Assessment of the mechanistic role of an Indian traditionally used ayurvedic herb Bacopa monnieri (L.)Wettst. for ameliorating oxidative stress in neuronal cells.

ETHNOPHARMACOLOGICAL RELEVANCE: This study has important ethnopharmacological implications since it systematically investigated the therapeutic potential of Bacopa monnieri(L.) Wettst. (Brahmi) in treating neurological disorders characterized by oxidative stress-a growing issue in the aging population. Bacopa monnieri, which is strongly rooted in Ayurveda, has long been recognized for its neuroprotective and cognitive advantages. The study goes beyond conventional wisdom by delving into the molecular complexities of Bacopa monnieri, particularly its active ingredient, Bacoside-A, in countering oxidative stress. The study adds to the ethnopharmacological foundation for using this herbal remedy in the context of neurodegenerative disorders by unravelling the scientific underpinnings of Bacopa monnieri's effectiveness, particularly at the molecular level, against brain damage and related conditions influenced by oxidative stress. This dual approach, which bridges traditional wisdom and modern investigation, highlights Bacopa monnieri's potential as a helpful natural remedy for oxidative stress-related neurological diseases. AIM OF THE STUDY: The aim of this study is to investigate the detailed molecular mechanism of action (in vitro, in silico and in vivo) of Bacopa monnieri (L.) Wettst. methanolic extract and its active compound, Bacoside-A, against oxidative stress in neurodegenerative disorders. MATERIALS AND METHODS: ROS generation activity, mitochondrial membrane potential, calcium deposition and apoptosis were studied through DCFDA, Rhodamine-123, FURA-2 AM and AO/EtBr staining respectively. In silico study to check the effect of Bacoside-A on the Nrf-2 and Keap1 axis was performed through molecular docking study and validated experimentally through immunofluorescence co-localization study. In vivo antioxidant activity of Bacopa monnieri extract was assessed by screening the oxidative stress markers and stress-inducing hormone levels as well as through histopathological analysis of tissues. RESULTS: The key outcome of this study is that the methanolic extract of Bacopa monnieri (BME) and its active component, Bacoside-A, protect against oxidative stress in neurodegenerative diseases. At 100 and 20 µg/ml, BME and Bacoside-A respectively quenched ROS, preserved mitochondrial membrane potential, decreased calcium deposition, and inhibited HT-22 mouse hippocampus cell death. BME and Bacoside-A regulated the Keap1 and Nrf-2 axis and their downstream antioxidant enzyme-specific genes to modify cellular antioxidant machinery. In vivo experiments utilizing rats subjected to restrained stress indicated that pre-treatment with BME (50 mg/kg) downregulated oxidative stress markers and stress-inducing hormones, and histological staining demonstrated that BME protected the neuronal cells of the Cornu Ammonis (CA1) area in the hippocampus. CONCLUSIONS: Overall, the study suggests that Bacopa monnieri(L.) Wettst. has significant potential as a natural remedy for neurodegenerative disorders, and its active compounds could be developed as new drugs for the prevention and treatment of oxidative stress-related diseases.

Redox signalling regulates breast cancer metastasis via phenotypic and metabolic reprogramming due to p63 activation by HIF1α.

