COVID-19 influenced gut dysbiosis, post-acute sequelae, immune regulation, and therapeutic regimens.

The novel coronavirus disease 2019 (COVID-19) pandemic outbreak caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has garnered unprecedented global attention. It caused over 2.47 million deaths through various syndromes such as acute respiratory distress, hypercoagulability, and multiple organ failure. The viral invasion proceeds through the ACE2 receptor, expressed in multiple cell types, and in some patients caused serious damage to tissues, organs, immune cells, and the microbes that colonize the gastrointestinal tract (GIT). Some patients who survived the SARS-CoV-2 infection have developed months of persistent long-COVID-19 symptoms or post-acute sequelae of COVID-19 (PASC). Diagnosis of these patients has revealed multiple biological effects, none of which are mutually exclusive. However, the severity of COVID-19 also depends on numerous comorbidities such as obesity, age, diabetes, and hypertension and care must be taken with respect to other multiple morbidities, such as host immunity. Gut microbiota in relation to SARS-CoV-2 immunopathology is considered to evolve COVID-19 progression via mechanisms of biochemical metabolism, exacerbation of inflammation, intestinal mucosal secretion, cytokine storm, and immunity regulation. Therefore, modulation of gut microbiome equilibrium through food supplements and probiotics remains a hot topic of current research and debate. In this review, we discuss the biological complications of the physio-pathological effects of COVID-19 infection, GIT immune response, and therapeutic pharmacological strategies. We also summarize the therapeutic targets of probiotics, their limitations, and the efficacy of preclinical and clinical drugs to effectively inhibit the spread of SARS-CoV-2.

Multidrug-resistant Bacillus species isolated from hospital soil environment is controlled by nanobiotics incorporated nanoformulation.

Multidrug resistance is a serious hazard to the environment, it claims a large number of lives every year. Lack of drug options and easy transmission of these organisms remain the biggest threat in treating the relative infections whose causative systems have evolved and become stronger in due course of time. Hospitals serve as one of the largest breeding grounds for harbouring these organisms. This study aims to isolate and characterize multidrug-resistant microorganisms from soil samples collected from hospital waste dumping premises. Polyherbal nanoformulation was synthesized from ethno-medicinal source Triphala (three berries) and characterized using various physico-chemical characterization techniques. The antibacterial efficacy of the polyherbal nanoformulation was evaluated by employing various assays to determine MIC, MBC, and biofilm inhibition potential in isolated strains. Bacterial colonies were isolated and the DNA was sequenced. The isolated organisms were identified as Bacillus cereus, Bacillus licheniformis, and Bacillus pumilus and they were subjected to antibiotic susceptibility by using various antibiotics. It was found that all the microorganisms were multidrug-resistant and possessed resistance to various classes of antibiotics. The various antibacterial assays showed that the polyherbal nanoformulation was highly effective in controlling growth and biofilm formation even at lower concentrations when compared with commercial antibiotics. The novelty of this research work lies in combining the beneficial effects of silver and polyherbal drugs into a single Polyherbal nanoformulation. This is the first novel report to utilize polyherbal nanoformulation to control the multidrug-resistant microorganisms thriving in hospital waste dumping sites. Hence, this nanobiotics incorporated polyherbal nanoformulation can be developed into a commercial product to treat the hospital waste material before dumping it into the environment.

Evaluation of Time-Dependent Corrosion Inhibition Rate for f-MWCNT-BCP Composite Coatings on 316L Stainless Steel in Simulated Body Fluid for Orthopedic Implantation.

A well-developed-multiwall carbon nanotube (f-MWCNT)/biphasic calcium phosphate (BCP) composites were synthesized using ultrasonication method for orthopedic implantation applications. The formation of composites and its phase was confirmed by using X-ray diffraction. The presence of various functional groups was identified by using Fourier transform infra-red (FT-IR) spectroscopy. The presence of f-MWCNT was confirmed by Raman spectroscopy. High-resolution transmission electron microscopy (HR-TEM) analysis revealed that BCP units were bound by the surface of f-MWCNTs. The synthesized composites were coated on medical grade 316L stainless steel substrates using electro deposition technique. To determine its corrosion resistance characteristics, the developed substrates were exposed to a simulated bodily fluid (SBF) solution for 0, 4, and 7 days. These results strongly suggest that the coated composites can be utilized for bone tissue repair.

