Single-Circulating Tumor Cell Whole Genome Amplification to Unravel Cancer Heterogeneity and Actionable Biomarkers

The field of single-cell analysis has advanced rapidly in the last decade and is providing new insights into the characterization of intercellular genetic heterogeneity and complexity, especially in human cancer. In this regard, analyzing single circulating tumor cells (CTCs) is becoming particularly attractive due to the easy access to CTCs from simple blood samples called “liquid biopsies”. Analysis of multiple single CTCs has the potential to allow the identification and characterization of cancer heterogeneity to guide best therapy and predict therapeutic response. However, single-CTC analysis is restricted by the low amounts of DNA in a single cell genome. Whole genome amplification (WGA) techniques have emerged as a key step, enabling single-cell downstream molecular analysis. Here, we provide an overview of recent advances in WGA and their applications in the genetic analysis of single CTCs, along with prospective views towards clinical applications. First, we focus on the technical challenges of isolating and recovering single CTCs and then explore different WGA methodologies and recent developments which have been utilized to amplify single cell genomes for further downstream analysis. Lastly, we list a portfolio of CTC studies which employ WGA and single-cell analysis for genetic heterogeneity and biomarker detection.     

The Properties of Synthetic Amylopectin with Long External-Chains

Samples of synthetic amylopectins with long external chain-lengths have been prepared from degraded, waxy maize amylopectin and glucose-1-phosphate by the action of potato phosphorylase. Samples having external chain-lengths determined by enzymic assay of 23.4, 29.9, 46.5, and 109.5 glucose units (original sample = 13.7 glucose units) were prepared. The iodine-staining properties of these materials have been investigated, and a relation between λ_max and average chain-length evaluated. A theoretical relation between chain-length and β-amylolysis limit has been proposed. The iodine-binding capacity of the samples has been studied in detail by potentiometric titration at 1.5 °C and 20 °C. Unambiguous conclusions regarding the observed iodine binding capacity and the length of external chain have been made for the first time. The properties of the synthetic products are discussed.     

Synthesis and Anti-Cancer Activity of New Pyrazolinyl-Indole Derivatives: Pharmacophoric Interactions and Docking Studies for Identifying New EGFR Inhibitors

Newly designed series of indole-containing pyrazole analogs, pyrazolinylindoles, were synthesized, and their structures were confirmed based on the spectral data of the ¹H NMR, ¹³C NMR, and HR-MS analyses. Preliminary anti-cancer activity testings were carried out by the National Cancer Institute, United States of America (NCI, USA). Compounds HD02, HD05, and HD12 demonstrated remarkable cytotoxic activities against nine categories of cancer types based cell line panels which included leukemia, colon, breast, melanoma, lungs, renal, prostate, CNS, and ovarian cancer cell lines. The highest cytotoxic effects were exhibited by the compounds HD02 [1-(5-(1-H-indol-3-yl)-3-(p-tolyl)-4,5-dihydro-1H-pyrazol-1-yl)-2-phenylethanone], HD05 [1-(3-(4-chlorophenyl)-5-(1H-indol-3-yl)-4,5-dihydro-1H-pyrazol-1-yl)-2-phenoxyethanone], and HD12 [(3-(4-chlorophenyl)-5-(1H-indol-3-yl)-4,5-dihydro-1H-pyrazol-1-yl)(pyridin-4-yl)methanone] against some of the 56 types of NCI-based cell lines in different panels. Compound HD05 showed the maximum range of cancer cell growth inhibitions against all categories of the cell lines in all nine panels. On average, in comparison to the referral standard, imatinib, at a dose level of 10 µM, the HD05 showed significant activity against leukemia in the range of 78.76%, as compared to the imatinib at 9% of cancer cells’ growth inhibitions. Molecular docking simulation studies were performed in silico on the epidermal growth factor receptor (EGFR) tyrosine kinase, in order to validate the activity.     

Influence of Y~(3+),Yb~(3+),Gd~(3+) cations on structural and electromagnetic properties of CuFe_2O_4 nanoferrites prepared via one step sol-gel method

Rare earths(REs) play a key role in distorting spinel structure by creating some defects at the lattice sites and make them suitable for magnetodielectric applications.In the present study,the nanoferrites of CuRE_(0.02)Fe_(1.98)O_4,where REs=Y~(3+),Yb~(3+),Gd~(3+),were prepared using one step sol-gel method.The prepared samples are copper ferrite(CFO),yttrium doped copper ferrite(Y-CFO),ytterbium doped copper ferrite(Yb-CFO) and gadolinium doped copper ferrite(Gd-CFO),respectively.The single-phase structure of all the REs doped nanoferrites was determined by X-ray diffraction(XRD) analysis.The porosity,agglomerations and grain size of the REs doped copper ferrite were examined using field emission scanning electron microscopy(FESEM) analysis.Fourier transform infrared spectroscopy(FTIR)elaborates the phase formation and environmental effects on the REs doped nanoparticles(NPs).The recorded room temperature M-H loops from a vibrating sample magnetometer(VSM) elucidate the magnetic properties of the REs doped spinel nanoferrites.The magnetic saturation(M_s) was calculated in the range of 23.08 to 51.78 emu/g.The calculated coercivity values(272.6 to 705.60 Oe) confirm the soft magnetic behavior of REs doped copper ferrites.Furthermore,the electromagnetic and dielectric properties were assessed using a Vector network analyzer(VNA) from 1 to 6 GHz.The permeability,permittivity,dielectric tangent loss and electric modulus of the REs doped spinel ferrites illustrate that the prepared NPs may be suitable for microwave and high frequency applications.     

