Screening of Hibiscus and Cinnamomum Plants and Identification of Major Phytometabolites in Potential Plant Extracts Responsible for Apoptosis Induction in Skin Melanoma and Lung Adenocarcinoma Cells

Carcinogenesis is a major concern that severely affects the human population. Owing to persistent demand for novel therapies to treat and prohibit this lethal disease, research interest among scientists is drawing its huge focus toward natural products, as they have minimum toxicity comparable with existing treatment methods. The plants produce secondary metabolites, which are known to have the anticancer potential for clinical drug development. Furthermore, the use of nanocarriers could boost the solubility and stability of phytocompounds to obtain site-targeting delivery. The identification of potential phytochemicals in natural compounds would be beneficial for the synthesis of biocompatible nanoemulsions. The present study aimed to investigate the potential cytotoxicity of ethanol extracts of Hibiscus syriacus and Cinnamomum loureirii Nees plant parts on human skin melanoma (G361) and lung adenocarcinoma (A549) cells. Importantly, biochemical analysis results showed the presence of high phenol (50–55 µgGAE/mg) and flavonoids [42–45 µg quercetin equivalents (QE)/mg] contents with good antioxidant activity (40–58%) in C. loureirii Nees plants extracts. This plant possesses potent antiproliferative activity (60–90%) on the malignant G361 and A549 and cell lines correlated with the production of nitric oxide. Especially, C. loureirii plant extracts have major metabolites that exhibit cancer cell death associated with cell cycle arrest. These findings support the potential application of Cinnamomum for the development of therapeutic nanoemulsion in future cancer therapy.

Self-Assembly of DNA Molecules: Towards DNA Nanorobots for Biomedical Applications

DNA nanotechnology takes DNA molecule out of its biological context to build nanostructures that have entered the realm of robots and thus added a dimension to cyborg and bionic systems. Spurred by spring-like properties of DNA molecule, the assembled nanorobots can be tuned to enable restricted, mechanical motion by deliberate design. DNA nanorobots can be programmed with a combination of several unique features, such as tissue penetration, site-targeting, stimuli responsiveness, and cargo-loading, which makes them ideal candidates as biomedical robots for precision medicine. Even though DNA nanorobots are capable of detecting target molecule and determining cell fate via a variety of DNA-based interactions both in vitro and in vivo, major obstacles remain on the path to real-world applications of DNA nanorobots. Control over nanorobot’s stability, cargo loading and release, analyte binding, and dynamic switching both independently and simultaneously represents the most eminent challenge that biomedical DNA nanorobots currently face. Meanwhile, scaling up DNA nanorobots with low-cost under CMC and GMP standards represents other pertinent challenges regarding the clinical translation. Nevertheless, DNA nanorobots will undoubtedly be a powerful toolbox to improve human health once those remained challenges are addressed by using a scalable and cost-efficient method.

The Efficiency of AuNPs in Cancer Cell Targeting Compared to Other Nanomedicine Technologies Using Fuzzy PROMETHEE

Cancer is a disease with rare, diverse symptoms, causing abnormal cell growth in an uncontrolled way, leading to cell damage, apoptosis, and eventually death of the patient. This study uses the Fuzzy PROMETHEE technique to develop a new path for cancer treatment based on nanoparticles (NPs) applications, used in controlled anticancer drug delivery (drug release, toxicity, and unspecific site targeting) to enhance patient safety. The different nanoparticles employed in the drug delivery analysis are gold nanoparticles (AuNPs), liposomes, dendrimers, polymeric micelles (PMs), and quantum dots (QDs). Fuzzy predictable preference organization mode and evaluation multicriteria choice were used as tactics in making the best decision using the data from the factors of cost, size, shape, surface charge, ligand type, pH and temperature stimuli, biocompatibility, accumulation ratio, toxicity, specificity, stability, efficacy, adverse effect, and safety factor of the NPs. The results obtained from the total net flow of the visual PROMETHEE scenario for anticancer drug delivery, based on NPs data analysis, show that AuNPs are ranked the highest among the other NPs. The Phi values obtained for the NPs are as follows: AuNPs (0.1428), PMs (0.0280), QDs (−0.0467), dendrimers (−0.0593), and liposomes (−0.0649). This study highlights the optimal choice of NPs as an intelligent drug delivery system that facilitates therapeutic efficiency, where cancer cells are accurately targeted to enhance treatment quality and patient safety.