Antibacterial, antibiofilm, and antivirulence effects of nanoparticles synthesized from Colletotrichum gloeosporioides in pathogenic E.coli.

Antimicrobial resistance is a global problem and antibiotics are becoming ineffective due to the resistance developed by bacteria. In this current research, silver nanoparticles were synthesized from aqueous extract of endophytic fungi Colletotrichum gloeosporioides (CgAgNPs) and characterized by various methods. CgAgNPs efficacy was analyzed by performing antimicrobial assays in Escherichia coli ATCC 25922 and antibiotic resistant pathogenic strains. Upon treatment with CgAgNPs biofilm formation was reduced in all E.coli strains. In vitro cytotoxicity assays revealed that CgAgNPs were able to increase the membrane permeability and induced leakage of sugars and proteins. CgAgNPs induced oxidative stress in E. coli strains led to lipid peroxidation and release of malonaldehyde. The CgAgNPs were able to modulate the anti-oxidant system of cells hence there was a reduction in Glutathione reductase, Catalase and Superoxide dismutase enzymes activities. Analysis of expression of gene encoding CTX-M-15 showed the down regulation upon treatment with ampicillin and CgAgNPs. Overall, the results suggest that CgAgNPs control growth, biofilm formation in E. coli through induction of oxidative stress, interference with antioxidant enzymes, cell content leakage and finally downregulating the virulence gene by interfering with transcription and translation in E. coli. In future, CgAgNPs can be incorporated in formulations to break antibiotic resistance in antibiotic resistant pathogenic E. coli.

Therapeutic potential of dietary phytochemicals from Drynaria quercifolia to modulate gut microbiome: an in silico approach.

Drynaria quercifolia is one of the pioneer medicinal plants which exert many beneficial effects on humans. Fatty acids are hydrophobic ligands that act as membrane substrates, metabolic signalling molecules, and metabolic energy sources. It could enhance the mucus production in the intestine which maintain mucosal homeostasis. The inflammatory responses were also regulated by 5-HT receptors. Serotonin 2 A receptors are G-protein coupled receptors targeted by various types of ligands viz. antidepressants, antipsychotics and anti-migraine drugs. The interaction of mucin protein and fatty acid could increase the antimicrobial activity and anti-inflammatory activity of gut microbiome. In this study, dietary phytochemicals were extracted from D. quercifolia and characterized using GC-MS analysis. The result shows presence of 38 various compounds viz. decanoic acid, indole, and dodecanoic acid and etc., Among the all, dodecanoic acid showed good drug likeness and pharmaceutical properties. Target proteins viz. 3IFB (Intestinal fatty acid binding protein) and 7WC4 (5-hydroxytryptamine receptor 2 A) were docked with dodecanoic acid using Auto dock software. The fatty acids produced in the gut could interact with 3IFB and 7WC4 proteins to maintain intestinal integrity and improved gut-brain function respectively. Dodecanoic acid exhibits the highest binding energy with mutated 3IFB (-6.01) than native whereas 7WC4 native protein showed (-7.3 kal/mol) the highest affinity than mutated protein. Structural changes were predicted by using SOPMA and ProSA. Evaluation results indicate that 3IFB was having more stability, good quality, and enhanced affinity than 7WC4. Further, MD simulations were done for 3IFB to check the stability of protein-ligand complex using WebGro platform. The model was investigated by root mean square deviation and fluctuations. Therefore, dodecanoic acids have been considered as a potential agonists and offer opportunities for developing innovative medications for gastrointestinal diseases.Communicated by Ramaswamy H. Sarma.

An In Silico Investigation of Pharmacological Modulators and Inflammasomes in Glioblastoma Multiforme.

For the past decades, inflammatory signals have been considered a possible key for pharmacological interventions. There are several compounds and/or molecules that have been known as most promising medication against inflammation and its mediated chronic disorders. Inflammasomes could be recognized as a trigger by detrimental stimuli as pathogenic attack and endogenous signals mediated injury inside the cells. In addition, there has been an inflammatory key mechanism involved in cancers including glioblastoma multiforme (GBM). GBM has been considered the foremost aggressive primary brain tumors in adult stage. There is a scattered beam of light on both cellular and molecular links in inflammation and GBM. However, the immune response of GBM has been characterized extensively by macrophages and lymphocytes related to tumors, and some recent investigations have pinpointed the focus of inflammasomes on the progression of GBM. Nevertheless, risk factors linked with GBM are still debatable. In our study, the most considerable compounds and their bonded and/or targeted proteins have depicted the most promising highlights under in silico condition. Our in silico investigations have revealed a powerful pharmacological agents/compound against inflammasome-mediated GBM.

