Identification Of Potential Drug Targets Research Articles

In Silico Subtractive Proteomics Approach for Identification of Potential Drug Targets in Staphylococcus saprophyticus.

Staphylococcus saprophyticus is a uropathogenic bacteria responsible for acute urinary tract infections (UTIs) mainly in young female patients. Patients suffering from urinary catheterization, pregnant patients, the elderly as well as those with nosocomial UTIs are at greater risk of the colonizing S. saprophyticus infection. The causative factors include benign prostatic hyperplasia, indwelling catheter, neurogenic bladder, pregnancy, and history of frequent UTIs. Recent findings have exhibited that S. saprophyticus is resistant to several antimicrobial agents. Moreover, there is a global concern regarding the increasing level of antimicrobial resistance, which leads to treatment failure and reduced effectiveness of broad-spectrum antimicrobials. Therefore, a novel approach is being utilized to combat resistant microbes since the past few years. Subtractive proteome analysis has been performed with the entire proteome of S. saprophyticus strain American Type Culture Collection (ATCC) 15305 using several bioinformatics servers and software. The proteins that were non-homologous to humans and bacteria were identified for metabolic pathway analysis. Only four cytoplasmic proteins were found possessing the potential of novel drug target candidates. The development of innovative therapeutic agents by targeting the inhibition of any essential proteins may disrupt the metabolic pathways specific to the pathogen, thus causing destruction as well as eradication of the pathogen from a particular host. The identified targets can facilitate in designing novel and potent drugs against S. saprophyticus strain ATCC 15305.

MAPPS: A Web-Based Tool for Metabolic Pathway Prediction and Network Analysis in the Postgenomic Era.

Comparative and evolutionary analyses of metabolic networks have a wide range of applications, ranging from research into metabolic evolution through to practical applications in drug development, synthetic biology, and biodegradation. We present MAPPS: Metabolic network Analysis and Pathway Prediction Server (https://mapps.lums.edu.pk), a web-based tool to study functions and evolution of metabolic networks using traditional and 'omics data sets. MAPPS provides diverse functionalities including an interactive interface, graphical visualization of results, pathway prediction and network comparison, identification of potential drug targets, in silico metabolic engineering, host-microbe interactions, and ancestral network building. Importantly, MAPPS also allows users to upload custom data, thus enabling metabolic analyses on draft and custom genomes, and has an 'omics pipeline to filter pathway results, making it relevant in today's postgenomic era.

Molecular Docking, Synthesis and anti-HIV-1 Protease Activity of Novel Chalcones.

Since the beginning of the HIV/AIDS epidemic, 75 million people have been infected with the HIV and about 32 million people have died of AIDS. Investigation of the molecular mechanisms critical to the HIV replication cycle led to the identification of potential drug targets for AIDS therapy. One of the most important discoveries is HIV-1 protease, an enzyme that plays an essential role in the replication cycle of HIV. The aim of the present study is to synthesize and investigate anti-HIV-1 protease activity of some chalcone derivatives with the hope of discovering new lead structure devoid drug resistance. 20 structurally similar chalcone derivatives were synthesized and their physico-chemical characterization was performed. Binding of chalcones to HIV-1 protease was investigated by fluorimetric assay. Molecular docking studies were conducted to understand the interactions. The obtained results revealed that all compounds showed anti-HIV-1 protease activity. Compound C1 showed the highest inhibitory activity with an IC50 value of 0.001 μM, which is comparable with commercial product Darunavir. It is difficult to provide general principles of inhibitor design. Structural properties of the compounds are not the only consideration; ease of chemical synthesis, low molecular weight, bioavailability, and stability are also of crucial importance. Compared to commercial products the main advantage of compound C1 is the ease of chemical synthesis and low molecular weight. Furthermore, compound C1 has a structure that is different to peptidomimetics, which could contribute to its stability and bioavailability.

A comparative proteomic approach using metabolic pathways for the identification of potential drug targets against Helicobacter pylori.

