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Continued advancements in CRISPR-Cas systems have accelerated genome research. Use of CRISPR-Cas in cancer research has been of great interest that is resulting in development of orthogonal methods for drug target validations and discovery of new therapeutic targets through genome-wide screens of cancer cells. CRISPR-based screens have also revealed several new cancer drivers through alterations in tumor suppressor genes (TSGs) and oncogenes inducing resistance to targeted therapies via activation of alternate signaling pathways. Given such dynamic status of cancer, we review the application of CRISPR-Cas in non-small cell lung cancer (NSCLC) for development of mutant models, drug screening, target validation, novel target discoveries, and other emerging potential applications. In addition, CRISPR-based approach for development of novel anticancer combination therapies is also discussed in this review.

Supplementary Materials: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/ijms23063045/s1. Supporting information consisting of a blot testing cross reactivity of anti-C1q against IAV lysate, C1q and recombinant globular head cytotoxicity panel and the cytokine panel. Copyright: © 2022 by the authors. The Influenza A virus (IAV) is a severe respiratory pathogen. C1q is the first subcomponent of the complement system’s classical pathway. C1q is composed of 18 polypeptide chains. Each of these chains contains a collagen-like region located at the N terminus, and a C-terminal globular head region organized as a heterotrimeric structure (ghA, ghB and ghC). This study was aimed at investigating the complement activation-independent modulation by C1q and its individual recombinant globular heads against IAV infection. The interaction of C1q and its recombinant globular heads with IAV and its purified glycoproteins was examined using direct ELISA and far-Western blotting analysis. The effect of the complement proteins on IAV replication kinetics and immune modulation was assessed by qPCR. The IAV entry inhibitory properties of C1q and its recombinant globular heads were confirmed using cell binding and luciferase reporter assays. C1q bound IAV virions via HA, NA and M1 IAV proteins, and suppressed replication in H1N1, while promoting replication in H3N2-infected A549 cells. C1q treatment further triggered an anti-inflammatory response in H1N1 and pro-inflammatory response in H3N2-infected cells as evident from differential expression of TNF-α, NF-κB, IFN-α, IFN-β, IL-6, IL-12 and RANTES. Furthermore, C1q treatment was found to reduce luciferase reporter activity of MDCK cells transfected with H1N1 pseudotyped lentiviral particles, indicative of an entry inhibitory role of C1q against infectivity of IAV. These data appear to demonstrate the complement-independent subtype specific modulation of IAV infection by locally produced C1q. Funding: This research received no specific grant from any funding agency. Funding: This research received no specific grant from any funding agency.

Influenza A virus (IAV) is a contagious respiratory infection causing pathogen responsible for high morbidity and mortality rates across the planet. The human immune system contains a wide range of soluble activators, membrane-bound receptors, and regulators to eliminate IAVs. Despite these various immune mechanisms that neutralize IAVs or restrict their replication, IAVs have developed distinct strategies to evade host immunity and establish a successful infection. Given the higher and continuous rate of mutations in IAVs, decades of research have focused on understanding the host's immune mechanisms against IAVs, and the evasion strategies employed by the virus to overcome the host immune system. Future IAV pandemics or epidemics remain inevitable, and a greater understanding of the host-pathogen interaction involved is required to develop universal vaccines and treatments against IAV. Here, we review how the host immune system responds to IAV infection as well as the strategies employed by the IAV to evade host immune surveillance. Furthermore, this review also focuses on the treatments and vaccines that have been developed to counter IAV infection.

Delivery of small-interfering RNA (siRNA) has been of great interest in the past decade for effective gene silencing. To overcome synthetic and regulatory challenges posed by nanoparticle-mediated siRNA delivery, antibody–siRNA conjugate (ARC) platform is emerging as a potential siRNA delivery system suitable for clinical translation. Herein, we have developed a delivery technology based on the ARC platform for stable delivery of siRNA called as Gelatin-Antibody Delivery System (GADS). In GADS, positively charged gelatin acts as a linker between antibody–siRNA and enables the endosomal escape of siRNA for gene silencing postcellular internalization. For proof of concept, we synthesized a scalable GADS conjugate comprising of Cetuximab (CTB), cationized gelatin (cGel) and NSCLC KRASG12C-specific siRNA. CTB was chemically conjugated to cGel through an amide link to form the CTB–cGel complex. Thereafter, siRNA was chemically conjugated to the cGel moiety of the complex through the thioether link to form CTB–cGel–siRNA conjugate. RP-HPLC analysis was used to monitor the reaction while gel retardation assay was used to determine siRNA loading capacity. SPR analysis showed the preservation of ligand binding affinity of antibody conjugates with KD of ∼0.3 nM. Furthermore, cellular internalization study using florescent microscopy revealed receptor-mediated endocytosis. The conjugate targeted EGFR receptor of KRAS mutant NSCLC to specifically knockdown G12C mutation. The oncogene knockdown sensitized the cells toward small molecule inhibitor—Gefitinib causing ∼70% loss in cell viability. Western blot analysis revealed significant downregulation for various RAS downstream proteins postoncogene knockdown. Comparison of the efficiency of GADS vis-à-vis positive siRNA control and CRISPR–Cas9-based knockout of KRAS Exon 2 in the NCI-H23 NSCLC cell line suggests GADS as a potential technology for clinical translation of gene therapy.