Justify my love: Cognitive dissonance reduction among perpetrators of online and offline infidelity

A longitudinal survey study was conducted to examine which strategies for reducing cognitive dissonance were used among men engaging in infidelity. Data were collected in two waves, 1 month apart ( n time1 = 1514, n time2 = 425), from a sample of male users of Ashley Madison, a “married dating” site targeting users who are seeking to engage in infidelity. Because perpetrators of infidelity may justify their behaviors differently depending on whether they cheated in an online environment, both online and offline infidelity behaviors were considered. Results indicated that attitude change and self-concept change were positively related to online infidelity, while only self-concept change was positively related to offline infidelity, suggesting their differential effectiveness for various communication media. Self-concept change, attitude change, and denial of responsibility were negatively related to psychological discomfort and perceived negative impact at time 2, indicating their relative success for reducing negative psychological outcomes compared to other strategies such as adding consonant cognitions.

Exon-Skipping Antisense Oligonucleotides for Cystic Fibrosis Therapy

Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene cause cystic fibrosis (CF), and the CFTR-W1282X nonsense mutation causes a severe form of CF. Although Trikafta and other CFTR-modulation therapies benefit most CF patients, targeted therapy for patients with the W1282X mutation is lacking. The CFTR-W1282X protein has residual activity, but is expressed at a very low level due to nonsense-mediated mRNA decay (NMD). NMD-suppression therapy and read-through therapy are actively being researched for CFTR nonsense mutants. NMD suppression could increase the mutant CFTR mRNA, and read-through therapies may increase the levels of full-length CFTR protein. However, these approaches have limitations and potential side effects: because the NMD machinery also regulates the expression of many normal mRNAs, broad inhibition of the pathway is not desirable; and read-through drugs are inefficient, partly because the mutant mRNA template is subject to NMD. To bypass these issues, we pursued an exon-skipping antisense oligonucleotide (ASO) strategy to achieve gene-specific NMD evasion. A cocktail of two splice-site-targeting ASOs induced the expression of CFTR mRNA without the PTC-containing exon 23 (CFTR-Δex23), which is an in-frame exon. Treatment of human bronchial epithelial cells with this cocktail of ASOs that target the splice sites flanking exon 23 results in efficient skipping of exon 23 and an increase in CFTR-Δex23 protein. The splice-switching ASO cocktail increases the CFTR-mediated chloride current in human bronchial epithelial cells. Our results set the stage for developing an allele-specific therapy for CF caused by the W1282X mutation.

HIV Capsid and Integration Targeting

Integration of retroviral reverse transcripts into the chromosomes of the cells that they infect is required for efficient viral gene expression and the inheritance of viral genomes to daughter cells. Before integration can occur, retroviral reverse transcription complexes (RTCs) must access the nuclear environment where the chromosomes reside. Retroviral integration is non-random, with different types of virus-host interactions impacting where in the host chromatin integration takes place. Lentiviruses such as HIV efficiently infect interphase cells because their RTCs have evolved to usurp cellular nuclear import transport mechanisms, and research over the past decade has revealed specific interactions between the HIV capsid protein and nucleoporin (Nup) proteins such as Nup358 and Nup153. The interaction of HIV capsid with cleavage and polyadenylation specificity factor 6 (CPSF6), which is a component of the cellular cleavage and polyadenylation complex, helps to dictate nuclear import as well as post-nuclear RTC invasion. In the absence of the capsid-CPSF6 interaction, RTCs are precluded from reaching nuclear speckles and gene-rich regions of chromatin known as speckle-associated domains, and instead mis-target lamina-associated domains out at the nuclear periphery. Highlighting this area of research, small molecules that inhibit capsid-host interactions important for integration site targeting are highly potent antiviral compounds.