BACKGROUND: Redox signaling caused by knockdown (KD) of Glutathione Peroxidase 2 (GPx2) in the PyMT mammary tumour model promotes metastasis via phenotypic and metabolic reprogramming. However, the tumour cell subpopulations and transcriptional regulators governing these processes remained unknown. METHODS: We used single-cell transcriptomics to decipher the tumour cell subpopulations stimulated by GPx2 KD in the PyMT mammary tumour and paired pulmonary metastases. We analyzed the EMT spectrum across the various tumour cell clusters using pseudotime trajectory analysis and elucidated the transcriptional and metabolic regulation of the hybrid EMT state. RESULTS: Integration of single-cell transcriptomics between the PyMT/GPx2 KD primary tumour and paired lung metastases unraveled a basal/mesenchymal-like cluster and several luminal-like clusters spanning an EMT spectrum. Interestingly, the luminal clusters at the primary tumour gained mesenchymal gene expression, resulting in epithelial/mesenchymal subpopulations fueled by oxidative phosphorylation (OXPHOS) and glycolysis. By contrast, at distant metastasis, the basal/mesenchymal-like cluster gained luminal and mesenchymal gene expression, resulting in a hybrid subpopulation using OXPHOS, supporting adaptive plasticity. Furthermore, p63 was dramatically upregulated in all hybrid clusters, implying a role in regulating partial EMT and MET at primary and distant sites, respectively. Importantly, these effects were reversed by HIF1α loss or GPx2 gain of function, resulting in metastasis suppression. CONCLUSIONS: Collectively, these results underscored a dramatic effect of redox signaling on p63 activation by HIF1α, underlying phenotypic and metabolic plasticity leading to mammary tumour metastasis.

Identification and mechanistic exploration of structural and conformational dynamics of NF-kB inhibitors: rationale insights from in silico and in vitro studies.

Increased expression of target genes that code for proinflammatory chemical mediators results from a series of intracellular cascades triggered by activation of dysregulated NF-κB signaling pathway. Dysfunctional NF-kB signaling amplifies and perpetuates autoimmune responses in inflammatory diseases, including psoriasis. This study aimed to identify therapeutically relevant NF-kB inhibitors and elucidate the mechanistic aspects behind NF-kB inhibition. After virtual screening and molecular docking, five hit NF-kB inhibitors opted, and their therapeutic efficacy was examined using cell-based assays in TNF-α stimulated human keratinocyte cells. To investigate the conformational changes of target protein and inhibitor-protein interaction mechanisms, molecular dynamics (MD) simulations, binding free energy calculations together with principal component (PC) analysis, dynamics cross-correlation matrix analysis (DCCM), free energy landscape (FEL) analysis and quantum mechanical calculations were carried out. Among identified NF-kB inhibitors, myricetin and hesperidin significantly scavenged intracellular ROS and inhibited NF-kB activation. Analysis of the MD simulation trajectories of ligand-protein complexes revealed that myricetin and hesperidin formed energetically stabilized complexes with the target protein and were able to lock NF-kB in a closed conformation. Myricetin and hesperidin binding to the target protein significantly impacted conformational changes and internal dynamics of amino acid residues in protein domains. Tyr57, Glu60, Lys144 and Asp239 residues majorly contributed to locking the NF-kB in a closed conformation. The combinatorial approach employing in silico tools integrated with cell-based approaches substantiated the binding mechanism and NF-kB active site inhibition by the lead molecule myricetin, which can be explored as a viable antipsoriatic drug candidate associated with dysregulated NF-kB.Communicated by Ramaswamy H. Sarma.

Pterostilbene-Isothiocyanate Inhibits Proliferation of Human MG-63 Osteosarcoma Cells via Abrogating ß-Catenin/TCF-4 Interaction-A Mechanistic Insight.

