Contribution of Environmental Constituents in the Genomic Disruption of Cytokeratins

Cytokeratins are keratinous protein and assist cells to reduce mechanical stress on the intracytoplasmic layer of epithelial tissue. There are several unspecified mutations in the epithelial layer that may induces by environmental mutagens and pathogens. The unspecified mutations in the epithelium surface also disrupt biology of skin at multiple different levels and cause innate keratinizing disorders. These serve as a root generator of neurohormones and neuropeptides which mainly partake in the disruption. Generally, all 54 unique genes of human keratin partake in mutations and cause cutaneous tissue fragility, skin hypertrophic, and malignant transformation. In this chapter, unspecific factors that involved in the pathogenesis of skin diseases and the ways by which such keratin changes might harness to alleviate different skin conditions are also included. Consequently, the contribution of environmental changes in the frontier of mutations or misregulations of the cytokeratin genes, is also cited here

Peptide aptamer-modified single-walled carbon nanotube-based transistors for high-performance biosensors

Biosensors employing single-walled carbon nanotube field-effect transistors (SWCNT FETs) offer ultimate sensitivity. However, besides the sensitivity, a high selectivity is critically important to distinguish the true signal from interference signals in a non-controlled environment. This work presents the first demonstration of the successful integration of a novel peptide aptamer with a liquid-gated SWCNT FET to achieve highly sensitive and specific detection of Cathepsin E (CatE), a useful prognostic biomarker for cancer diagnosis. Novel peptide aptamers that specifically recognize CatE are engineered by systemic in vitro evolution. The SWCNTs were firstly grown using the thermal chemical vapor deposition (CVD) method and then were employed as a channel to fabricate a SWCNT FET device. Next, the SWCNTs were functionalized by noncovalent immobilization of the peptide aptamer using 1-pyrenebutanoic acid succinimidyl ester (PBASE) linker. The resulting FET sensors exhibited a high selectivity (no response to bovine serum albumin and cathepsin K) and label-free detection of CatE at unprecedentedly low concentrations in both phosphate-buffered saline (2.3 pM) and human serum (0.23 nM). Our results highlight the use of peptide aptamer-modified SWCNT FET sensors as a promising platform for near-patient testing and point-of-care testing applications

CRISPR-Cas9: Role in Processing of Modular Metabolic Engineered Bio-Based Products

Biogenetic engineering is a significant technology to sensibly manage microbial metabolic product factories. Genome modification methods for efficiently controlling and modifying genes at the genome level have progressed in biogenetic engineering during the last decade. CRISPR is genome editing technology that allows for the modification of organisms’ genomes. CRISPR and its related RNA-guided endonuclease are versatile advanced immune system frameworks for defending against foreign DNA and RNAs. CRISPR is efficient, accessible, and trustworthy genomic modification tool in unparalleled resolution. At present, CRISPR-Cas9 method is expanded to industrially manipulate cells. Metabolically modified organisms are quickly becoming interested in the production of different bio-based components. Here, chapter explore about the control productivity of targeted biomolecules in divergent cells based on the use of different CRISPR-related Cas9

Solid-phase translation and RNA–protein fusion: a novel approach for folding quality control and direct immobilization of proteins using anchored mRNA

A novel cell-free translation system is described in which template-mRNA molecules were captured onto solid surfaces to simultaneously synthesize and immobilize proteins in a more native-state form. This technology comprises a novel solid-phase approach to cell-free translation and RNA–protein fusion techniques. A newly constructed biotinylated linker-DNA which enables puromycin-assisted RNA–protein fusion is ligated to the 3′ ends of the mRNA molecules to attach the mRNA-template on a streptavidin-coated surface and further to enable the subsequent reactions of translation and RNA–protein fusion on surface. The protein products are therefore directly immobilized onto solid surfaces and furthermore were discovered to adopt a more native state with proper protein folding and superior biological activity compared with conventional liquid-phase approaches. We further validate this approach via the production of immobilized green fluorescent protein (GFP) on microbeads and by the production and assay of aldehyde reductase (ALR) enzyme with 4-fold or more activity. The approach developed in this study may enable to embrace the concept of the transformation of ‘RNA chip-to-protein chip’ using a solid-phase cell-free translation system and thus to the development of high-throughput microarray platform in the field of functional genomics and in vitro evolution