State-of-the-art review on the ecotoxicology, health hazards, and economic loss of the impact of microcystins and their ultrastructural cellular changes.

Cyanobacteria are ubiquitously globally present in both freshwater and marine environments. Ample reports have been documented by researchers worldwide for pros and cons of cyanobacterial toxins. The implications of cyanobacterial toxin on health have received much attention in recent decades. Microcystins (MCs) represent the unique class of toxic metabolites produced by cyanobacteria. Although the beneficial aspects of cyanobacterial are numerous, the deleterious effect of MCs overlooked. Several studies on MCs evidently reported that MCs exhibit a plethora of harmful effect on animals, plants, and cell lines. Accordingly, numerous histopathological studies have also found that MCs cause detrimental effects to cells by damaging cellular organelles, including nuclear envelope, Golgi apparatus, endoplasmic reticulum, mitochondria, plastids, flagellum, pilus membrane structures and integrity, vesicle structures, and autolysosomes and autophagosomes. Such ultrastructural cellular damages holistically influence the morphological, biochemical, physiological, and genetic status of the host. Indeed, MCs have also been found to cause the deleterious effect to different animals and plants. Such deleterious effects of MCs have greater impact on agriculture, public health which in turn influences ecotoxicology and economic consequences. The impairments correspond to oxidative stress, organ failure, carcinogenesis, aquaculture loss, with an emphasis for blooms and respective bioaccumulation prospects. The preservation of mortality among life forms is addressed in a critical cellular perspective for multitude benefits. The comprehensive cellular assessment could provide opportunity to develop strategy for therapeutic implications.

Screening of Anti-carcinogenic Properties of Phytocompounds from Allium ascalonicum for Treating Breast Cancer Through In Silico and In Vitro Approaches.

Plants, rich in phytocompounds, have been in usage since time immemorial for treating various diseases, namely, cancer. One such plant species, Allium ascalonicum (Shallot) belonging to Amaryllidaceae family is being studied here for its anti-carcinogenic properties against breast cancer. GC-MS characterization of A. ascalonicum exhibited 48 phytocompounds containing five peak phytocompounds and 13 phytocompounds with anti-carcinogenic properties. These 13 anti-carcinogenic phytocompounds were docked with three hormonal receptors involved in breast cancer malignancy, namely, ERα, PR, and human EGFR with tamoxifen as standard for in silico analysis. The results exhibited three phytocompounds that had better binding scores compared to that of the standard drug, tamoxifen. Lyophilized powder of aqueous A. ascalonicum extract, also referred as ASE, was used for in vitro approaches. Antioxidant study using DPPH assay revealed that the highest percentage of FRSA in ASE, nearly 51%, was observed at 50 µg/ml concentration. Cytotoxicity study on MCF-7 cell line using MTT assay demonstrated IC50 value at 1400 µg/ml and anti-proliferative study using Trypan blue assay for the determination of percentage viability of MCF-7 cells at IC50 concentration was observed to be 49%. Anti-mitotic activity using Vigna radiata seed germination assay revealed clear morphological differences in a dose-dependent manner between the seeds grown at various concentrations of ASE with nearly 56.5% growth inhibition observed at 1500 µg/ml concentration. Hence, this research work proves that Allium ascalonicum has very good anti-carcinogenic properties and this can be confirmed further through in vivo animal model studies and it can also be formulated as a promising drug to treat breast cancer. GC-MS characterization of Allium ascalonicum demonstrated the presence of five peak compounds and thirteen anti-carcinogenic compounds. The thirteen anti-carcinogenic compounds were docked with three target proteins (in silico analysis) involved in breast cancer malignancy and identified the presence of three potential phytocompounds that can be used for treating breast cancer. In vitro approaches also confirmed the presence of anti-carcinogenic properties such as antioxidative potential, cytotoxic, anti-proliferative, and anti-mitotic effects. Hence, Allium ascalonicum can be taken further to in vivo studies so that it can be formulated to treat breast cancer.

Promising Action of Cannabinoids on ER Stress-Mediated Neurodegeneration: An In Silico Investigation.