Osteosarcoma, a highly metastasizing bone neoplasm, is a leading cause of death and disability in children and adolescents worldwide. Osteosarcoma is only suboptimally responsive to surgery and radio- and chemotherapy, that too with adverse side effects. Hence, there is a necessary need for safer alternative therapeutic approaches. This study evaluated the anticancer effects of the semi-synthetic compound, pterostilbene-isothiocyanate (PTER-ITC), on human osteosarcoma MG-63 cells through cytotoxicity, wound-healing, and transwell-migration assays. Results showed that PTER-ITC specifically inhibited the survival, proliferation, and migration of osteosarcoma cells. PTER-ITC induced apoptosis in MG-63 cells by disrupting mitochondrial membrane potential, as evident from the outcomes of different cytological staining. The antimetastatic potential of PTER-ITC was evaluated through immunostaining, RT-qPCR, and immunoblotting. In silico (molecular docking and dynamic simulation) and, subsequently, biochemical [co-immunoprecipitation (Co-IP) and luciferase reporter] assays deciphered the underlying mode-of-action of this compound. PTER-ITC increased E-cadherin and reduced N-cadherin levels, thereby facilitating the reversal of epithelial-mesenchymal transition (EMT). It also modulated the expressions of proliferative cell nuclear antigen (PCNA), caspase-3, poly [ADP-ribose] polymerase (PARP-1) and matrix metalloproteinase-2/9 (MMPs-2/9) at transcriptional and translational levels. PTER-ITC interfered with the ß-catenin/transcription factor-4 (TCF-4) interaction in silico by occupying the ß-catenin binding site on TCF-4, confirmed by their reduced physical interactions (Co-IP assay). This inhibited transcriptional activation of TCF-4 by ß-catenin (as shown by luciferase reporter assay). In conclusion, PTER-ITC exhibited potent anticancer effects in vitro against human osteosarcoma cells by abrogating the ß-catenin/TCF-4 interaction. Altogether, this study suggests that PTER-ITC may be regarded as a new approach for osteosarcoma treatment.

Pterostilbene-isothiocyanate reduces miR-21 level by impeding Dicer-mediated processing of pre-miR-21 in 5-fluorouracil and tamoxifen-resistant human breast cancer cell lines.

Converging evidences identifies that microRNA-21 (miR-21) is responsible for drug resistance in breast cancer. This study aims to evaluate the miR-21-modulatory potential of a hybrid compound, pterostilbene-isothiocyanate (PTER-ITC), in tamoxifen-resistant MCF-7 (TR/MCF-7) and 5-fluorouracil-resistant MDA-MB 231 (5-FUR/MDA-MB 231) breast cancer cell lines, established by repeated exposure to gradually increasing the concentrations of tamoxifen and 5-fluorouracil, respectively. The outcome of this study shows that PTER-ITC effectively reduced the TR/MCF-7 (IC50: 37.21 µM) and 5-FUR/MDA-MB 231 (IC50: 47.00 µM) cell survival by inducing apoptosis, inhibiting cell migration, colony and spheroid formations in TR/MCF-7 cells, and invasiveness of 5-FUR/MDA-MB 231 cells. Most importantly, PTER-ITC significantly reduced the miR-21 expressions in these resistant cell lines. Moreover, the downstream tumor suppressor target gene of miR-21 such as PTEN, PDCD4, TIMP3, TPM1, and Fas L were upregulated after PTER-ITC treatment, as observed from transcriptional (RT-qPCR) and translational (immunoblotting) data. In silico and miR-immunoprecipitation (miR-IP) results showed reduced Dicer binding to pre-miR-21, after PTER-ITC treatment, indicating inhibition of miR-21 biogenesis. Collectively, the significance of this study is indicated by preliminary evidence for miR-21-modulatory effects of PTER-ITC that highlights the potential of this hybrid compound as an miR-21-targeting therapeutic agent.

A review article on the dosimetry in intraluminal HDR brachytherapy of lung carcinoma.