Mini-maximizing two qubit quantum computations

Two qubit quantum computations are viewed as two player, strictly competitive games and a game-theoretic measure of optimality of these computations is developed. To this end, the geometry of Hilbert space of quantum computations is used to establish the equivalence of game-theoretic solution concepts of Nash equilibrium and mini-max outcomes in games of this type, and quantum mechanisms are designed for realizing these mini-max outcomes.     

Crypt analysis of two time pads in case of compressed speech

Keystream reuse, also known as the two time pad problem, is a well known weakness in stream ciphers. The implementers of the cryptographic algorithms are still underestimating this threat. The keystream reuse exploitation techniques presented so far assume the underlying plaintext to be textual data and all the heuristics presented previously are based on the language characteristics of the underlying text based data, which fail when compression is applied on the plaintext before encryption. This paper presents exploitation techniques for two time pads in case of stream ciphered digitized and compressed speech signals. In this paper we show that how an adversary can automatically recover the digitized speech signals encrypted under the same keystream provided the language (e.g. English) and digital encoding/compression scheme details of the underlying speech signals are known. Our technique of cryptanalysis is based on the modeling of the speech parameters by exploiting the inter frame correlations between each components of the speech vectors in different frames and then using these models to decode the two speech signals in the keystream reuse scenario. The technique is flexible enough to incorporate all modern speech coding schemes based on parameter or hybrid encoding and compression techniques. The simulation experiments have showed promising results for most of the present day speech digitization and compression techniques.     

Delivering nanotechnology to the healthcare, IT and environmental sectors — A perspective from the ‘London centre for nanotechnology’

Nanotechnology research is booming worldwide, having an impact on multiple sectors and with a general belief that medical and biological applications will form the greatest area of expansion over the next decade, driven by an attempt to bring radical solutions to areas of unmet medical need. What is true in the USA is also being fulfilled in Europe, though generally at a significantly lower investment level, even for ’large’ capital infrastructure and interdisciplinary centres. Against this, the UK and its European partners are following the maxim ’small is beautiful’ and are attempting to identify and grow academic research and commercial businesses in areas that traditional nanotechnology developments out of engineering or physics find challenging. Thus, University College London and Imperial College, in a major joint project linked to other centres of excellence both in the UK and in the rest of Europe, are building upon their internationally competitive medical and hard-matter research activities to focus on and develop nanotechnology as a major sector of research activity. The two universities together form one of the largest centres of biomedical research outside the USA. A novel approach to commercialisation has been taken, exemplified by the establishment with government and private equity funds of a ‘Bionanotechnology Centre’ — this will act as a portal for UK industry to access specialist skills to solve issues relating to developing nanotechnology-based medical applications, for example, for environmental screening, diagnostics and therapy. This paper reviews our academic and business strategy with examples from our current research portfolio, biased towards medicine as the London Centre for Nanotechnology’s most rapidly growing area.     

SCM: Secure and accountable TLS certificate management

In classical public‐key infrastructure (PKI), the certificate authorities (CAs) are fully trusted, and the security of the PKI relies on the trustworthiness of the CAs. However, recent failures and compromises of CAs showed that if a CA is corrupted, fake certificates may be issued, and the security of clients will be at risk. As emerging solutions, blockchain‐ and log‐based PKI proposals potentially solved the shortcomings of the PKI, in particular, eliminating the weakest link security and providing a rapid remedy to CAs' problems. Nevertheless, log‐based PKIs are still exposed to split‐world attacks if the attacker is capable of presenting two distinct signed versions of the log to the targeted victim(s), while the blockchain‐based PKIs have scaling and high‐cost issues to be overcome. To address these problems, this paper presents a secure and accountable transport layer security (TLS) certificate management (SCM), which is a next‐generation PKI framework. It combines the two emerging architectures, introducing novel mechanisms, and makes CAs and log servers accountable to domain owners. In SCM, CA‐signed domain certificates are stored in log servers, while the management of CAs and log servers is handed over to a group of domain owners, which is conducted on the blockchain platform. Different from existing blockchain‐based PKI proposals, SCM decreases the storage cost of blockchain from several hundreds of GB to only hundreds of megabytes. Finally, we analyze the security and performance of SCM and compare SCM with previous blockchain‐ and log‐based PKI schemes.