Mishra, Amit Kumar

Background/Aims: Cells require regular maintenance of proteostasis. Synthesis of new polypeptides and elimination of damaged or old proteins is an uninterrupted mechanism essential for a healthy cellular environment. Impairment in the removal of misfolded proteins can disturb proteostasis; such toxic aggregation of misfolded proteins can act as a primary risk factor for neurodegenerative diseases and imperfect ageing. The critical challenge is to design effective protein quality control (PQC) based molecular tactics that could potentially eliminate aggregation-prone protein load from the cell. Still, targeting specific components of the PQC pathway for the suppression of proteotoxic insults retains several challenges. Earlier, we had observed that LRSAM1 promotes the degradation of aberrant proteins. Here, we examined the effect of resveratrol, a stilbenoid phytoalexin compound, treatment on LRSAM1 E3 ubiquitin ligase, involved in the spongiform neurodegeneration. Methods: In this study, we reported induction of mRNA and protein levels of LRSAM1 in response to resveratrol treatment via RT-PCR, immunoblotting, and immunofluorescence analysis. The LRSAM1-mediated proteasomal-based clearance of misfolded proteins was also investigated via proteasome activity assays, immunoblotting and immunofluorescence analysis. The increased stability of LRSAM1 by resveratrol was demonstrated by cycloheximide chase analysis. Results: Here, we show that resveratrol treatment induces LRSAM1 E3 ubiquitin ligase expression levels. Further, our findings suggest that overexpression of LRSAM1 significantly elevates proteasome activities and improves the degradation of bona fide heat-denatured luciferase protein. Exposure of resveratrol not only slows down the turnover of LRSAM1 but also effectively degrades abnormal proteinaceous inclusions, which eventually promotes cell viability. Conclusion: Our findings suggest that resveratrol facilitates LRSAM1 endogenous establishment, which consequently promotes the proteasome machinery for effective removal of intracellular accumulated misfolded or proteasomal-designated substrates. Altogether, our study proposes a promising molecular approach to specifically trigger PQC signaling for efficacious rejuvenation of defective proteostasis via activation of overburdened proteolytic machinery.

Nanocatalysts are extremely crucial for the expedited synthesis of various chemicals, fuels, and pharmaceutical molecules both in academia and industry. To overcome the limitations of nanocatalysts and or microstructure supported catalysts such as agglomeration (due to inter-particle dipolar forces preventing longer shelf-lives), compromised catalytic activity (e.g., nickel-titanium dioxide bimetallic catalyst, showed high selectivity to hydrogenate 3-nitrostyrene into 3-vinylaniline (90.2 %) compared to unmodified nickel (55.3 %), due to metal-plane formation by titanium dioxide), cytotoxicity (with over 90 % cell killing in the presence of the nanocatalysts above ∼ 0.2 mg/mL), catalyst retrieval (demanding energy intensive procedures such as centrifugation (∼10,000 g and above), membrane filtrations (∼0.2 µm), magnetic separations (0.9–1.1 T) and absurd practical implementation there is a tremendous development of 3-dimensional wide-area supported catalysts. This review update the readers on the evolution of highly catalytic nanoparticles for various heterogeneous catalysis. Uniquely, wide-area supported catalysts wherein the nanoparticles are grafted to 3-dimensional nature-inspired or pristine natural materials as sustainable strategies are discussed. The role of wide-area of the support in overcoming the limitations of nanocatalysts and microstructures by enabling bidirectional reactant access, catalyst efficiency, reusability, stability and sustainability are highlighted. Next, we focus on the metal-affinity and redox-potential of the natural support that aid autogenic biosynthesis and self-assembly of nanocatalysts. Followed by discussions on supplementary properties of the support such as type, structural-hierarchy, surface-area, absorption, porosity and rigidity in tuning the stability, biodegradability, compatibility, functionality and performance of the catalyst. Accentuated, with the impact of support in dictating the choice of fixed batch vs continuous flow reactors, co-relative to modulating the catalytic efficiency and turnover frequencies. Finally, the exclusive role of wide-area of the support and its biological nature in allowing the extraction of noble precursor off the support after catalyst poisoning is emphasized. These discussions, for the first time, spotlight the versatility, resilient nature of the emerging ultra-efficient wide-area supported catalysts that are generated using sustainable procedures for diverse large-volume heterogeneous catalysis.

SCLC is refractory to conventional therapies; targeted therapies and immunological checkpoint inhibitor (ICI) molecules have prolonged survival only marginally. In addition, ICIs help only a subgroup of SCLC patients. Different types of kinases play pivotal roles in therapeutics-driven cellular functions. Therefore, there is a significant need to understand the roles of kinases in regulating therapeutic responses, acknowledge the existing knowledge gaps, and discuss future directions for improved therapeutics for recalcitrant SCLC. Here, we extensively review the effect of dysregulated kinases in SCLC. We further discuss the pharmacological inhibitors of kinases used in targeted therapies for recalcitrant SCLC. We also describe the role of kinases in the ICI-mediated activation of antitumor immune responses. Finally, we summarize the clinical trials evaluating the potential of kinase inhibitors and ICIs. This review overviews dysregulated kinases in SCLC and summarizes their potential as targeted therapeutic agents. We also discuss their clinical efficacy in enhancing anticancer responses mediated by ICIs.