Helicobacter pylori is the most highlighted pathogen across the globe especially in developing countries. Severe gastric problems like ulcers, cancers are associated with H. pylori and its prevalence is widespread. Evolution in the genome and cross-resistance with different antibiotics are the major reason of its survival and pandemic resistance against current regimens. To prioritize potential drug target against H. pylori by comparing metabolic pathways of its available strains. We used various computational tools to extract metabolic sets of all available (61) strains of H. pylori and performed pan genomics and subtractive genomics analysis to prioritize potential drug target. Additionally, the protein interaction and detailed structure-based studies were performed for further characterization of protein. We found 41 strains showing similar set of metabolic pathways. However, 19 strains were found with unique set of metabolic pathways. The metabolic set of these 19 strains revealed 83 unique proteins and BLAST against human proteome further funneled them to 38 non-homologous proteins. The druggability and essentiality testing further converged our findings to a single unique protein as a potential drug target against H. pylori. We prioritized one protein-based drug target which upon subject to applied protocol was found as close homolog of the Saccharopine dehydrogenase. Our study has opened further avenues of research regarding the discovery of new drug targets against H. pylori.

Dynamical Mechanisms of Glutamate Receptor Gating and Sub-Conductance

Ionotropic glutamate receptors (iGluR) are responsible for initiation of excitatory signal transmission in the central nervous system (CNS). They play important roles in synaptic plasticity and long-term potentiation, while their dysfunction causes variety of diseases including neurodegenerative diseases and epilepsy, therefore they are potential targets for drug design. Glutamate receptors exhibit a distinctly modular multi-domain structure, in which truncated proteins containing only the ligand binding domain (LBD) and the transmembrane domains (TM) form the smallest functional glutamate gated channels. These homo-tetrameric AMPA subtype glutamate receptors gating and desensitization occurs on millisecond time-scales, and thus, these are the smallest and fastest working ligand-gated ion channels. In this work we report on extensive multy-microsecond molecular dynamics (MD) simulations of the AMPA receptor gating and regulation. The simulations were designed to allow the receptors to inter-convert between their various functional states, such as open and closed TM channel, and ligand-bound, active and desensitized LBD domain. We have used a variety of machine learning (ML) methods and systematic feature selection techniques to characterize which specific features of the LBD structure are responsible for transducing the signal for channel opening. We have observed multiple events of partial closures of the channel correlated with the LBD cleft closure degree from which we are able to draw conclusions on specific mechanism of allosteric coupling between the LBD domains monomeric and quarternary structures, as well as the ion channel state of the TM domains. Our study is the first one that was able to successfully design a simulation for opening and closing of the TM ion channel in response to the state of the LBD domain and characterize thus far elusive sub-conductance states.

Structural Diversity of Sense and Antisense RNA Hexanucleotide Repeats Associated with ALS and FTLD.

The hexanucleotide expansion GGGGCC located in C9orf72 gene represents the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia (FTLD). Since the discovery one of the non-exclusive mechanisms of expanded hexanucleotide G4C2 repeats involved in ALS and FTLD is RNA toxicity, which involves accumulation of pathological sense and antisense RNA transcripts. Formed RNA foci sequester RNA-binding proteins, causing their mislocalization and, thus, diminishing their biological function. Therefore, structures adopted by pathological RNA transcripts could have a key role in pathogenesis of ALS and FTLD. Utilizing NMR spectroscopy and complementary methods, we examined structures adopted by both guanine-rich sense and cytosine-rich antisense RNA oligonucleotides with four hexanucleotide repeats. While both oligonucleotides tend to form dimers and hairpins, the equilibrium of these structures differs with antisense oligonucleotide being more sensitive to changes in pH and sense oligonucleotide to temperature. In the presence of K+ ions, guanine-rich sense RNA oligonucleotide also adopts secondary structures called G-quadruplexes. Here, we also observed, for the first time, that antisense RNA oligonucleotide forms i-motifs under specific conditions. Moreover, simultaneous presence of sense and antisense RNA oligonucleotides promotes formation of heterodimer. Studied structural diversity of sense and antisense RNA transcripts not only further depicts the complex nature of neurodegenerative diseases but also reveals potential targets for drug design in treatment of ALS and FTLD.