Neurodegeneration has been recognized as a clinical episode characterized by neuronal death, including dementia, cognitive impairment and movement disorder. Most of the neurodegenerative deficits, via clinical symptoms, includes common pathogenic features as protein misfolding and aggregation. Therefore, the focus highlights the cellular organelle endoplasmic reticulum (ER) critically linked with the quality control and protein homeostasis. Unfolded protein response (UPR) or ER stress have also been considered as hallmarks for neurodegenerative disorders. It has been implicated that the levels of endocannabinoids (ECB) could rise at the platform of neurodegeneration. In addition, phytocannabinoids (PCB) including cannabidiol (CBD) could also initiate the IRE1, PERK, XBP-1, and ATF6, pathways that could lead to the degradation of the misfolded proteins and termination of protein translation. Thus, our aim was to determine if cannabinoids bind to these ER arm proteins involved in UPR by molecular docking and therefore determine its drug resemblance through ADME analysis. In our study, three cannabinoid receptors (CB1, CB2, and CB3) were considered to demonstrate their neuroprotective actions. The chosen ligands were screened as PCB (Δ9-tetrahydrocannabinol or THC), CBD, and two ECB, anandamide (AEA) and 2-arachidonoylglycerol (2-AG). The current findings have advocated that the cannabinoids and their molecular targets have shown considerable binding and their ADME properties also reveals that they possess moderate drug-like properties making it as a valuable option for the treatment and management of neurodegenerative diseases.

Engineered green nanoparticles interact with Nigrospora oryzae isolated from infected leaves of Arachis hypogaea.

Oilseed crop diseases are a major concern around the world, particularly in India. The synthetic fungicides not only kill the pathogen, but they also harm the host plant and beneficial microbes and on continuous usage, they decrease the soil fertility. To overcome this problem, green nanotechnology has been a greater alternative with promising benefits. The green synthesized nanoparticles from the extract of various plant parts are an effective remedy for killing the pathogens without affecting the host plants and the environment. Hence, in our study silver nanoparticles were synthesized from Fennel seed (Foeniculum vulgare) extract. The synthesis of nanoparticles was confirmed using UV-vis, Fourier transform infrared, dynamic light scattering, zeta potential, and scanning electron microscopic analysis. The in vitro antifungal study was carried out and revealed that the nanoparticles had high efficacy against the isolated phytopathogen Nigrospora oryzae which causes tikka disease in Arachis hypogaea plants. Hence, F. vulgare seed nanoparticles can be used as an effective alternative to synthetic fungicides without causing any deleterious effect on soil microflora or the environment.

Cellular and Molecular Machinery of Neuropathic Pain: an Emerging Insight.

Purpose of Review: Neuropathic pain (NP) has been ubiquitously characterized by lesion and its linked somatosensory system either the central nervous system (CNS) or peripheral nervous system (PNS) This PNS episode is the most prevalent site of NP origin and is found to be associated with afferent nerve fibers carrying pain signals from injured/trauma site to the CNS including the brain. Several kinds of pharmacotherapeutic drugs shuch as analgesics, anti-convulsants, and anti-depressants are being employed for the its possible interventions. The NP has been a great interest to follow different pathophysiological mechanisms which are often considered to correlate with the metabolic pathways and its mediated disease. There is paucity of knowledge to make such mechanism via NP. Recent Finding: Most notably, recent pandemic outbreak of COVID-19 has also been reported in chronic pain mediated diabetes, inflammatory disorders, and cancers. There is an increasing incidence of NP and its complex mechanism has now led to identify the possible investigations of responsible genes and proteins via bioinformatics tools. The analysis might be more instrumental as collecting the genes from pain genetic database, analyzing the variants through differential gene expression (DEG) and constructing the protein-protein interaction (PPI) networks and thereby determining their upregulating and downregulating pathways. Summary: This review sheds a bright light towards several mechanisms at both cellular and molecular level, correlation of NP-mediated disease mechanism and possible cell surface biomarkers (receptors), and identified genes could be more promising for their pharmacological targets.

Phytocompounds of Onion Target Heat Shock Proteins (HSP70s) to Control Breast Cancer Malignancy.