Background: Helicobacter pylori is a prominent causative agent of gastric ulceration, gastric adenocarcinoma and gastric lymphoma and have been categorised as a group 1 carcinogen by WHO. The treatment of H. pylori with proton pump inhibitors and antibiotics is effective but also leads to increased antibiotic resistance, patient dissatisfaction, and chances of reinfection. Therefore, an effective vaccine remains the most suitable prophylactic option for mass administration against this infection. Results: We modelled a multi-chimera subunit vaccine candidate against H. pylori by screening its secretory/outer membrane proteins. We identified B-cell, MHC-II and IFN-γ-inducing epitopes within these proteins. The population coverage, antigenicity, physiochemical properties and secondary structure were evaluated using different in-silico tools, which showed it can be a good and effective vaccine candidate. The 3-D construct was predicted, refined, validated and docked with TLRs. Finally, we performed the molecular docking/simulation and immune simulation studies to validate the stability of interaction and in-silico cloned the epitope sequences into a pET28b(+) plasmid vector. Conclusion: The multiepitope-constructed vaccine contains T- cells, B-cells along with IFN-γ inducing epitopes that have the property to generate good cell-mediated immunity and humoral response. This vaccine can protect most of the world’s population. The docking study and immune simulation revealed a good binding with TLRs and cell-mediated and humoral immune responses, respectively. Overall, we attempted to design a multiepitope vaccine and expect this vaccine will show an encouraging result against H. pylori infection in in-vivo use.

Drug-resistant tuberculosis (TB) outbreak has emerged as a global public health crisis. Therefore, new and innovative therapeutic options like host-directed therapies (HDTs) through novel modulators are urgently required to overcome the challenges associated with TB. In the present study, we have investigated the anti-mycobacterial effect of 4-(Benzyloxy)phenol. Cell-viability assay asserted that 50 μM of 4-(Benzyloxy)phenol was not cytotoxic to phorbol 12-myristate 13-acetate (PMA) differentiated THP-1 (dTHP-1) cells. It was observed that 4-(Benzyloxy)phenol activates p53 expression by hindering its association with KDM1A. Increased ROS, intracellular Ca2+ and phagosome-lysosome fusion, were also observed upon 4-(Benzyloxy)phenol treatment. 4-(Benzyloxy)phenol mediated killing of intracellular mycobacteria was abrogated in the presence of specific inhibitors of ROS, Ca2+ and phagosome-lysosome fusion like NAC, BAPTA-AM, and W7, respectively. We further demonstrate that 4-(Benzyloxy)phenol mediated enhanced ROS production is mediated by acetylation of p53. Blocking of p53 acetylation by Pifithrin-α (PFT- α) enhanced intracellular mycobacterial growth by blocking the mycobactericidal effect of 4-(Benzyloxy)phenol. Altogether, the results showed that 4-(Benzyloxy)phenol executed its anti-mycobacterial effect by modulating p53-mediated ROS production to regulate phagosome-lysosome fusion through Ca2+ production.

Curcumin exhibits a broad spectrum of beneficial health properties that include anti-tumor and anti-cancer activities. The down-regulation of c-myc transcription via stabilizing the G-quadruplex structure formed at the promoter region of the human c-myc gene allows the repression in cancer growth. Small molecules can bind and stabilize this structure to provide an exciting and promising strategy for anti-cancer therapeutics. Herein, we investigated the interaction of Curcumin and its synthetic analogs with G-quadruplex DNA formed at the c-myc promoter by using various biophysical and biochemical assays. Further, its cytotoxic effect and mechanistic insights were explored in various cancer cell lines as well as in multicellular tumor spheroid (MCTS) model. The MCTS possesses almost similar microenvironment as avascular tumors, and micro-metastases can be used as a suitable model for the small molecule-based therapeutics development. Our study provides an expanded overview of the anti-cancer effect of a new Curcumin analog via targeting G-quadruplex structures formed at the promoter region of the human c-myc gene.