Analysis of lncRNA, miRNA and mRNA-associated ceRNA networks and identification of potential drug targets for drug-resistant non-small cell lung cancer.

Background: Drug resistance to chemotherapeutic drugs or targeted medicines is an obstacle encountered in the treatment of non-small-cell lung cancer (NSCLC). However, the mechanisms of competing endogenous RNA (ceRNA) on the drug resistance in NSCLC are rarely reported. In this paper, the comprehensive expression profiles of lncRNAs and mRNAs in drug-resistant NSCLC cells were obtained by RNA sequencing.Methods: The dysregulated lncRNAs, miRNAs and mRNAs in drug-resistant NSCLC cell lines were identified by RNA-sequencing and bioinformatics methods.Results: A total of 39 dysregulated lncRNAs and 650 dysregulated mRNAs were identified between drug-resistant NSCLC cell lines and their parental cell lines. Additionally, 33 lncRNA-miRNA-mRNA pathways in the ceRNA network in drug-resistant NSCLC were constructed through bioinformatics methods and ceRNA regulatory rules. These comprised 12 dysregulated lncRNAs, five dysregulated miRNAs, and eight dysregulated mRNAs. In addition, lncRNA ATP2B1/miR-222-5p/TAB2 and lncRNA HUWE1/miR-222-5p/TAB2 were identified as potential ceRNA networks involved in drug resistance to NSCLC.Conclusions: The current study provides a promising therapeutic strategy against the lncRNA-miRNA-mRNA ceRNA regulatory network for NSCLC treatment and deepens our comprehension of the ceRNA regulatory mechanisms related to drug resistance to NSCLC.

IPSC-derived hepatocytes generated from NASH donors provide a valuable platform for disease modeling and drug discovery.

ABSTRACTNon-alcoholic fatty liver disease (NAFLD) affects 30–40% of adults and 10% of children in the US. About 20% of people with NAFLD develop non-alcoholic steatohepatitis (NASH), which may lead to cirrhosis and liver cancer, and is projected to be a leading cause of liver transplantation in the near future. Human induced pluripotent stem cells (iPSC) from NASH patients are useful for generating a large number of hepatocytes for NASH modeling applications and identification of potential drug targets. We developed a novel defined in vitro differentiation process to generate cryopreservable hepatocytes using an iPSC panel of NASH donors and apparently healthy normal (AHN) controls. iPSC-derived hepatocytes displayed stage specific phenotypic markers, hepatocyte morphology, with bile canaliculi. Importantly, both fresh and cryopreserved definitive endoderm and hepatoblasts successfully differentiated to pure and functional hepatocytes with increased CYP3A4 activity in response to rifampicin and lipid accumulation upon fatty acid (FA) treatment. End-stage hepatocytes integrated into three-dimensional (3D) liver organoids and demonstrated increased levels of albumin secretion compared to aggregates consisting of hepatocytes alone. End-stage hepatocytes derived from NASH donors demonstrated spontaneous lipidosis without FA supplementation, recapitulating a feature of NASH hepatocytes in vivo. Cryopreserved hepatocytes generated by this protocol across multiple donors will provide a critical cell source to facilitate the fundamental understanding of NAFLD/NASH biology and potential high throughput screening applications for preclinical evaluation of therapeutic targets.

Bioluminescent Probe for Monitoring Endogenous Fibroblast Activation Protein-Alpha.

