Mechanical characterization & regression analysis of Calamus rotang based hybrid natural fibre composite with findings reported on retrieval bending strength.

Research on Bio-based natural fiber material promoted the development of reinforcement and expand their possible structural applications. In this study, fibers are extracted from the stem of Calamus rotang (common rattan-Indian Species). Further, the fiber is processed to get novel hybrid combinations with glass fibers by manual hand lay-up technique. Three sets of samples were prepared for the different volume fractions of 60:40, 30:30:30, and 60:32:8 of glass fiber/epoxy as neat composite sample (NCS), a hybrid combination of C. rotang /glass fiber with epoxy as modified reinforced composite sample (MRCS) and glass fiber/epoxy with calamus stem powder as modified matrix composite sample (MMCS) respectively. Mechanical tests including tensile, flexural, impact, and ILSS tests are conducted as per ASTM Standards. Comparative studies have been done to evaluate the effect of novel species of C. rotang on mechanical properties with neat GFRP composites. Addition to this regression analysis has been carried out to achieve the experimental correlation for tensile and bending tests. Microstructural analysis for all the tested samples has been done to assess the fracture mode. Novel findings on retrieval bending strength for MMCS has been reported for the first time for composite materials. Study proves that novel species have a significant impact on the basic properties of materials.

Differential signaling through p190 and p210 BCR-ABL fusion proteins revealed by interactome and phosphoproteome analysis.

Two major types of leukemogenic BCR-ABL fusion proteins are p190BCR-ABLand p210BCR-ABL. Although the two fusion proteins are closely related, they can lead to different clinical outcomes. A thorough understanding of the signaling programs employed by these two fusion proteins is necessary to explain these clinical differences. We took an integrated approach by coupling protein-protein interaction analysis using biotinylation identification with global phosphorylation analysis to investigate the differences in signaling between these two fusion proteins. Our findings suggest that p190BCR-ABL and p210BCR-ABL differentially activate important signaling pathways, such as JAK-STAT, and engage with molecules that indicate interaction with different subcellular compartments. In the case of p210BCR-ABL, we observed an increased engagement of molecules active proximal to the membrane and in the case of p190BCR-ABL, an engagement of molecules of the cytoskeleton. These differences in signaling could underlie the distinct leukemogenic process induced by these two protein variants.

A short stereoselective synthesis of (+)-(6R,2'S)-cryptocaryalactone via ring-closing metathesis.

A short stereoselective synthesis of (+)-(6R,2'S)-cryptocaryalactone was successfully completed. Key steps included the application of Carreira's asymmetric alkynylation reaction to form a propargylic alcohol and subsequently lactone formation using the powerful ring-closing metathesis reaction.

Using molecular tethering to analyze the role of nuclear compartmentalization in the regulation of mammalian gene activity.

The mammalian nucleus has a complex structural organization that dynamically interacts with the genome. Chromatin is organized into discrete domains by association with distinct nuclear compartments enriched in structural and regulatory proteins. Growing evidence suggests that gene activity is modulated by interactions with these sub-nuclear compartments. Therefore, analyzing how nuclear architecture controls genome activity will be necessary to fully understand complex biological processes such as development and disease. In this article we describe a molecular methodology involving inducible tethering that can be used to position genes at the inner nuclear membrane (INM)-lamina compartment. The consequences of such directed re-positioning on gene activity or other DNA transactions can then be analyzed. This approach can be generalized and extended to position genes or chromosomal domains within other nuclear compartments thereby greatly facilitating the analysis of nuclear structure and its impact on genome activity.

Transcriptional repression mediated by repositioning of genes to the nuclear lamina.

Nuclear compartmentalization seems to have an important role in regulating metazoan genes. Although studies on immunoglobulin and other loci have shown a correlation between positioning at the nuclear lamina and gene repression, the functional consequences of this compartmentalization remain untested. We devised an approach for inducible tethering of genes to the inner nuclear membrane (INM), and tested the consequences of such repositioning on gene activity in mouse fibroblasts. Here, using three-dimensional DNA-immunoFISH, we demonstrate repositioning of chromosomal regions to the nuclear lamina that is dependent on breakdown and reformation of the nuclear envelope during mitosis. Moreover, tethering leads to the accumulation of lamin and INM proteins, but not to association with pericentromeric heterochromatin or nuclear pore complexes. Recruitment of genes to the INM can result in their transcriptional repression. Finally, we use targeted adenine methylation (DamID) to show that, as is the case for our model system, inactive immunoglobulin loci at the nuclear periphery are contacted by INM and lamina proteins. We propose that these molecular interactions may be used to compartmentalize and to limit the accessibility of immunoglobulin loci to transcription and recombination factors.

The Drosophila PAR domain protein 1 (Pdp1) gene encodes multiple differentially expressed mRNAs and proteins through the use of multiple enhancers and promoters.