The physiochemical properties such as (i) electrical conductivity in solution phase, (ii) response to electrochemical perturbations, (iii) functional group finger print in Fourier Transform InfraRed (FTIR), Ultra Violet-Visible (UV-vis) spectroscopy and (iv) surface morphology employing Scanning Electron Microscopy (SEM) with Energy Dispersive Spectroscopy (EDS) were studied for α,β-tubulin dimer (microtubules) extracted from the plant source, Arachis Hypogaea. To understand the difference in basic mechanism of electronic conduction, a comparison of physiochemical properties of microtubules, Graphite Encapsulated Iron Cobalt nanoparticles (GE-FeCo NPs) and Multiwalled Carbon Nanotubes (MWCNTs) dispersed in the buffer solution were also studied. The electrical conductivity of the microtubules suspended in buffer solution is calculated as 0.056 S/m ± 0.005 S/m and the calculated value is approximately 37 times higher than the value of 0.0015S/m reported in the literature.

Cells regularly synthesize new proteins to replace old and abnormal proteins for normal cellular functions. Two significant protein quality control pathways inside the cellular milieu are ubiquitin proteasome system (UPS) and autophagy. Autophagy is known for bulk clearance of cytoplasmic aggregated proteins, whereas the specificity of protein degradation by UPS comes from E3 ubiquitin ligases. Few E3 ubiquitin ligases, like C-terminus of Hsc70-interacting protein (CHIP) not only take part in protein quality control pathways, but also plays a key regulatory role in other cellular processes like signaling, development, DNA damage repair, immunity and aging. CHIP targets misfolded proteins for their degradation through proteasome, as well as autophagy; simultaneously, with the help of chaperones, it also regulates folding attempts for misfolded proteins. The broad range of CHIP substrates and their associations with multiple pathologies make it a key molecule to work upon and focus for future therapeutic interventions. E3 ubiquitin ligase CHIP interacts and degrades many protein inclusions formed in neurodegenerative diseases. The presence of CHIP at various nodes of cellular protein-protein interaction network presents this molecule as a potential candidate for further research. In this review, we have explored a wide range of functionality of CHIP inside cells by a detailed presentation of its co-chaperone, E3 and E4 enzyme like functions, with central focus on its protein quality control roles in neurodegenerative diseases. We have also raised many unexplored but expected fundamental questions regarding CHIP functions, which generate hopes for its future applications in research, as well as drug discovery.

Background Helminth infections are a global health menace affecting 24% of the world population. They continue to increase global disease burden as their unclear pathology imposes serious challenges to patient management. Neurocysticercosis is classified as neglected tropical disease and is caused by larvae of helminthic cestode Taenia solium. The larvae infect humans and localize in central nervous system and cause NCC; a leading etiological agent of acquired epilepsy in the developing world. The parasite has an intricate antigenic makeup and causes active immune suppression in the residing host. It communicates with the host via its secretome which is complex mixture of proteins also called excretory secretory products (ESPs). Understanding the ESPs interaction with host can identify therapeutic intervention hot spots. In our research, we studied the effect of T. solium ESPs on human macrophages and investigated the post-translation switch involved in its immunopathogenesis. Methodology T. solium cysts were cultured in vitro to get ESPs and used for treating human macrophages. These macrophages were studied for cellular signaling and miR expression and quantification at transcript and protein level. Conclusion We found that T. solium cyst ESPs treatment to human macrophages leads to activation of Th2 immune response. A complex cytokine expression by macrophages was also observed with both Th1 and Th2 cytokines in milieu. But, at the same time ESPs modulated the macrophage function by altering the host miR expression as seen with altered ROS activity, apoptosis and phagocytosis. This leads to activated yet compromised functional macrophages, which provides a niche to support parasite survival. Thus T. solium secretome induces Th2 phenomenon in macrophages which may promote parasite’s survival and delay their recognition by host immune system.

H₂S is recognized as a toxic gas, renowned for its capacity to inflict significant harm upon the respiratory and nervous systems. Consequently, the development of high-performance H₂S sensors holds significant importance. However, traditional fabrication methods such as brush painting and drop casting often yield sensors with inconsistent batch responses due to the unpredictable film formation process, hindering their mass industrial production. Here, we have devised a novel approach to fabricate highly sensitive and selective H₂S sensors utilizing Pd-anchored CuO/SnO₂ heterostructures thin films. An investigation was conducted to analyze the impact of different noble metals (Pt, Au, Ag, and Pd) on CuO/SnO₂ heterostructure thin films concerning their response to H₂S. These films were synthesized through RF sputtering and subsequently decorated with varying durations of CuO (15, 30, 45, 60, and 75 s) and Pd nanoparticles (3, 6, 9, and 12 s) using a sputtering process. Notably, a sputtering time of 60 s for CuO and 9 seconds for Pd significantly enhanced the H₂S sensing performance and selectivity over other gases. The Pd-anchored CuO/SnO₂ thin films revealed an exceptional result of 75.45% to 100 ppm H₂S, demonstrating a detection capability down to 0.5 ppm. These noteworthy outcomes were attained under optimal operating conditions at a temperature of 150 °C. This innovative fabrication technique holds promise for the advancement of gas sensor technology, enabling the creation of portable sensor prototypes suitable for real-time sensing applications.