Continued advancements in CRISPR-Cas systems have accelerated genome research. Use of CRISPR-Cas in cancer research has been of great interest that is resulting in development of orthogonal methods for drug target validations and discovery of new therapeutic targets through genome-wide screens of cancer cells. CRISPR-based screens have also revealed several new cancer drivers through alterations in tumor suppressor genes (TSGs) and oncogenes inducing resistance to targeted therapies via activation of alternate signaling pathways. Given such dynamic status of cancer, we review the application of CRISPR-Cas in non-small cell lung cancer (NSCLC) for development of mutant models, drug screening, target validation, novel target discoveries, and other emerging potential applications. In addition, CRISPR-based approach for development of novel anticancer combination therapies is also discussed in this review.

Alopecia or hair loss is a worldwide unisex dermatological problem which affects aesthetic lifestyle qualities in humans. In recent years, drug discovery for hair loss has gained significant pharmaceutical research attention. Synthetic drugs such as minoxidil, oral finasteride, anthralin cream, and ketoconazole-based antifungal shampoos are some of the commercially available product formulations for hair loss treatment. As these products are mostly chemically derived, their long-term exposure to the skin could result in various side effects and skin disorders. As traditional medicine relies on herbs to treat alopecia, in recent times, different species of herbs are being extracted to generate functional bioactive chemicals as active ingredients to treat hair loss. These biologically derived phytochemicals may offer improved long-term biocompatibility with the skin. This chapter presents an overview of various phytochemicals with antialopecia properties and discusses their modes of action. Additionally, the efficiency of flavonoids, which is a major phytochemical constituent of several herbs and a potential 5α-reductase enzyme inhibitor, as a potential drug for alopecia treatment is also discussed.

Androgenetic alopecia (AGA) is a progressive dermatological disorder of frontal and vertex scalp hair loss leading to baldness in men. This study aimed to identify candidate genes involved in AGA through an in silico search strategy. The gene expression profile GS36169, which contains microarray gene expression data from bald frontal and haired occipital scalps of five men with AGA, was downloaded from the Gene Expression Omnibus (GEO) database. The differential gene expression analysis for all five subjects was carried out separately by PUMA package in R and identified 32 differentially expressed genes (DEGs) common to all five subjects. Gene ontology (GO) biological process and pathway- enrichment analyses of the DEGs were conducted separately for the up-regulated and down-regulated genes. ReactomeFIViz app was utilized to construct the protein functional interaction network for the DEGs. Through GO biological process and pathway analysis on the clusters of the Reactome FI network, we found that the down-regulated DEGs participate in Wnt signaling, TGF-beta signaling, and up-regulated DEGs participate in oxidative-stress- related pathways.

Background: Reported literature on invasive group A streptococcal isolates in India is very scanty. This study was undertaken to determine the molecular heterogeneity of such isolates as seen in a tertiary care center. Materials and Methods: Thirty two blood culture isolates and 18 from other sterile body fluids were characterized by emm gene sequencing and multilocus sequence typing. Results: Forty two emm types were identified including 25 from 32 blood isolates and 17 from 18 other body fluid isolates. Types 110, 74, 63, 85, 102, 105, 124 and st854.1 were common to both groups and accounted for 40% of the isolates. Two types namely, stKNB6 and stKNB9 were newly identified types. MLST identified forty eight sequence types (MLST - ST) of which 31 were from 32 blood isolates and 17 from 18 body fluid isolates; thirty three of them were hitherto unrecognized at the time of identification. Two blood isolates of emm 85 had the same MLST - ST 484 while three blood isolates of emm 110 had three different STs namely, ST 493, 494 and 497. Two types, ST 493 and ST497 had single locus variation while ST 497 had a double locus variation. Conclusions: Our study shows that subtle allelic variations in the house keeping genes results in the development of new strains in a given emm type and contribute significantly to the existing high diversity of strains circulating in the community.