Lung cancer is one of the most common cancers in the world. Intraluminal brachytherapy (BT) is one of the most adopted treatment modalities for lung malignancies with Ir-192 source in radiotherapy. In intraluminal BT, treatment delivery is required to be very accurate and precise with respect to the plan created in the treatment planning system (TPS). The BT dosimetry is necessary for better treatment outcomes. Therefore in this review article, some relevant studies were identified and analyzed for dosimetric outcomes in intraluminal BT in lung malignancies. The dosimetry in BT for plan verification is not presently in practice, which needs to be performed to check the variation between the planned and measured doses. The necessary dosimetric work done by the various researchers in intraluminal BT such as the Monte Carlo CYLTRAN code was used to calculate and measure the dose rate in any medium. Anthropomorphic phantom was used to measure doses at some distance from the source with Thermo luminescence dosimeters (TLDs). The dosimetric influence of air passage in the bronchus was evaluated with the GEANT4 Monte Carlo method. A pinpoint chamber was used to measure and quantify the impact of inhomogeneity in wax phantom for the Ir-192 source. The Gafchromic films and Monte Carlo methods were used to find the phantom and heterogeneities, which were found to underestimate the dose for the lungs and overestimated for the bones in TPS. The exact tool to quantify the variation in planned and delivered doses should be cost-effective and easy to use possibly with tissue equivalent phantoms and Gafchromic films in lung malignancies treatment.

Angle of concavity in planning target volume can be adopted as selection criteria for intensity-modulated radiation therapy or three-dimensional conformal radiotherapy technique in brain tumors.

Introduction: With innovation of medical imaging, radiotherapy attempts to conform the high dose region to the planning target volume (PTV). The present work aimed to assess the angle of concavity in PTV can be adopted as selection criteria for intensity-modulated radiation therapy (IMRT) or three-dimensional conformal radiotherapy (3DCRT) technique in Brain tumors. Materials and Methods: Thirty previously irradiated patients with brain tumors were replanned with both 3DCRT and IMRT technique. Angle of concavity (dip) in the PTV near the organs at risk was measured in the contoured structure set images of each patient. These cases were divided into three groups where angles were 0°, >120° and <120°. Dose of 60 Gy/30# was fixed. Results: In Group 1, the IMRT plan had better TV95% as compared to 3DCRT respectively with significant P value (P = 0.002). Mean of conformity index (CI) and Homogeneity Index (HI) were comparable. For Group 2 (angle >120°), the IMRT plan had better TV95% as compared to 3DCRT respectively with a significant P value (P = 0.021). HI and CI were not significant. For Group 3 (<120°), IMRT plan had better TV95% as compared to 3DCRT respectively with a significant P value (P = 0.001). HI and CI were better in IMRT arm with significant P value. Conclusion: The results from this study showed that the angle of concavity can be considered as an additional objective tool for selection criteria whether tumor can be treated with IMRT or 3DCRT. Tumors where angle of concavity was <120°, HI and CI provided more uniformity and conformity of dose distribution inside PTV with significant P values.

Gallate Moiety of Catechin Is Essential for Inhibiting CCL2 Chemokine-Mediated Monocyte Recruitment.

Leukocyte recruitment witnesses an orchestrated complex formation between the chemokines and their molecular partners. CCL2 chemokine that regulates monocyte trafficking is a worthwhile system from the pharmaceutical perspective. In the current study, four major catechins (EC/EGC/ECG/EGCG) were assessed for their inhibitory potential against CCL2-regulated monocyte/macrophage recruitment. Interestingly, catechins with the gallate moiety (ECG/EGCG) could only attenuate the CCL2-induced macrophage migration. These molecules specifically bound to CCL2 on a pocket comprising the N-terminal, ß0-sheets, and ß3-sheets, and the binding affinity of ECGC (Kd = 22 ± 4 µM) is ∼4 times higher than that of the ECG complex (Kd = 85 ± 6 µM). MD simulation analysis evidenced that the molecular specificity/stability of CCL2-catechin complexes is regulated by multiple factors, including stereospecificity, number of hydroxyl groups on the annular ring-B, the positioning of the carbonyl group, and the methylation of the galloyl ring. Further, a significant overlap on the binding surface of CCL2 for EGCG/ECG and receptor interactions as evidenced from NMR data provided the rationale for the observed inhibition of macrophage migration in response to EGCG/ECG binding. In summary, these galloylated epicatechins can be considered as potent protein-protein interaction (PPI) inhibitors that regulate CCL2-directed leukocyte recruitment for resolving inflammatory/immunomodulatory disorders.

