Growth, self-assembly and dynamics of nano-scale films at fluid interfaces

Ultrathin films at fluid interfaces are important not only from a fundamental point of view as 2D complex fluids but have also become increasingly relevant in the development of novel functional materials. There has been an explosion in the synthesis work in this area over the last decade, giving rise to many exotic nanostructures at fluid interfaces. However, the factors controlling particle nucleation, growth and self-assembly at interfaces are poorly understood on a quantitative level. We will outline some of the recent attempts in this direction. Some of the selected investigations examining the macroscopic mechanical properties of molecular and particulate films at fluid interfaces will be reviewed. We conclude with a discussion of the electronic properties of these films that have potential technological and biological applications.

Changes in the Levels of Free Amino Acids in Various Regions of Cuscuta during Growth

The angiospermous plant parasite Cuscuta derives reduced carbon and nitrogen compounds primarily from its host. Free amino acids along Cuscuta vines in three zones, viz., 0 to 5 cm, 5 to 15 cm, and 15 to 30 cm, which in a broad sense represent the region of cell division, cell elongation and differentiation and vascular tissue differentiation respectively, were quantitatively estimated. The free amino acid content was the highest in the 0 to 5 cm region and progressively decreased along the posterior regions of the vine. The haustorial region showed the lowest content of free amino acids. In general, the free amino acid content in samples collected at 7 p.m. was found to be higher than that in the samples collected at 7 a.m. Three basic amino acids, histidine, the uncommon amino acid γ-hydroxyarginine, and arginine constituted more than 50% of the total free amino acids in all the zones studied except the haustorial region. Aspartic acid and glutamic acid constituted the major portion in the acidic and neutral fraction of amino acids. Glutamine, asparagine, threonine, and serine were eluted together and occurred in substantial amounts. γ-Hydroxyarginine constituted the largest fraction in the cut end exudate of Cuscuta and presumably appeared to be the major form of transport amino acid. γ-Hydroxyarginine was also a major constituent of the basic amino acids in Cuscuta vines parasitizing host plants from widely separated families, suggesting that this amino acid is a biosynthetic product of the parasite rather than that of the hosts. Also, U-14C arginine was converted to γ-hydroxyarginine by cut Cuscuta vines, suggesting that γ-hydroxyarginine is synthesized de novo from arginine by Cuscuta.

High-yield bacterial expression and structural characterization of recombinant human insulin-like growth factor binding protein-2

The diverse biological activities of the insulin-like growth factors (IGF-1 and IGF-2) are mediated by the IGF-1 receptor (IGF-1R). These actions are modulated by a family of six IGF-binding proteins (ICFBP-1-6; 22-31 kDa) that via high affinity binding to the IGFs (K-D similar to 300-700 pM) both protect the IGFs in the circulation and attenuate IGF action by blocking their receptor access In recent years, IGFBPs have been implicated in a variety of cancers However, the structural basis of their interaction with IGFs and/or other proteins is not completely understood A critical challenge in the structural characterization of full-length IGFBPs has been the difficulty in expressing these proteins at levels suitable for NMR/X-ray crystallography analysis Here we describe the high-yield expression of full-length recombinant human IGFBP-2 (rhIGFBP-2) in Eschericha coli Using a single step purification protocol, rhIGFBP-2 was obtained with >95% purity and structurally characterized using NMR spectroscopy. The protein was found to exist as a monomer at the high concentrations required for structural studies and to exist in a single conformation exhibiting a unique intra-molecular disulfide-bonding pattern The protein retained full biologic activity. This study represents the first high-yield expression of wild-type recombinant human IGFBP-2 in E coli and first structural characterization of a full-length IGFBP (C) 2010 Elsevier Inc. All rights reserved

Rapid burst of H2O2 by plant growth regulators increases intracellular Ca2+ amounts and modulates CD4(+) T cell activation