Fibroblast activation protein-α (FAP), as a crucial member of cell surface glycoprotein, highly expresses in reactive fibroblasts of tumors and several fibrosis diseases. It is a potential target for drug design and also reported as a prodrug strategy to increase the therapeutic window of some anticancer agents. In this work, we developed the first bioluminogenic probe for FAP with a limit-of-detection of 0.254 ng/mL, which could be applied to evaluate the FAP inhibitors in vitro. The experiments of transgenic mice and tumor-bearing nude mice validated our probe 1 could reflect the endogenous FAP level in vivo. Furthermore, this probe was successfully used to reflect FAP up-regulation in the lung homogenates of the bleomycin-induced idiopathic pulmonary fibrosis mice.

Adenosine Receptors in Modulation of Central Nervous System Disorders.

The ubiquitous signaling nucleoside molecule, adenosine is found in different cells of the human body to provide its numerous pharmacological role. The associated actions of endogenous adenosine are largely dependent on conformational change of the widely expressed heterodimeric G-protein-coupled A1, A2A, A2B, and A3 adenosine receptors (ARs). These receptors are well conserved on the surface of specific cells, where potent neuromodulatory properties of this bioactive molecule reflected by its easy passage through the rigid blood-brainbarrier, to simultaneously act on the central nervous system (CNS). The minimal concentration of adenosine in body fluids (30-300 nM) is adequate to exert its neuromodulatory action in the CNS, whereas the modulatory effect of adenosine on ARs is the consequence of several neurodegenerative diseases. Modulatory action concerning the activation of such receptors in the CNS could be facilitated towards neuroprotective action against such CNS disorders. Our aim herein is to discuss briefly pathophysiological roles of adenosine on ARs in the modulation of different CNS disorders, which could be focused towards the identification of potential drug targets in recovering accompanying CNS disorders. Researches with active components with AR modulatory action have been extended and already reached to the bedside of the patients through clinical research in the improvement of CNS disorders. Therefore, this review consist of recent findings in literatures concerning the impact of ARs on diverse CNS disease pathways with the possible relevance to neurodegeneration.

Screening and Evaluation of Multi Drug Resistance Activity of Various 1,2,4-Triazoles Derivatives through in Vitro Anti tubercular

Isoniazid (Isonicotinic acid hydrazide, INH) has been significantly used to treat Mycobacterium tuberculosis. Introduction of drugs like INH, Rifmapicin, Pyrazinamide and Streptomycin resulted in rapid decline in TB cases worldwide. Several factors lead to the emergence of resistant strains of Mycobacterium tuberculosis. HIV infection also contributed to the escalating burden of tuberculosis. In the present examination 1,2,4-triazole subordinates were planned, incorporated and exposed to in vitro antitubercular screening against Mycobacterium tuberculosis H37Rv.Lipophilicity (log P) of the compounds were also determined to establish a correlation ship between physicochemical properties and antitubercular activity. Mtb CYP121 and CYP125 are considered to be potential targets for drug design. Binding study of azoles with these enzymes have also been reported. However, enough reports are not available on Mtb CYP-ligand binding requirements to improve the MIC of Azole based antitubercular agents. Hence we conducted the docking study of our synthesized triazoles against both Mtb CYP 121 and CYP125 to establish a correlationship between antitubercular activity and receptor binding interactions. In this paper we discuss about the molecular docking studies of the synthesized mercapto and benzthio 1,2,4-triazole compounds 13-18 with different enzyme target which we have employed.

Structural Analysis and Dynamic Processes of the Transmembrane Segment Inside Different Micellar Environments-Implications for the TM4 Fragment of the Bilitranslocase Protein.