Globally, breast cancer is one of the leading invasive cancers in women. Moreover, the use of chemotherapeutic drugs for treating cancer produces toxic side effects and has even led to drug resistance. This research paper focuses on targeting three heat shock proteins belonging to 70 kDa subfamily (HSP70s), predominantly, Mortalin, Binding Immunoglobulin Protein (BiP), and Stress Inducible HSP70 (Stress Inducible Heat Shock Protein 70) involved in breast cancer malignancy using different phytocompounds of onion. Phytocompounds of onion (ligands) obtained from different literature sources and the conventional drug, Tamoxifen (standard ligand), used for treating breast cancer are docked against three HSP70s (target proteins) through molecular docking. Molecular docking helps to determine protein-ligand interactions with minimum binding affinity. A comparative analysis revealed that fourteen phytocompounds of onion have lesser binding affinity and formed more stable complexes with the target proteins compared to that of the conventional drug. This evidence can be used and confirmed further through in vitro (cell culture) and in vivo (animal models) studies, and then, these phytocompounds can be modulated efficiently as potential therapeutics for treating breast cancer with less or nearly no side effects. In Silico work represented here targets three heat shock proteins belonging to 70 kDa subfamily (HSP70s)-Mortalin, Binding Immunoglobulin Protein (BiP), and Stress Inducible HSP70 involved in breast cancer malignancy using different phytocompounds of onion to identify potential phytocompounds that can treat breast cancer with nearly no side effects.

Lipidomics: An excellent tool for chronic disease detection.

It has been known as almost all the cells consists a lipid molecule which has a considerable impact in various biological processes. Lipids have been investigated with a potential role for the formation of cellular membrane and thereby maintaining the structural integrity. Omics has placed as a combined technologies utilized for an exploaration of mechanistic actions in several kinds of molecules that make up the cells of an organism. Lipidomics has been recognized as a newly emerged branch of omics technology. This technology has the captivating factors to classify and characterize almost all the cellular lipids with the help of various analytical techniques and computational biological plateform. In lipidomics studies, structural display of several lipid biomarkers could also be analyzed and considered for actual disease diagnosis procedures. This could also replace certain traditional diagnostics method at all over the globe. Our review focuses how important this lipidomics particularly in disease diagnosis and also covers various analytical techniques and computational methods or bioinformatics tools in for the diagnosis of disease. In addtion, we also pinponted the possible role of lipids in several kinds of cellular disorders including cancer, neurodegenerative diseases, cardiovascular diseases, diabetes and obesity in human population. .

SARS-CoV-2 Inhibitors from Nigella Sativa.

The recently encountered severe acute respiratory syndrome coronavirus 2 creates huge predicaments among various countries. Lack of specific treatment of COVID-19 disease demands urgency in drug design against SARS-CoV-2 targets. Nigella sativa the miraculous herb native to South and Southwest Asia and belonging to the family Ranunculaceae, due to its beneficial bioactive properties, was used by us for performing in silico study to analyze the potential of its compounds so that they can target and inhibit SARS-COV-2 proteins including its main protease, the papain-like protease, its helicase, and also the RNA-dependent RNApolymerase, RNA-binding protein, Endoribonuclease, receptor-binding domain, and the RNA-binding domain of nucleocapsid phosphoprotein. The procedure of molecular docking was done with the help of AutoDock-Vina 1.1.2. and along with it the ADMET properties of the best suited ligands were found and Lipinski screening was performed. Among 58 ligands screened, various compounds showed binding energy less than the standard drug chloroquine. Three compounds alpha-hederin, rutin, and nigellamine A2 had the least binding energy with the specific SARS-Cov-2 proteins suggesting their best potential as SARS-CoV-2 inhibitor. Hence, in the future, studies including the in vitro and also the in vivo studies can be carried out for analyzing their true potential and encourage use of nutraceuticals like Nigella sativa to inhibit this virus.

Possible Interaction of Nonsteroidal Anti-inflammatory Drugs Against NF-κB- and COX-2-Mediated Inflammation: In Silico Probe.