The identification of small molecules that affect T cell activation is an important area of research. Three molecules that regulate plant growth and differentiation, but not their structurally similar analogs, were identified to enhance primary mouse CD4(+) T cell activation in conjunction with soluble anti-CD3 stimulation: Indoleacetic acid (natural plant auxin), 1-Napthaleneacetic acid (synthetic plant auxin) and 2,4-Dichlorophenoxyacetic acid (synthetic plant auxin and herbicide). These effects are distinct in comparison to Curcumin, the well known phenolic immunomodulator, which lowers T cell activation. An investigation into the mechanisms of action of the three plant growth regulators revealed a rapid induction of reactive oxygen species (ROS), mainly comprising H2O2 . In addition, these three molecules synergize with soluble anti-CD3 signaling to enhance intracellular Ca2+ concentrations Ca2+](i), leading to greater T cell activation, e.g. induction of CD25 and IL-2. Enhanced production of TNF alpha and IFN gamma by CD4+ T cells is also observed upon plant growth regulator treatment with soluble anti-CD3. Interestingly, maximal IL-2 production and CD4(+) T cell cycle progression are observed upon activation with soluble anti-CD3 and phorbol 12-myristate 13-acetate (PMA), a phorbol ester. Additionally, stimulation with PMA and Ionomcyin (a Ca2+ ionophore), which activates T cells by circumventing the TCR, and plant growth regulators also demonstrated the role of the strength of signal (SOS): T cell cycle progression is enhanced with gentle activation conditions but decreased with strong activation conditions. This study demonstrates the direct effects of three plant growth regulators on CD4(+) T cell activation and cycling. (C) 2010 Elsevier B.V. All rights reserved.

Growth Kinetics of ZnO Nanocrystals in the Presence of a Base: Effect of the Size of the Alkali Cation

Following an earlier study (J. Am. Chem Soc. 2007, 129, 4470) describing a very unusual growth kinetics of ZnO nanoparticles, we critically evaluate here the proposed mechanism involving a crucial role of the alkali base ion in controlling the growth of ZnO nanoparticles using other alkali bases, namely, LiOH and KOH. While confirming the earlier conclusion of the growth of ZnO nanoparticles being hindered by an effective passivating layer of cations present in the reaction mixture and thereby generalizing this phenomenon, present experimental data reveal an intriguing nonmonotonic dependence of the passivation efficacy on the ionic size of the alkali base ion. This unexpected behavior is rationalized on the basis of two opposing factors: (a) solvated cationic radii and (b) dissociation constant of the base.

Effects of temper level on the dependence of fatigue crack growth threshold and crack closure on the prior austenitic grain size

An attempt has been made to systematically investigate the effects of microstructural parameters, such as the prior austenite grain size (PAGS), in influencing the resistance to fatigue crack growth (FCG) in the near-threshold region under three different temper levels in a quenched and tempered high-strength steel. By austenitizing at various temperatures, the PAGS was varied from about 0.7 to 96 μm. The microstructures with these grain sizes were tempered at 200 °C, 400 °C, and 530 °C and tested for fatigue thresholds and crack closure. It has been found that, in general, three different trends in the dependence of both the total threshold stress intensity range, ΔK th , and the intrinsic threshold stress intensity range, ΔK eff, th , on the PAGS are observable. By considering in detail the factors such as cyclic stress-strain behavior, environmental effects on FCG, and embrittlement during tempering, the present observations could be rationalized. The strong dependence of ΔK th and ΔK eff, th on PAGS in microstructures tempered at 530 °C has been primarily attributed to cyclic softening and thereby the strong interaction of the crack tip deformation field with the grain boundary. On the other hand, a less strong dependence of ΔK th and ΔK eff, th on PAGS is suggested to be caused by the cyclic hardening behavior of lightly tempered microstructures occurring in 200 °C temper. In both microstructures, crack closure influenced near-threshold FCG (NTFCG) to a significant extent, and its magnitude was large at large grain sizes. Microstructures tempered at the intermediate temperatures failed to show a systematic variation of ΔKth and ΔKeff, th with PAGS. The mechanisms of intergranular fracture vary between grain sizes in this temper. A transition from “microstructure-sensitive” to “microstructure-insensitive” crack growth has been found to occur when the zone of cyclic deformation at the crack tip becomes more or less equal to PAGS. Detailed observations on fracture morphology and crack paths corroborate the grain size effects on fatigue thresholds and crack closure.