The transmembrane (TM) proteins are gateways for molecular transport across the cell membrane that are often selected as potential targets for drug design. The bilitranslocase (BTL) protein facilitates the uptake of various anions, such as bilirubin, from the blood into the liver cells. As previously established, there are four hydrophobic transmembrane segments (TM1–TM4), which constitute the structure of the transmembrane channel of the BTL protein. In our previous studies, the 3D high-resolution structure of the TM2 and TM3 transmembrane fragments of the BTL in sodium dodecyl sulfate (SDS) micellar media were solved using Nuclear Magnetic Resonance (NMR) spectroscopy and molecular dynamics simulations (MD). The high-resolution 3D structure of the fourth transmembrane region (TM4) of the BTL was evaluated using NMR spectroscopy in two different micellar media, anionic SDS and zwitterionic DPC (dodecylphosphocholine). The presented experimental data revealed the existence of an -helical conformation in the central part of the TM4 in both micellar media. In the case of SDS surfactant, the -helical conformation is observed for the Pro258–Asn269 region. The use of the zwitterionic DPC micelle leads to the formation of an amphipathic -helix, which is characterized by the extension of the central -helix in the TM4 fragment to Phe257–Thr271. The complex character of the dynamic processes in the TM4 peptide within both surfactants was analyzed based on the relaxation data acquired on N and P isotopes. Contrary to previously published and present observations in the SDS micelle, the zwitterionic DPC environment leads to intensive low-frequency molecular dynamic processes in the TM4 fragment.

Potential Drug Target Identification of Legionella pneumophila by Subtractive Genome Analysis: An In Silico Approach

Though Legionella pneumophila is an opportunistic pathogen, recent reports about multi drug resistance in L. pneumophila is alarming. Annotated whole genome provides a pool of information which is applied for therapeutic drug targets identification in pathogenic bacteria. Subtractive genomic analysis is a pragmatic approach to screen the essential proteins present in pathogen but absent in host. Phylogenetically closely related L. pneumophila str. Philadelphia and L. pneumophila str. ATCC43209 protein profiles were analyzed to identify putative drug targets. Paralogous duplicate profiles were primarily discarded using CD-hit suit. Six hundred and ninety one L. pneumophila str. Philadelphia and 690 L. pneumophila str. ATCC43209 human homologous proteins were excluded using blstP. Among the human non-homologous proteins, the essential proteins for bacteria were separated using DEG tool. For both strains, one hundred and nineteen essential proteins were marked which participate in various metabolic pathways. Among them 11 unique proteins were found. Beside it, 15 and 16 exposed surface proteins were present in strain Philadelphia and ATCC43209 respectively. These unique and cell surface proteins can be utilized for effective drug and vaccine targets.
 Bangladesh J Microbiol, Volume 35 Number 2 December 2018, pp 102-107

H3.3K27M-induced chromatin changes drive ectopic replication through misregulation of the JNK pathway in C. elegans

Substitution of lysine 27 with methionine in histone H3.3 is a recently discovered driver mutation of pediatric high-grade gliomas. Mutant cells show decreased levels and altered distribution of H3K27 trimethylation (H3K27me3). How these chromatin changes are established genome-wide and lead to tumorigenesis remains unclear. Here we show that H3.3K27M-mediated alterations in H3K27me3 distribution result in ectopic DNA replication and cell cycle progression of germ cells in Caenorhabditis elegans. By genetically inducing changes in the H3.3 distribution, we demonstrate that both H3.3K27M and pre-existing H3K27me3 act locally and antagonistically on Polycomb Repressive Complex 2 (PRC2) in a concentration-dependent manner. The heterochromatin changes result in extensive gene misregulation, and genetic screening identified upregulation of JNK as an underlying cause of the germcell aberrations. Moreover, JNK inhibition suppresses the replicative fate in human tumor-derived H3.3K27M cells, thus establishing C. elegans as a powerful model for the identification of potential drug targets for treatment of H3.3K27M tumors.