Factors that mold the nuclear landscape of HIV-1 integration

The integration of retroviral reverse transcripts into the chromatin of the cells that they infect is required for virus replication. Retroviral integration has far-reaching consequences, from perpetuating deadly human diseases to molding metazoan evolution. The lentivirus human immunodeficiency virus 1 (HIV-1), which is the causative agent of the AIDS pandemic, efficiently infects interphase cells due to the active nuclear import of its preintegration complex (PIC). To enable integration, the PIC must navigate the densely-packed nuclear environment where the genome is organized into different chromatin states of varying accessibility in accordance with cellular needs. The HIV-1 capsid protein interacts with specific host factors to facilitate PIC nuclear import, while additional interactions of viral integrase, the enzyme responsible for viral DNA integration, with cellular nuclear proteins and nucleobases guide integration to specific chromosomal sites. HIV-1 integration favors transcriptionally active chromatin such as speckle-associated domains and disfavors heterochromatin including lamina-associated domains. In this review, we describe virus-host interactions that facilitate HIV-1 PIC nuclear import and integration site targeting, highlighting commonalities among factors that participate in both of these steps. We moreover discuss how the nuclear landscape influences HIV-1 integration site selection as well as the establishment of active versus latent virus infection.

CRISPR-Cas9 cytidine and adenosine base editing of splice-sites mediates highly-efficient disruption of proteins in primary cells

ABSTRACTBase editors allow for precise nucleotide editing without the need for genotoxic double-stranded breaks. Prior work has used base editors to knockout genes by introducing premature stop codons or by disrupting conserved splice-sites, but no direct comparison exists between these methods. Additionally, while base editor mediated disruption of splice sites has been used to shift the functional isoform pool, its utility for gene knockout requires further validation. To address these needs, we developed the program SpliceR (z.umn.edu/spliceR) to design cytidine-deaminase base editor (CBE) and adenosine-deaminase base editor (ABE) splice-site targeting guides. We compared the splice-site targeting and premature stop codon introduction in a knockout screen against the TCR-CD3 immune synapse in primary human T-cells. Our data suggests that 1) the CBE, BE4 is more reliable than the ABE, ABE7.10 for splice-site targeting knockout and 2) for both CBEs and ABEs, splice-donor targeting is the most reliable approach for base editing induced knockout.

Specialized DNA structures act as genomic beacons for integration by evolutionarily diverse retroviruses

ABSTRACTRetroviral integration site targeting is not random and plays a critical role in expression and long-term survival of the integrated provirus. To better understand the genomic environment surrounding retroviral integration sites, we performed an extensive comparative analysis of new and previously published integration site data from evolutionarily diverse retroviruses from seven genera, including different HIV-1 subtypes. We showed that evolutionarily divergent retroviruses exhibited distinct integration site profiles with strong preferences for non-canonical B-form DNA (non-B DNA). Whereas all lentiviruses and most retroviruses integrate within or near genes and non-B DNA, MMTV and ERV integration sites were highly enriched in heterochromatin and transcription-silencing non-B DNA features (e.g. G4, triplex and Z-DNA). Compared to in vitro-derived HIV-1 integration sites, in vivo-derived sites are significantly more enriched in transcriptionally silent regions of the genome and transcription-silencing non-B DNA features. Integration sites from individuals infected with HIV-1 subtype A, C or D viruses exhibited different preferences for non-B DNA and were more enriched in transcriptionally active regions of the genome compared to subtype B virus. In addition, we identified several integration site hotspots shared between different HIV-1 subtypes with specific non-B DNA sequence motifs present at these hotspots. Together, these data highlight important similarities and differences in retroviral integration site targeting and provides new insight into how retroviruses integrate into genomes for long-term survival.Graphical AbstractSchematic comparing integration site profiles from evolutionarily diverse retroviruses. Upper left, heatmaps showing the fold-enrichment (blue) and fold-depletion (red) of integration sites near non-B DNA features (lower left). Lower right, circa plot showing integration site hotspots shared between HIV-1 subtype A, B, C and D virus.