Role of elasto-plastic analysis under cyclic loading in fatigue crack growth studies

Linear Elastic Fracture Mechanics (LEFM) has been widely used in the past for fatigue crack growth studies, but this is acceptable only in situations which are within small scale yielding (SSY). In many practical structural components, conditions of SSY could be violated and one has to look for fracture criteria based on elasto-plastic analysis. Crack closure phenomenon, one of the most striking discoveries based on inelastic deformations during crack growth, has significant effect on fatigue crack growth rate. Numerical simulation of this phenomenon is computationally intensive and involved but has been successfully implemented. Stress intensity factors and strain energy release rates lose their meaning, J-integral (or its incremental) values are applicable only in specific situations, whereas alternate path independent integrals have been proposed in the literature for use with elasto-plastic fracture mechanics (EPFM) based criteria. This paper presents certain salient features of two independent finite element (numerical) studies of relevance to fatigue crack growth, where elasto-plastic analysis becomes significant. These problems can only be handled in the current day computational environment, and would have been only a dream just a few years ago.

Molecular organization of the gene encoding Xenopus laevis transforming growth factor-beta 5

Transforming growth factor-beta s (TGF-beta 5) are multifunctional polypeptides, known to influence proliferation and differentiation of many cell types. TGF-beta 5 cDNA was cloned from Xenopus laevis and this isoform is unique to the amphibians. Here, we report the isolation and characterization of the TGF-beta 5 genomic clones to determine the structure of TGF-beta 5 gene. The gene consists of seven exons and all intron-exon boundaries follow the GT-AG consensus. The organization of TGF-beta 5 gene was identical to that of the mammalian TGF-beta isoforms, with the exception of position of the first splice junction. We determined the size of TGF-beta 5 gene to be approximately 20 kb.

Influence of growth below and above T-c on the morphology and domain structure in flux-grown KTP crystals

KTP crystals have been grown below and above the ferroelectric transition temperature by flux method employing both spontaneous and top-seeded solution growth techniques. A slight morphological difference has been observed in these crystals when grown below and above the T-c. Ferroelectric domains are studied in these crystals by selective domain etching. It is seen that the ferroelectric domains in crystals grown spontaneously below T, show a complicated structure. A systematic investigation of the factors influencing domain structure has been carried out. Stress to some extent has been shown to affect the domain structure. Finally, a convenient way of converting the multidomain crystals into monodomain ones is described.

Isolation of transforming growth factor-beta 2 cDNA from a fish, Cyprinus carpio by RT-PCR

Transforming Growth Factors-beta (TGF-beta s) have been described in many vertebrate species of amphibians, aves and mammals. In this report we demonstrate the presence of TGF-beta 2 in pisces. TGF-beta 2 has been cloned from a fish, Cyrinus carpio, by RT-PCR using degenerate oligonucleotide primers. Sequence analysis of the amplified product and alignment of the deduced amino acid sequence with the human TGF-beta 2 amino acid sequence revealed 81% and 93% identity in the precursor and the mature regions, respectively. The northern blot analysis of fish heart RNA shows a major messenger RNA species of about 8.0 kb and two messages of very low abundance of about 5.0 kb and 4.0 kb. The identification of TGF-beta 2 isoform in Pisces and it's high degree of homology with the mammalian isoform suggests that among all TGF-beta isoforms, TGF-beta 2 is the most conserved during evolution. (C) 1997 Elsevier Science B.V.

Growth inhibitory, apoptotic and anti-inflammatory activities displayed by a novel modified triterpenoid, cyano enone of methyl boswellates

Triterpenoids are pentacyclic secondary metabolites present in many terrestrial plants. Natural triterpenoids have been reported to exhibit anti-inflammatory and anti-carcinogenic activities. Here, we show that modifications of ring A of boswellic acid (2 cyano, 3 enone) resulted in a highly active growth inhibitory, anti-inflammatory, pro-differentiative and anti-tumour triterpenoid compound called cyano enone of methyl boswellates (CEMB). This compound showed cytotoxic activity on a number of cancer cell lines with IC50 ranging from 0.2 to 0.6 mu M. CEMB inhibits DNA synthesis and induces apoptosis in A549 cell line at 0.25 mu M and 1 mu M concentrations, respectively. CEMB induces adipogenic differentiation in 3T3-L1 cells at a concentration of 0.1 mu M. Finally, administration of CEMB intra-tumourally significantly inhibited the growth of C6 glioma tumour xenograft in immuno-compromised mice. Collectively, these results suggest that CEMB is a very potent anti-tumour compound.