Promising antivirals for PLpro of SARS-CoV-2 using virtual screening, molecular docking, dynamics, and MMPBSA.

The recent pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (COVID-19) is a viral respiratory disease that has been spread all over the globe. Therefore, it is an urgent requirement to identify and develop drugs for this contagious infection. The papain-like protease (PLpro) of SARS-CoV-2 performs critical functions in virus replication and immune evasion, making it an enticing therapeutic target. SARS-CoV-2 and SARS-CoV PLpro proteases have significant similarities, and an inhibitor discovered for SARS-CoV PLpro is an exciting first step toward therapeutic development. Here, a set of antiviral molecules were screened at the catalytic and S-binding allosteric sites of papain-like protease (PLpro). Molecular docking results suggested that five molecules (44560613, 136277567, S5652, SC75741, and S3833) had good binding affinities at both sites of PLpro. Molecular dynamics analysis like root mean square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration (Rg), solvent accessible surface area (SASA), and hydrogen bond results showed that identified molecules with PLpro tend to form stable PLpro-inhibitor(s) complexes. Molecular Mechanics/Position-Boltzmann Surface Area (MMPBSA) analysis confirmed that antiviral molecules bound PLpro complex had lower energy (-184.72 ± 7.81 to -215.67 ± 6.73 kJ/mol) complexes. Noticeably, computational approaches revealed promising antivirals candidates for PLpro, which may be further tested by biochemical and cell-based assays to assess their potential for SARS-CoV-2.Communicated by Ramaswamy H. Sarma.

Definitive chemoradiation in nonmetastatic squamous cell carcinoma anal canal: A single-institution experience.

Purpose: The purpose of the study was to analyze the survival outcomes and toxicities in squamous cell carcinoma anal canal treated with definitive chemoradiotherapy. Materials and Methods: Retrospective analysis of 51 patients with squamous cell carcinoma anal canal treated with chemoradiotherapy was done. Data were collected and analyzed for disease-free survival (DFS), colostomy-free survival (CFS), overall survival (OS), and acute/late toxicities. Results: Out of total 51 patients, only 44 patients had a follow-up of more than 36 months and were analyzed. After a median follow-up of 46 months (range 10-68 months), the 3-year DFS was 73.9%. Three patients developed locoregional recurrence, while one patient developed distant metastasis. At 3-year OS rate was 77%. Out of 44 patients, six patients lost to follow-up, while two patients died due to progressive disease and two due to noncancer causes. 3-year CFS rate was 59%. Most common grade >3 acute toxicities were skin reactions in nine (18%), followed by hematological in eight (16%) patients. Conclusion: Definitive chemoradiotherapy in anal canal results in good oncological outcomes with sphincter preservation. No severe treatment-related toxicities were observed.

Pterostilbene-isothiocyanate impedes RANK/TRAF6 interaction to inhibit osteoclastogenesis, promoting osteogenesis in vitro and alleviating glucocorticoid induced osteoporosis in rats.