In silico Identification of potential Drug Target and Analysis of Effective Single Nucleotide Polymorphisms for autism spectrum Disorder

Autism or Autism spectrum disorder is one of the most heritable neurophyschitric disorders; its underlying genetic architecture has largely eluded description. However the advances in genetics and genomics science and technology have the potential to revolutionize health care, so the number of reports investigating disease susceptibility based on the carriage of low-penetrance, high frequency SNPs has increased in recent years as SNPs are the most important and basic form of variation in the genome, and they are responsible for genetic effects that produce susceptibility to most autoimmune diseases. Here, the identification of novel target genes of ASD were retrieved and validated in subjected to Autdb, GeneCards, MalaCards, STRING and many others, which was resulted total 86 numbers of target genes for ASD. Thereafter the collection of associated Single Nucleotide Polymorphisms of the target genes became efficaciously carried out with the involvement of SIFT, PolyPhen2, PROVEAN and SNAP2, which results the most deleterious SNP with rsIDs rs35859650, rs34526199 and rs61752478 of selected target gene AMPD1 against ASD. The finding of this current study followed by GWAS not solely revealed the foremost necessary target genes for ASD, however conjointly it provides the associated single nucleotide polymorphisms can take one step nearer or forward to unravel the complicated structure of common deadly diseases especially mental disorders like Autism Spectrum Disorder.

Fatty Acid Signaling Mechanisms in Neural Cells: Fatty Acid Receptors.

Fatty acids (FAs) are typically associated with structural and metabolic roles, as they can be stored as triglycerides, degraded by β-oxidation or used in phospholipids’ synthesis, the main components of biological membranes. It has been shown that these lipids exhibit also regulatory functions in different cell types. FAs can serve as secondary messengers, as well as modulators of enzymatic activities and substrates for cytokines synthesis. More recently, it has been documented a direct activity of free FAs as ligands of membrane, cytosolic, and nuclear receptors, and cumulative evidence has emerged, demonstrating its participation in a wide range of physiological and pathological conditions. It has been long known that the central nervous system is enriched with poly-unsaturated FAs, such as arachidonic (C20:4ω-6) or docosohexaenoic (C22:6ω-3) acids. These lipids participate in the regulation of membrane fluidity, axonal growth, development, memory, and inflammatory response. Furthermore, a whole family of low molecular weight compounds derived from FAs has also gained special attention as the natural ligands for cannabinoid receptors or key cytokines involved in inflammation, largely expanding the role of FAs as precursors of signaling molecules. Nutritional deficiencies, and alterations in lipid metabolism and lipid signaling have been associated with developmental and cognitive problems, as well as with neurodegenerative diseases. The molecular mechanism behind these effects still remains elusive. But in the last two decades, different families of proteins have been characterized as receptors mediating FAs signaling. This review focuses on different receptors sensing and transducing free FAs signals in neural cells: (1) membrane receptors of the family of G Protein Coupled Receptors known as Free Fatty Acid Receptors (FFARs); (2) cytosolic transport Fatty Acid-Binding Proteins (FABPs); and (3) transcription factors Peroxisome Proliferator-Activated Receptors (PPARs). We discuss how these proteins modulate and mediate direct regulatory functions of free FAs in neural cells. Finally, we briefly discuss the advantages of evaluating them as potential targets for drug design in order to manipulate lipid signaling. A thorough characterization of lipid receptors of the nervous system could provide a framework for a better understanding of their roles in neurophysiology and, potentially, help for the development of novel drugs against aging and neurodegenerative processes.

Genome-wide association study of medication-use and associated disease in the UK Biobank

Genome-wide association studies (GWASs) of medication use may contribute to understanding of disease etiology, could generate new leads relevant for drug discovery and can be used to quantify future risk of medication taking. Here, we conduct GWASs of self-reported medication use from 23 medication categories in approximately 320,000 individuals from the UK Biobank. A total of 505 independent genetic loci that meet stringent criteria (P < 10−8/23) for statistical significance are identified. We investigate the implications of these GWAS findings in relation to biological mechanism, potential drug target identification and genetic risk stratification of disease. Amongst the medication-associated genes are 16 known therapeutic-effect target genes for medications from 9 categories. Two of the medication classes studied are for disorders that have not previously been subject to large GWAS (hypothyroidism and gastro-oesophageal reflux disease).