Gene Expression Profiling of Preovulatory Follicle in the Buffalo Cow: Effects of Increased IGF-I Concentration on Periovulatory Events

The preovulatory follicle in response to gonadotropin surge undergoes dramatic biochemical, and morphological changes orchestrated by expression changes in hundreds of genes. Employing well characterized bovine preovulatory follicle model, granulosa cells (GCs) and follicle wall were collected from the preovulatory follicle before, 1, 10 and 22 h post peak LH surge. Microarray analysis performed on GCs revealed that 450 and 111 genes were differentially expressed at 1 and 22 h post peak LH surge, respectively. For validation, qPCR and immunocytochemistry analyses were carried out for some of the differentially expressed genes. Expression analysis of many of these genes showed distinct expression patterns in GCs and the follicle wall. To study molecular functions and genetic networks, microarray data was analyzed using Ingenuity Pathway Analysis which revealed majority of the differentially expressed genes to cluster within processes like steroidogenesis, cell survival and cell differentiation. In the ovarian follicle, IGF-I is established to be an important regulator of the above mentioned molecular functions. Thus, further experiments were conducted to verify the effects of increased intrafollicular IGF-I levels on the expression of genes associated with the above mentioned processes. For this purpose, buffalo cows were administered with exogenous bGH to transiently increase circulating and intrafollicular concentrations of IGF-I. The results indicated that increased intrafollicular concentrations of IGF-I caused changes in expression of genes associated with steroidogenesis (StAR, SRF) and apoptosis (BCL-2, FKHR, PAWR). These results taken together suggest that onset of gonadotropin surge triggers activation of various biological pathways and that the effects of growth factors and peptides on gonadotropin actions could be examined during preovulatory follicle development.

Hyper-activation of the TCP4 transcription factor in Arabidopsis thaliana accelerates multiple aspects of plant maturation

Plant organs are initiated as primordial outgrowths, and require controlled cell division and differentiation to achieve their final size and shape. Superimposed on this is another developmental program that orchestrates the switch from vegetative to reproductive to senescence stages in the life cycle. These require sequential function of heterochronic regulators. Little is known regarding the coordination between organ and organismal growth in plants. The TCP gene family encodes transcription factors that control diverse developmental traits, and a subgroup of class II TCP genes regulate leaf morphogenesis. Absence of these genes results in large, crinkly leaves due to excess division, mainly at margins. It has been suggested that these class II TCPs modulate the spatio-temporal control of differentiation in a growing leaf, rather than regulating cell proliferation per se. However, the link between class II TCP action and cell growth has not been established. As loss-of-function mutants of individual TCP genes in Arabidopsis are not very informative due to gene redundancy, we generated a transgenic line that expressed a hyper-activated form of TCP4 in its endogenous expression domain. This resulted in premature onset of maturation and decreased cell proliferation, leading to much smaller leaves, with cup-shaped lamina in extreme cases. Further, the transgenic line initiated leaves faster than wild-type and underwent precocious reproductive maturation due to a shortened adult vegetative phase. Early senescence and severe fertility defects were also observed. Thus, hyper-activation of TCP4 revealed its role in determining the timing of crucial developmental events, both at the organ and organism level.

Fatigue crack growth studies on lug joints

Pin loaded lug joints fitted with different types of pins are analysed in the presence of cracks at pin-plate interface. An algorithm for finite element contact stress analysis of joints developed earlier to deal with varying partial contact/separation at the pin-plate interface using a marching solution is used in the present analysis. Stress Intensity Factors (SIF) at the crack tips are evaluated using Modified Crack Closure Integral (MCCI) method within the realm of Linear Elastic Fracture Mechanics (LEFM) assumptions. A comparison of fatigue crack growth lives between interference and push fit pin joints is carried out using these SIF's. Results from a finite element analysis on a push fit pin joint are used to fit experimental fatigue crack growth data.

Growth Kinetics of Nanoclusters in Solution

We study the growth kinetics of nanoclusters in solution. There are two generic factors that drive growth: (a) reactions that produce the nanomaterial; and (b) diffusion of the nanomaterial due to chemical-potential gradients. We model the growth kinetics of ZnO nanoparticles via coupled dynamical equations for the relevant order parameters, We study this model both analytically and numerically. We find that there is a crossover in thenanocluster growth law: from L(t) similar to t(1/2) in the reaction-controlled regime to L(t) t(1/3) in the diffusion-controlled regime.