In recent years, inflammatory mediators have been considered a possible key for nonsteroidal anti-inflammatory drugs (NSAID's). NSAID's have been known as most promising medication against inflammation and its mediated pain. Inflammation could be recognize as a systemic adaptive stimulation triggered by detrimental stimuli as pathogenic attack and endogenous signals mediated injury inside the cells. In addition, there has been an inflammatory key mechanism involved in disease state. NSAIDs have been compromisingly recommended for targeting specific proteins and/or inflammatory-mediated enzymes including cyclooxygenases (COX). This subsequently inhibits the prostaglandins at the site of inflammation. For the past decades, two forms of the COX enzyme have been implicated as COX-1 expressed in cells and tissues and other COX-2 selectively triggered via proinflammatory cytokines at the site of inflammation and/or injury. In addition, NSAID's have also been implicated for the inhibition of NF-κB pathways, and other relevant proteins considered potent candidates for these drugs. NF-κB has been identified a classical proinflammatory signaling pathway. It has been recognized as a primary target for novel anti-inflammatory drugs. In our results, reports are being confirmed via the probable effects of NSAID's on inflammatory-mediated switches. Several studies were considered to enquire the possible interactions of NSAID's and inflammatory hub. Nevertheless, the exact mechanism is still debatable. In our study, NSAID's and their targeted proteins or molecules caused a convincing pattern. For improvised perception, the binding affinity of NSAID's with inflammatory-mediated proteins was quantified using a molecular docking tool. In addition, we have depicted the complex juncture of hydrogen bonding in targeted proteins with NSAID's. Our in silico investigations have revealed NSAID's as the powerful armor against COX-2- and NF-κB-mediated inflammation.

Cellular and molecular mechanism in neurodegeneration: Possible role of neuroprotectants.

In recent years, neurodegeneration has been recognized as a clinical condition that is characterized by neuronal death, dementia, and gradual diminish of cognitive function, poor body coordination and motor disorders. Several studies deciphering cellular and molecular mechanisms show a promising insight for several kinds of damages including neurodegeneration in central nervous system. In addition, there has been an inflammatory key mechanism involved in neurodegenerative disorders. There is a paucity of literature in both cellular- and molecular-mediated targets in damaged neurons at both in vitro and in vivo research models. It has been notified that CNS has a very restricted magnitude of regeneration. Numerous key factors have also been studied and considered as possible culprit of neurodegeneration. Autophagy is a well-known degradation process wherein vesicular machinery as autophagosome transports cytoplasmic contents to the lysosomes. In earlier reports, a bridging connection between autophagy and its associated mechanism has been established. Natural compounds as a neuro-therapeutics have been recognized in neurodegeneration. In our review, we discuss the mechanisms for the onset and progression in neurodegeneration, via inflammation and autophagic machine available in cellular compartments in CNS. This review also discusses about the neuroprotective efficacy of natural compounds against neurodegeneration episodes displays in neuronal platform.

Multi potent aromatic nano colloid: synthesis, characterization and applications.

In this study the aromatic nanocolloids (CANCs) are synthesized using the noble metal silver by using Citronella extract and confirmed through physio chemical analysis. The synthesised CANCs were evaluated for its antimicrobial activity and antibiofilm activity against pathogenic biofilm forming E. coli. In addition, synthesized CANCs were evaluated for the expression of virulent genes encoding AmpC and CTX-M-15. The results confirmed that CANCs showed effective antimicrobial activity through its bacteriostatic, bactericidal and quorum quencher activity and downregulated CTX-M-15 gene. CANCs were validated as alternate to the commercial fungicides to control plant pathogenic fungi such as A. niger MTCC (281), Fusarium graminearum MTCC (2089) and F. udum MTCC (2204). Furthermore, analysis of CANCs on breast cancer (MCF-7) cells under in vitro condition showed that the cytotoxicity of CANCs is dose dependent. Thus, the multifunctional CANCs can be utilized as potential antimicrobial, antifungal and anticancer agent.

High-throughput, Label-Free Quantitative Proteomic Studies of the Anticancer Effects of Electrical Pulses with Turmeric Silver Nanoparticles: an in vitro Model Study.

