MOLECULAR ANALYSIS OF STREPTOCOCCUS THERMOPHILUS SUPPORTING PROBIOTIC ACTION TOWARDS DRUG TARGETS

Streptococcus thermophilus is a Gram-positive facultative anaerobe with probiotic action. Nearly 2070 genes arepredicted in complete genome out of which 566 genes are related to hypothetical and uncharacterized genes. Thepredicted gene percent obtained by characterization of complete genome of S.thermophilus is 72.67% with 25functional categories. There is high number of transport, protein synthesis, binding and cell envelop proteins thatdon’t have pathogen responses. The genes ptsK, glgP, malQ, lacZ, amyL, scrB, etc are related to energy metabolism(with biodegradation and synthesis of polysaccharides) are found in chromosome CNRZ 1066. The genes such asglgP, malQ, lacZ and amyl had shown direct interactions that has not found direct link with scrB gene. All theproteins are observed with more the 50 percent identities that are present in homology models

PROCESS MODELING FOR FLUID-INTERFACE SUPPORTED RESIN PRINTING

The article details the latest design and implementation of a stereolithography (SLA) based technique, Fluid Interface Supported Printing (FISP). The FISP technique involves printing from a thin resin layer above a static, immiscible support fluid. The support fluid prevents deflection of overhanging geometry by providing a buoyant force equal to that of the gravitational force due to the minute density difference between the support fluid and cured resin. Complex curing and shrinkage dynamics are a primary knowledge gap. A COMSOL multiphysics simulation model was developed to simulate the curing process, including volumetric light intensity and optimized parameters for chemical reaction kinetics. The article presents a pathway for further enhancing and validating the simulation model.Mechanical Engineerin

Identification and Characterization of Nucleolin as a COUP-TFII Coactivator of Retinoic Acid Receptor β Transcription in Breast Cancer Cells

The orphan nuclear receptor COUP-TFII plays an undefined role in breast cancer. Previously we reported lower COUP-TFII expression in tamoxifen/endocrine-resistant versus sensitive breast cancer cell lines. The identification of COUP-TFII-interacting proteins will help to elucidate its mechanism of action as a transcriptional regulator in breast cancer.FLAG-affinity purification and multidimensional protein identification technology (MudPIT) identified nucleolin among the proteins interacting with COUP-TFII in MCF-7 tamoxifen-sensitive breast cancer cells. Interaction of COUP-TFII and nucleolin was confirmed by coimmunoprecipitation of endogenous proteins in MCF-7 and T47D breast cancer cells. In vitro studies revealed that COUP-TFII interacts with the C-terminal arginine-glycine repeat (RGG) domain of nucleolin. Functional interaction between COUP-TFII and nucleolin was indicated by studies showing that siRNA knockdown of nucleolin and an oligonucleotide aptamer that targets nucleolin, AS1411, inhibited endogenous COUP-TFII-stimulated RARB2 expression in MCF-7 and T47D cells. Chromatin immunoprecipitation revealed COUP-TFII occupancy of the RARB2 promoter was increased by all-trans retinoic acid (atRA). RARβ2 regulated gene RRIG1 was increased by atRA and COUP-TFII transfection and inhibited by siCOUP-TFII. Immunohistochemical staining of breast tumor microarrays showed nuclear COUP-TFII and nucleolin staining was correlated in invasive ductal carcinomas. COUP-TFII staining correlated with ERα, SRC-1, AIB1, Pea3, MMP2, and phospho-Src and was reduced with increased tumor grade.Our data indicate that nucleolin plays a coregulatory role in transcriptional regulation of the tumor suppressor RARB2 by COUP-TFII