Androgenetic alopecia (AGA) is a dermatological disorder of scalp hair loss characterized by a progressive miniaturization of hair follicles with shortened anagen phase leading to a decreased number of hairs on the scalp. It is a complex polygenic trait prevailing around two-thirds of the male population. Elevated expressions of 5α-dihydrotestosterone and androgen receptor are the causal factors for AGA. This review describes recent studies on the role of androgens and androgen receptor (AR) transactivation activity in modulating the Wnt/β-catenin signaling in the dermal papilla cells of the balding scalp in androgenetic alopecia. Here, we analyse the androgen-induced dermal papilla secreted factors on stimulating catagen entry in hair follicles and the molecular cross-talk between AR and Wnt/β-catenin signaling with a brief mention on alternative treatment strategy targeting Wnt/β-catenin signaling for promoting hair growth.

Food crisis is a major issue in underdeveloped and overpopulated countries. Due to this crisis, many people are prone to get affected by various health disorders and nutrition relation problems. The protein sources for vegetarians are very limited. Due to globalization, there is a change of lifestyles, and there is an increase in consumption of junk food. Consequently, there is a rise in the number of obese people. Protein supplements are expensive and not easily affordable for everyone. Hence, there arises a need for a protein source, which is consumed by everyone and has high nutritive value too. With the ever-increasing rise in pollution, the quality of food and vegetation is falling. Maybe due to globalization, the food choices are increasing, but the contamination and adulteration are rising too. Purity has become a major concern for people at present. To combat these problems, single cell protein from microorganism can be used as an optional nutritional source. This article talks about different microorganisms sources of single cell protein.

Benzene is a very common air pollutant and is known to be a carcinogen. It causes cancer in people who are exposed to benzene vapors over a very long period of time. Cancer is one of the leading causes of death, causing millions of death every year. Zebra fish is a very promising experimental model for drug testing and cancer studies. In this experiment benzene was used as a carcinogen in zebra fishes to check if it induces cancer in the fishes. These cancer induced fishes may be used for drug discovery and testing for cancer therapy. Different concentrations of benzene were injected into the zebra fishes, and the fishes were monitored over a period of 14 days. The fishes were observed for presence of any cancerous growth, and their weights were checked in order to determine any increase in mass. The results were inconclusive as no tumor growth was observed. This could be due to the short time over which they were monitored or the concentration of benzene being used may be low. May be if the time period, the period of exposure and the concentration of the benzene were increased, some significant results might be observed.

Tuberculosis, a transmittable killer disease caused by Mycobacterium tuberculosis whose pathogenesis is poorly understood. The treatment available for the anti-mycobacterial activity has poor observance because of the uprising of the drug resistance strains resulted by the prolonged treatment giving the chance to the microbe to develop resistance. Hence, there is an urgent need to identify novel compounds from marine sources which are having antimicrobial activity. GABA-AT is an important enzyme involved in GABA shunt pathway of TCA cycle which has a role in virulence of Mycobacterium tuberculosis as it is a PLP dependent enzyme. In this present study, the 3D structure of GABA-AT was generated by using SWISSMODEL and validated using various methods. The model generated was validated and secondary structural analysis was predicted using YASPIN. Thereafter, the active site was predicted and further used the residues for docking. The acquired model was docked with the known inhibitors of GABA-AT and marine compounds having anti-tuberculosis activity. The comparative analysis of protein ligand interaction between known inhibitor and marine compounds were studied. Among them, Heteronemin has the best binding energy (-9.19Kcal/mol) which indicates that the marine compound can be used for inhibiting the activity of GABA-AT in Mycobacterium tuberculosis and can be proposed as valuable drug candidate.

This neglected pathogen causes a large portion of these infections., Vellore, a region in southern India, has a high incidence of severe human infections with β-hemolytic group C and G streptococci (GCGS). To determine the causative species in these infections, we conducted 16S rRNA gene sequencing: Streptococcus dysgalactiae subsp. equisimilis (81%) and S. anginosus (19%) were the causative organisms in the 2-year study period (2006–2007). We used PCR to detect the virulence-related emm gene; results showed that it was restricted to S. dysgalactieae subsp. equisimilis isolates of 99.2% tested positive. Due to a novel marker, S. anginosus and S. constellatus can be quickly and accurately distinguished from other members of the genus. The notable contribution of the anginosus group to human infections suggests that this group of obligate pathogens deserves more attention in healthcare and research.

To evaluate the use of antibody detection kits in the diagnosis of pulmonary tuberculosis in an endemic area, serum samples from cases (sputum smear positive for AFB) and controls (healthy young adults) were collected and tested using five different kits. Sensitivity, specificity and predictive values were calculated using smear positivity as gold standard. Sensitivity of tests varied from 46% to 68% and the specificity from 68% to 100%. None of the kits evaluated can be used as a single screening test for tuberculosis. However kits with good specificity may be used in conjunction with conventional methods for diagnosis.