Towards the synthesis of alpha-cyclo peptide nucleic acid monomers

Peptide nucleic acids (PNAs) are potentially useful as mimics of DNA for gene therapy treatment of cancer and other genetic disorders. The original PNA molecule developed by Nielsen et al. lacks selectivity towards nucleic acid targets (DNA vs RNA) and the ability to incorporate new monomers for each alteration to be examined. Our design of L-α-PNA allows for the incorporation of different appropriate α-amino acid spacers into the oligomers during peptide synthesis. Selectivity towards nucleic acid targets is improved by freezing out conformation with incorporating cyclopentane ring in L-α-PNA. The aim of the research reported in this thesis was to develop viable synthetic routes to α-cycloPNA monomers, constrained analogues of PNA monomers. Our initial objective was to optimize an established synthetic pathway to a key cyclopentane α–amino acid intermediate suitably functionalized at the 3-position to enable subsequent preparation of pyrimidine and purine α-cycloPNA monomers. However, reproduction of the key cyclization step in our prototypical route to a key 3-alkoxy-1-aminocyclopentanecaboxylate precursor proved to be problematic and so alternative synthetic pathways were explored. All four diastereoisomers (203, 143, 204 and 144) of the required cyclopentane α–amino acid intermediate were prepared via hydroboration of ethyl N-Boc-1-aminocyclopent-3-ene-1-carboxylic acid in overall yields of 6.2% for the (1S/R, 3S/R)-alcohols (204 and 144) diastereoisomeric mixture and 13% for the (1R/S, 3S/R)-alcohol (203 and 143), also isolated as an inseparable diastereoisomeric mixture. Unfortunately, all attempts to resolve these mixtures failed. With both the (1S/R, 3S/R)-alcohols and (1R/S, 3S/R)-alcohols to hand, synthesis of a cytosine derivative of the α-cycloPNA monomer was investigated. The mixed (1R/S, 3S/R)-alcohol diastereoisomers were first converted into their respective camphorsulfonate derivatives (232 and 233) and treated with Cbz-protected cytosine. Unexpectedly, the corresponding O2-alkylated cytosine α-cycloPNA monomers (266 and 267) were afforded. Asymmetric hydroboration of the chiral cyclopentene intermediate bearing D-menthyl ester using either (+)- or (-)-IpcBH2.TMEDA were studied. This also gave unresolvable diastereoisomeric mixtures of (1R/S, 3S/R)-alcohols (299 and 300), in 82% yield, and (1R/S, 3S/R)-alcohols (299 and 300), in 69% yield. Finally, treatment of the (1R/S, 3S/R)-brosylate mixture derived from the (1R/S, 3S/R)-alcohols of D-menthyl esters with Cbz-protected cytosine again yielded the O2-alkylated cytosine α-cycloPNA monomers

Microintaglio Printing for Soft Lithography-Based in Situ Microarrays

Advances in lithographic approaches to fabricating bio-microarrays have been extensively explored over the last two decades. However, the need for pattern flexibility, a high density, a high resolution, affordability and on-demand fabrication is promoting the development of unconventional routes for microarray fabrication. This review highlights the development and uses of a new molecular lithography approach, called “microintaglio printing technology”, for large-scale bio-microarray fabrication using a microreactor array (µRA)-based chip consisting of uniformly-arranged, femtoliter-size µRA molds. In this method, a single-molecule-amplified DNA microarray pattern is self-assembled onto a µRA mold and subsequently converted into a messenger RNA or protein microarray pattern by simultaneously producing and transferring (immobilizing) a messenger RNA or a protein from a µRA mold to a glass surface. Microintaglio printing allows the self-assembly and patterning of in situ-synthesized biomolecules into high-density (kilo-giga-density), ordered arrays on a chip surface with µm-order precision. This holistic aim, which is difficult to achieve using conventional printing and microarray approaches, is expected to revolutionize and reshape proteomics. This review is not written comprehensively, but rather substantively, highlighting the versatility of microintaglio printing for developing a prerequisite platform for microarray technology for the postgenomic era

DEP-On-Go for Simultaneous Sensing of Multiple Heavy Metals Pollutants in Environmental Samples

We describe a simple and affordable “Disposable electrode printed (DEP)-On-Go” sensing platform for the rapid on-site monitoring of trace heavy metal pollutants in environmental samples for early warning by developing a mobile electrochemical device composed of palm-sized potentiostat and disposable unmodified screen-printed electrode chips. We present the analytical performance of our device for the sensitive detection of major heavy metal ions, namely, mercury, cadmium, lead, arsenic, zinc, and copper with detection limits of 1.5, 2.6, 4.0, 5.0, 14.4, and, 15.5 μg·L−1, respectively. Importantly, the utility of this device is extended to detect multiple heavy metals simultaneously with well-defined voltammograms and similar sensitivity. Finally, “DEP-On-Go” was successfully applied to detect heavy metals in real environmental samples from groundwater, tap water, house dust, soil, and industry-processed rice and noodle foods. We evaluated the efficiency of this system with a linear correlation through inductively coupled plasma mass spectrometry, and the results suggested that this system can be reliable for on-site screening purposes. On-field applications using real samples of groundwater for drinking in the northern parts of India support the easy-to-detect, low-cost (<1 USD), rapid (within 5 min), and reliable detection limit (ppb levels) performance of our device for the on-site detection and monitoring of multiple heavy metals in resource-limited settings

Arsenic exposure and perception of health risk due to groundwater contamination in Majuli (river island), Assam, India

Island populations are rarely studied for risk of arsenic (As) poisoning. As poisoning, multimetal contamination and people�s perceptions of health risks were assessed on India�s Majuli Island, the largest inhabited river island in the world. This holistic approach illustrated the association of groundwater contamination status with consequent health risk by measuring levels of inorganic arsenic (iAs) in groundwater, borehole sediment and biological samples (hair, nails and urine). Piper and Gibbs�s plots discerned the underlying hydrogeochemical processes in the aquifer. Demographic data and qualitative factors were evaluated to assess the risks and uncertainties of exposure. The results exhibited significant enrichment of groundwater with As, Mn and Fe along with significant body burden. Maximum Hazard Index values indicated severe non-carcinogenic health impacts as well as a significantly elevated risk of cancer for both adults and children. Most (99%) of the locally affected population did not know about the adverse health impacts of metal contamination, and only 15% understood bodily ailments and health issues. Various aspects of the island environment were used to elucidate the status of contamination and future risk of disease. A projection showed adverse health outcomes rising significantly, especially among the young population of Majuli, due to overexposure to not only As but also Ba, Mn and Fe.by Ritusmita Goswami, Manish Kumar, Nivedita Biyani and Patrick J. She