Triple negative breast cancer (TNBC) represents 15-20% of the over one million new breast cancer cases occurring each year. TNBC is an aggressive cancer phenotype, with low 5-year survival rates, high 3-year recurrence rates, and increased risk of metastasis. A lack of three commonly exploited hormone receptors renders TNBC resistant to endocrine therapies and lends to its critical absence of viable therapeutic targets. This necessitates the development of alternate and effective novel therapeutic strategies for TNBC. Towards this, our current work seeks to develop the technique of Electrical pulse (EP)-mediated Turmeric silver nanoparticles (TurNP) therapy, known as Electrochemotherapy (ECT), to effectively target TNBC cells. This technique involves the efficient delivery of natural bioactive molecules with anti-cancer effects via a biophysical means. In these experiments, the bioactive molecules are turmeric, a dried rhizome of Curcuma longa that has been used for centuries, both as a dietary supplement and as a medicine in Ayurveda (science of life) in the Indian subcontinent and in traditional Chinese medicine. Our results reveal the combined effect of TurNP + EP treatment in reducing MDA-MB-231 cell viability to as low as 9% at 12 h. Showing biological selectivity, this combination treatment has a substantially lower effect on non-tumorigenic mammary epithelial MCF10A cells (67% viability). To gain mechanistic insights into the actions of TurNP-based ECT treatment, we performed high-throughput, label-free quantitative proteomics studies. Proteomics results indicate that TurNP + EP treatment significantly influenced expression of a diverse list of proteins, including receptors, transcription factors, structural proteins, kinases, and metabolic enzymes. This include the downregulation of 25 proteins in PI3K-Akt signaling pathway (such as GRB2, EGFR, EPHA2, GNB1, GNB2, 14-3-3 family, and Integrin family proteins), and 12 proteins (AKR1A1, ALDOA, ALDOC, PGK1, PGM1, PGAM1, ENO1, ENO2, GAPDH, TPI1, LDHA, and LDHB) in the glycolytic pathway with concomitant reduction in metabolite levels (glucose uptake, and intracellular- lactate, glutamine, and glutamate). Compared to TurNP alone, TurNP + EP treatment upregulated 66 endoplasmic reticulum and 193 mitochondrial proteins, enhancing several processes and pathways, including Pyruvate Metabolism, Tricarboxylic acid (TCA) cycle, and Oxidative Phosphorylation (OXPHOS), which redirected the TNBC metabolism to mitochondria. This switch in the metabolism caused excessive production of H2O2 reactive oxygen species (ROS) to inflict cell death in MDA-MB-231 cells, demonstrating the potency of this treatment.

Biogenic AgNPs synthesized via endophytic bacteria and its biological applications.

In this present study, the endophytic bacteria were isolated from the drought-tolerant ornamental plant Pennisetum setaceum. The biomass of endophytic bacteria was utilized for the biogenic synthesis of silver nanoparticles (AgNPs). The synthesis of AgNPs was confirmed by UV-Visible and FTIR spectroscopy followed by SEM analysis. The antibacterial studies were performed through MIC, MBC, and biofilm assays. Efficacy of AgNPs against the human breast cancer (MCF-7) cells was also tested, and the IC50 was determined by MTT assay. In our study, we have observed that the synthesized AgNPs exhibited a dose-dependent cytotoxicity (1-100 µg/mL) against MCF-7 cells and morphological alterations of the cells were also visualized and the IC50 was observed at 50 µg/mL. The treatment of synthesized AgNPs altered the expression of apoptotic proteins including Bax, Bcl-2, and inflammatory marker COX-2 in MCF-7 cells. To the best of our knowledge, this is the first report that demonstrates the AgNPs from endophytic bacteria isolated from the plant Pennisetum setaceum can induce apoptosis in human breast cancer MCF-7 cells. Our results suggest that AgNPs used in this study can be utilized to control human pathogens and can also be utilized to induce apoptosis in breast cancer cells.

Fungal-mediated synthesis of pharmaceutically active silver nanoparticles and anticancer property against A549 cells through apoptosis.

Generally, fungi have the ability to secrete large amounts of secondary metabolites which have the ability to reduce metal ions to metallic nanoparticles. In this report, silver nanoparticles (AgNPs) were synthesized by using an endophytic fungus isolated from the medicinal plant, Catharanthus roseus (Linn.). The endophytic fungus was identified as Botryosphaeria rhodina based on the ITS sequencing. The synthesized AgNPs were characterized by adopting various high-throughput techniques, scanning electron microscopy (SEM) equipped with energy dispersive X-ray analysis (EDAX), high-resolution transmission electron microscopy (HR-TEM) and UV-Visible spectrophotometer. In vitro anticancer efficacy of AgNPs was tested on A-549 cells. The synthesized AgNPs were effective in scavenging free radicals and induced hallmarks of apoptosis including nuclear and DNA fragmentation in lung (A549) cancer cell lines under in vitro conditions. The results suggested that the natural biomolecules in the endophytic fungi incorporated into the nanoparticles could be responsible for the synergetic cytotoxic activity against cancer cells. The AgNPs were found to have cytotoxicity IC50 of 40 µg/mL against A549 cells. To the best our knowledge, this is the first report demonstrating that AgNPs from Botryosphaeria rhodina could be able to induce apoptosis in various types of cancer cells as a novel strategy for cancer treatment.