Use of multiple time points to model parotid differentiation

In order to understand the process of terminal differentiation in salivary acinar cells, mRNA and microRNA expression was measured across the month long process of differentiation in the parotid gland of the rat. Acinar cells were isolated at either nine time points (mRNA) or four time points (microRNA) in triplicate using laser capture microdissection (LCM). One of the values of this dataset comes from the high quality RNA (RIN > 7) that was used in this study, which can be prohibitively difficult to obtain from such an RNaseI-rich tissue. Global mRNA expression was measured by rat genome microarray hybridization (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE65586), and expression of microRNAs by qPCR array (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE65324). Comparing expression at different ages, 2656 mRNAs and 64 microRNAs were identified as differentially expressed. Because mRNA expression was sampled at many time points, clustering and regression analysis were able to identify dynamic expression patterns that had not been implicated in acinar differentiation before. Integration of the two datasets allowed the identification of microRNA target genes, and a gene regulatory network. Bioinformatics R code and additional details of experimental methods and data analysis are provided

Mechanical, Microstructural and Drying Shrinkage Properties of NaOH-Pretreated Crumb Rubber Concrete: RSM-Based Modeling and Optimization

One of the primary causes of the low mechanical properties of rubberized concrete is the weak bond between crumb rubber (CR) and hardened cement paste. Many CR pretreatment techniques have been researched in an attempt to mitigate this problem. The NaOH pretreatment method is one of the most widely used, although the reported results are inconsistent due to the absence of standardized NaOH pretreatment concentrations and CR replacement levels. This study aims to develop models for predicting the mechanical and shrinkage properties of NaOH-pretreated CR concrete (NaOH-CRC) and conduct multi-objective optimization using response surface methodology (RSM). The RSM generated experimental runs using three levels (0, 5, and 10%) of both NaOH pretreatment concentration and the CR replacement level of fine aggregate by volume as the input factors. At 28 days, the concrete’s compressive, flexural, and tensile strengths (CS, FS, and TS), as well as its drying shrinkage (S), were evaluated as the responses. The results revealed that higher CR replacements led to lower mechanical strengths and higher shrinkage. However, the strength loss and the shrinkage significantly reduced by 22%, 44%, 43%, and 60% for CS, FS, TS, and S, respectively, after the pretreatment. Using field-emission scanning electron microscopy (FESEM), the microstructural investigation indicated a significantly reduced interfacial transition zone (ITZ) with increasing NaOH pretreatment. The developed RSM models were evaluated using ANOVA and found to have high R2 values ranging from 78.7% to 98%. The optimization produced NaOH and CR levels of 10% and 2%, respectively, with high desirability of 71.4%

Rapid microwave-assisted biosynthesis of chitooligosaccharide coated silver nanoparticles: Assessments of antimicrobial activity for paediatric pulp therapy

Silver nanoparticles (AgNPs) have gained much attention in the treatment of dental caries due to their excellent antimicrobial activities. In this study, we have biosynthesised chitooligosaccharide coated silver nanoparticles (COS-AgNPs) using a rapid microwave-assisted method. The physical and chemical characterisations of biosynthesised COS-AgNPs were investigated using UV–visible spectroscopy (UV–vis), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR), X-Ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) equipped with selected area electron diffraction (SAED). Further, the developed COS-AgNPs were assessed for antimicrobial activity on Streptococcus mutans. From the DLS analysis, the average particle size of the COS-AgNPs was found to be ∼32 nm. The morphologies of the COS-AgNPs were monodisperse and spherical in shape. XRD peaks are coherent with the standard peaks of AgNPs. Antimicrobial activity of COS-AgNPs showed a significant zone of inhibition due to the synergistic effect of COS-AgNPs. Furthermore, the antibacterial potential of developed COS-AgNPs-Ca(OH)2 paste was checked against Streptococcus mutans, Staphylococcus aureus, and Enterococcus faecalis. COS-AgNPs based composite paste showed better inhibition zone against the tested microbes. Hence, COS-AgNPs and COS-AgNPs-Ca(OH)2 based paste materials will be promising biomaterials for pulp therapy in paediatric dentistry.The work is supported by Yenepoya Dental College, Yenepoya (Deemed to be University), Mangalore, Karnataka.Scopu