Structural Studies of GMP Reductase from Escherichia coli

Guanosine Monophosphate Reductase (GMPR) is an NADPH‐dependent enzyme that catalyzes the reductive deamination of guanosine (G) monophosphate nucleotides into their corresponding inosine (I) monophosphate derivatives. GMPR plays an important role in purine salvage, re‐utilization of free intracellular bases, and contributes overall to the intracellular balance of G and adenine (A) nucleotides. As with many other enzymes in the purine salvage or biosynthesis pathways, GMPR is widespread in nature across eukaryotic and prokaryotic taxa. Given the interest in purine metabolism as a potential target for drug design, GMPR has been characterized both structurally and kinetically in different studies using GMPR derived from several species. We report progress towards solving the crystal structure of GMPR from Escherichia coli in order to extend the insight from existing studies of the E. coli GMPR kinetic and thermodynamic characteristics. Additionally, lessons learned from integrating related research activities into an undergraduate laboratory course will be presented in order to promote adoption of similar course‐based research practices at other primarily‐undergraduate institutions.Support or Funding InformationIthaca College Department of ChemistryThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

MRNA expression of chemosensitivity genes as a predictor of response to neoadjuvant chemotherapy in non-small cell lung cancer

The objective response rate to neoadjuvant chemotherapy (NAC) in patients with non-small cell lung cancer (NSCLC) is approximately 16 %. Therefore, the search for new predictive factors and markers that could precisely predict the NAC response and be relevant to the choice of the adequate treatment policy is of utmost importance. the purpose of the study was to analyze the relationship between the expression levels of chemosensitivity genes and NAC response in patients with non-small cell lung cancer. material and methods. The study included 30 patients with stage IIAIIIB NSCLC. Total RNA was isolated from normal and tumor lung tissue samples prior to NAC. The expression level of chemosensitivity genes, such as BRCA1, RRM1, ERCC1, TOP1, TOP2a, TUBB3, TYMS, and GSTP1 was evaluated by real-time reverse transcriptase quantitative PCR (RT-qPCR). results. A significant difference in the expression levels of the studied genes between patients with different NAC response was found. The objective response to therapy was observed in 67 % (10/15) of patients having no ERCC1 expression compared to those with ERCC1 expression (p&lt;0.05). Low (less than 0.2) and high (more than 1.2) BRCA1 expression levels were associated with low rates of NAC response compared with patients whose expression level was between the lower and upper quartiles. Statistically significant differences were shown for the GSTP1 and RRM1 genes. conclusion. A comprehensive assessment of the expression of chemosensitivity genes is important not only in terms of understanding heterogeneity and complexity of the molecular biology of NSCLC, but also for more accurate prediction of response to NAC and identification of potential drug targets.

Mutations in the dimer interfaces of the dengue virus capsid protein affect structural stability and impair RNA-capsid interaction

The dengue virus 2 capsid protein (DENV2C) plays a primary structural role in the protection of the viral genome and is crucial for nucleocapsid assembly. In this study, we generated single mutants of DENV2C at L50 and L54 residues of the α2 helix, which was shown to interfere with the integration of the capsid into lipid droplets, and at residues L81 and I88 located in the α4 helix, which was shown to affect viral assembly. We demonstrated that the oligomeric states of DENV2C and its mutants exist primarily in the dimeric state in solution. All single-point mutations introduced in DENV2C promoted reduction in protein stability, an effect that was more pronounced for the L81N and I88N mutants, but not protein unfolding. All the single-point mutations affected the ability of DEN2C to interact with RNA. We concluded that mutations in the α2-α2′ and α4-α4′ dimer interfaces of DENV2C affect the structural stability of the protein and impair RNA-capsid interaction. These effects were more pronounced for mutations at the L81 and I88 residues in the α4 helix. These results indicate the importance of the α4-α4′ dimer interface, which could be studied as a potential target for drug design in the future.