Transcription factors are often expressed at several times and in multiple tissues during development and regulate diverse sets of downstream target genes by varying their combinatorial interactions with other transcription factors. The Drosophila Tropomyosin I (TmI) gene is regulated by a complex of proteins within the enhancer that synergistically interacts with MEF2 to activate TmI transcription as muscle cells fuse and differentiate. One of the components of this complex is PDP1 (PAR domain protein 1), a basic leucine zipper transcription factor that is highly homologous to three vertebrate genes that are members of the PAR domain subfamily. We have isolated and describe here the structure of the Pdp1 gene. The Pdp1 gene is complex, containing at least four transcriptional start sites and producing at least six different mRNAs and PDP1 isoforms. Five of the PDP1 isoforms differ by the substitution or insertion of amino acids at or near the N-terminal of the protein. At least three of these alternately spliced transcripts are differentially expressed in different tissues of the developing embryo in which PDP1 expression is correlated with the differentiation of different cell types. A sixth isoform is produced by splicing out part of the PAR and basic DNA binding domains, and DNA binding and transient transfection experiments suggest that it functions as a dominant negative inhibitor of transcription. Furthermore, two enhancers have been identified within the gene that express in the somatic mesodermal precursors to body wall muscles and fat body and together direct expression in other tissues that closely mimics that of the endogenous gene. These results show that Pdp1 is widely expressed, including in muscle, fat, and gut precursors, and is likely involved in the transcriptional control of different developmental pathways through the use of differentially expressed PDP1 isoforms. Furthermore, the similarities between Pdp1 and the other PAR domain genes suggest that Pdp1 is the homologue of the vertebrate genes.

Production and characterization of monoclonal antibodies for aflatoxin B1.

Hybridomas that secreted antibodies for aflatoxin B1 were selected using two immunization protocols referred to as A and B. Protocol A is a standard immunization method and resulted in the selection of only two clones that produced monoclonal antibodies against aflatoxin B1. In protocol B a unique immunization schedule which resulted in the generation of 10 hybridomas is described. Of the 10, one antibody was highly specific to B1, four antibodies reacted equally strongly with B1, G1 and weakly with B2. Another four reacted strongly with B1 and weakly with B2 and G1. One clone reacted equally strongly with B1, G1 and B2. Interestingly all the 10 antibodies showed little or no cross-reaction with G2.

Peanut yellow spot virus is a member of a new serogroup of Tospovirus genus based on small (S) RNA sequence and organization.

Peanut yellow spot virus (PYSV) represents a distinct tospovirus species based on serology and nucleic acid hybridization. THe sequence of the S RNA was 2970 nucleotides with 22 nucleotide long inverted repeats (with three mismatches) at the termini. The coding was ambisense with a long open reading frame (ORF) in each strand. The 5'-large ORF (1,400 nucleotides in the viral sense (v) strand) encoded a protein with a predicted size of 53.2 kDa that was identified as the nonstructural (NSs) protein based on 16-21% sequence identity and 42-48% sequence similarity with other tospoviruses. A 3' ORF (741 nucleotides) in the virus complementary (vc) sense encoded a 28.0 kDa protein that was identified as the nucleocapsid (N) gene based on immuno-blot analysis of the in vitro expressed protein with PYSV polyclonal antiserum. The predicted N protein had 24-28% amino acid sequence identity and 44-51% sequence similarity with the members of other serogroups. In contrast to other tospoviruses, a third ORF (204 nucleotides) occurred in the vc strand, which could encode a protein with a predicted size of 7.5 kDa with two strong hydrophobic regions. The low degree of homology of N and NSs protein sequences with other serogroup members coupled with an additional ORF suggests that PYSV should be classified as a distinct species of the Tospovirus genus. This conclusion also is supported by the absence of serological cross reaction with other serogroups, and biological characteristics including thrips transmission, symptoms and host range.

PDP1, a novel Drosophila PAR domain bZIP transcription factor expressed in developing mesoderm, endoderm and ectoderm, is a transcriptional regulator of somatic muscle genes.

In vertebrates, transcriptional control of skeletal muscle genes during differentiation is regulated by enhancers that direct the combinatorial binding and/or interaction of MEF2 and the bHLH MyoD family of myogenic factors. We have shown that Drosophila MEF2 plays a role similar to its vertebrate counterpart in the regulation of the Tropomyosin I gene in the development of Drosophila somatic muscles, however, unlike vertebrates, Drosophila MEF2 interacts with a muscle activator region that does not have binding sites for myogenic bHLH-like factors or any other known Drosophila transcription factors. We describe here the isolation and characterization of a component of the muscle activator region that we have named PDP1 (PAR domain protein 1). PDP1 is a novel transcription factor that is highly homologous to the PAR subfamily of mammalian bZIP transcription factors HLF, DBP and VBP/TEF. This is the first member of the PAR subfamily of bZIP transcription factors to be identified in Drosophila. We show that PDP1 is involved in regulating expression of the Tropomyosin I gene in somatic body-wall and pharyngeal muscles by binding to DNA sequences within the muscle activator that are required for activator function. Mutations that eliminate PDP1 binding eliminate muscle activator function and severely reduce expression of a muscle activator plus MEF2 mini-enhancer. These and previous results suggest that PDP1 may function as part of a larger protein/DNA complex that interacts with MEF2 to regulate transcription of Drosophila muscle genes. Furthermore, in addition to being expressed in the mesoderm that gives rise to the somatic muscles, PDP1 is also expressed in the mesodermal fat body, the developing midgut endoderm, the hindgut and Malpighian tubules, and the epidermis and central nervous system, suggesting that PDP1 is also involved in the terminal differentiation of these tissues.

Serum levels of IgA in peptic ulcers.

A study of the IgA levels in 43 duodenal ulcer (DU) patients and 8 gastric ulcer (GU) patients and their comparison with healthy controls reveals significantly elevated levels of IgA in DU and somewhat lower levels in GU. The levels were also associated with the genotypes of the patients for genetic markers such as ABO blood group, ABH sectetor status, haptoglobin, and alkaline phosphatase enzyme. Nutritional factors, such as vegetarianism, chili consumption, and habits such as smoking and alcoholism also showed variation in the IgA levels. These results indicate the response and role of IgA in the immunological mechanisms involving mucosal protection and autoimmunity in ulceration processes in the stomach.