoxy group took place and no side chain reactions occurred during the synthesis. A comparison of mechanism based rate equation and experimental data showed an excellent agreement. Finally, Arrhenius equation and activation energy were presented. The specific rate constants, based on linear regression analysis, were found to obey Arrhenius equation. The values of activation energy, frequency factor, enthalpy, entropy, and free energy of the reaction revealed that the reaction was spontaneous and irreversible and produced a highly activated complex.

In this paper, Oseen?s correction to Stokes drag experienced by axially symmetric particle placed in a uniform stream perpendicular to axis of symmetry(i.e. transverse flow) is obtained. For this, the linear relationship between axial and transverse Stokes drag is utilized to extend the Brenner?s formula for axial flow to transverse flow. General expression of Oseen?s correction to Stokes drag on axially symmetric particle placed in transverse flow is found to be new. This general expression is applied to some known axially symmetric bodies and obtained values of Oseen?s drag, up to first order terms in Reynolds number ?R?, are also claimed to be new and never exist in the literature. Numerical values of Oseen drag are also evaluated and their variations with respect to Reynolds number, eccentricity and deformation parameter are depicted in figures and compared with some known values. Some important applications are also highlighted.

The method and the advantages of an evolutionary computing based approach using a steady state genetic algorithm (GA) for the parameterization of interatomic potentials for metal oxides within the shell model framework are developed and described. We show that the GA based methodology for the parameterization of interatomic force field functions is capable of (a) simultaneous optimization of the multiple phases or properties of a material in a single run, (b) facilitates the incremental re-optimization of the whole system as more data is made available for either additional phases or material properties not included in previous runs, and (c) successful global optimization in the presence of multiple local minima in the parameter space. As an example, we apply the method towards simultaneous optimization of four distinct crystalline phases of Barium Titanate (BaTiO3 or BTO) using an ab initio density functional theory (DFT) based reference dataset. We find that the optimized force field function is capable of the prediction of the two phases not used in the optimization procedure, and that many derived physical properties such as the equilibrium lattice constants, unit cell volume, elastic properties, coefficient of thermal expansion, and average electronic polarization are in good agreement with the experimental results available from the literature.

Cashew nut shell liquid (CNSL), an agricultural renewable resource material, produces natural phenolic distillates such as cardanol. Cardanol condenses with formaldehyde at the ortho- and para-position of the phenolic ring under acidic or alkaline condition to yield a series of polymers of novolac- or resol-type phenolic resins. These phenolic resins may further be modified by epoxidation with epichlorohydrin to duplicate the performance of such phenolic-type novolacs (CFN). The structural changes during curing of blend samples of epoxy and carboxyl terminated poly (butadiene-co-acrylonitrile) (CTBN) were studies by Fourier-transform infrared (FTIR) spectrophotometer. The epoxy samples were synthesized by biomass material, cardanol. Blend sample was prepared by physical mixing of CTBN ranging between 0 and 20 weight percent CTBN liquid rubber into cardanol-based epoxidized novolac (CEN) resin. The FTIR spectrum of uncured blend sample clearly indicated that there appeared a band in the region of 3200–3500 cm− 1 which might be due to the presence of phenolic hydroxyl group and OH group of the opened epoxide. Pure epoxy resin showed peaks near 856 cm− 1 which might be due to oxirane functionality of the epoxidized novolac resin. Both epoxy and its blend sample was cured with polyamine. The cure temperature of CEN resin was found to be decreased by the incorporation of CTBN. The decomposition behavior was also studied by thermogravimetric analyzer (TGA). Two-step decomposition behavior was observed in both epoxy and its blend samples.

Mutation of highly conserved residues in transcription factors may affect protein-protein or protein-DNA interactions, leading to gene network dysregulation and human disease. Human mutations in GATA4, a cardiogenic transcription factor, cause cardiac septal defects and cardiomyopathy. Here, iPS-derived cardiomyocytes from subjects with a heterozygous GATA4-G296S missense mutation showed impaired contractility, calcium handling, and metabolic activity. In human cardiomyocytes, GATA4 broadly co-occupied cardiac enhancers with TBX5, another transcription factor that causes septal defects when mutated. The GATA4-G296S mutation disrupted TBX5 recruitment, particularly to cardiac super-enhancers, concomitant with dysregulation of genes related to the phenotypic abnormalities, including cardiac septation. Conversely, theGATA4-G296S mutation led to failure of GATA4 and TBX5-mediated repression at non-cardiac genes and enhanced open chromatin states at endothelial/endocardial promoters. These results reveal how disease-causing missense mutations can disrupt transcriptional cooperativity, leading to aberrant chromatin states and cellular dysfunction, including those related to morphogenetic defects.

A combination of two groups of boudins, exposed in orthogonal sections, is observed as chocolate tablet structures in the South Almora Thrust (SAT) zone, Kumaun Lesser Himalaya. Evidence from boudin associated veins, fractures, folds and microstructures suggest that the chocolate-tablet boudins were developed as a consequence of layer-parallel stretching in two orthogonal directions during two successive phases of ductile shearing in the SAT zone. The early phase of top-to-NW shearing is speculated as a pre-Himalayan deformation during which the early boudins were developed. This deformation was superposed by a late phase of top-to-SW Himalayan shearing and development of the attendant late boudins. It is concluded that the interference of two phases of successively developed boudins has resulted into development of the chocolate tablet structures in the SAT. Fluid inclusion and microstructures indicate a low temperature, about 300 °C, growth of the chocolate-tablet boudins. The finite strain analysis reveals that a maximum strain ratio (Rs) of 1.7 was accumulated during the pre-Himalayan phase of deformation whereas Rs was of the order of 2.1 in the Himalayan phase of deformation.