Prolonged glucocorticoid treatment often leads to glucocorticoid-induced osteoporosis (GIOP), a common iatrogenic complication. This study has explored the anti-osteoporotic potential of semi-synthetic compound, pterostilbene isothiocyanate (PTER-ITC) in GIOP rat model and bone formation potential in vitro. Dysregulated bone-remodelling leads to osteoporosis. PTER-ITC has shown anti-osteoclastogenic activity in vitro. However, its molecular target remains unidentified, which has been explored in this study through in silico and experimental approaches. Alizarin Red S and von-Kossa staining, and alkaline phosphatase (ALP) activity showed the osteogenic differentiation potential of PTER-ITC in pre-osteoblastic mouse MC3T3-E1 and human hFOB 1.19 cells, further, confirmed through the expressions of osteogenic markers at transcriptional (RT-qPCR) and translational (immunoblotting) levels. The anti-osteoclastogenic property of PTER-ITC was confirmed through inhibition of actin ring formation in mouse RAW 264.7 and human THP-1 macrophagic cells. Molecular docking and molecular dynamic simulation showed that PTER-ITC inhibited the crucial osteoclastogenic RANK/TRAF6 interaction, which was further confirmed biochemically through co-immunoprecipitation assay. Osteoporotic bone architecture [validated through scanning electron microscopy (SEM), X-ray radiography, and micro-computed tomography (µ-CT)], physiology (confirmed through compression testing, Young's modulus and stress versus strain output) and histology (verified through hematoxylin-eosin, Alizarin Red S, von-Kossa and Masson-trichrome staining) of PTER-ITC-treated GIOP female Wistar rats were assuaged. Osteoporotic amelioration through PTER-ITC treatment was further substantiated through serum biomarkers, like, parathyroid hormone (PTH), ALP, calcium (Ca2+), Procollagen type I N-terminal propeptide (P1NP), and 25-hydroxy vitamin D. In conclusion, this study identifies the molecular target of PTER-ITC in impeding osteoclastogenesis and facilitating osteogenesis to ameliorate osteoporosis.

Screening of Severe Acute Respiratory Syndrome Coronavirus 2 RNA-Dependent RNA Polymerase Inhibitors Using Computational Approach.

The detrimental effect of coronavirus disease 2019 (COVID-19) pandemic has manifested itself as a global crisis. Currently, no specific treatment options are available for COVID-19, so therapeutic interventions to tackle the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection must be urgently established. Therefore, cohesive and multidimensional efforts are required to identify new therapies or investigate the efficacy of small molecules and existing drugs against SARS-CoV-2. Since the RNA-dependent RNA Polymerase (RdRP) of SARS-CoV-2 is a promising therapeutic target, this study addresses the identification of antiviral molecules that can specifically target SARS-CoV-2 RdRP. The computational approach of drug development was used to screen the antiviral molecules from two antiviral libraries (Life Chemicals [LC] and ASINEX) against RdRP. Here, we report six antiviral molecules (F3407-4105, F6523-2250, F6559-0746 from LC and BDG 33693278, BDG 33693315, LAS 34156196 from ASINEX), which show substantial interactions with key amino acid residues of the active site of SARS-CoV-2 RdRP and exhibit higher binding affinity (>7.5 kcalmol-1) than Galidesivir, an Food and Drug Administration-approved inhibitor of the same. Further, molecular dynamics simulation and Molecular Mechanics Poisson-Boltzmann Surface Area results confirmed that identified molecules with RdRP formed higher stable RdRP-inhibitor(s) complex than RdRP-Galidesvir complex. Our findings suggest that these molecules could be potential inhibitors of SARS-CoV-2 RdRP. However, further in vitro and preclinical experiments would be required to validate these potential inhibitors of SARS-CoV-2 protein.

Study of dosimetry and clinical factors for assessment of xerostomia in head and neck squamous cell carcinoma treated by intensity-modulated radiotherapy: A prospective study.

AIM: Clinical and dosimetric factors related to toxicity in terms of xerostomia in patients with head and neck squamous cell cancer (HNSCC) treated with intensity-modulated radiotherapy (IMRT). MATERIALS AND METHODS: Patients older than 18 years, with the WHO Performance Status Score <2 with primary diagnosis of HNSCC Stage II, III, and IV who had undergone primary or postoperative radiotherapy (RT) treated by IMRT at the center, from November 2015 to November 2016 were included in the study. Patients were assessed by physical examination and questioned to score their quality of life for dryness (HNDR) and stickiness (HNSS) by EORTC-HN-35 (Hindi or English version) at baseline (before treatment), at 3, 6, and 12 months following treatment. The validation of EORTC-HN-35 for HNDR and HNSS in patients was handed. RESULTS: Thirty patients were included in the study. The mean symptom score values for HNSS at baseline, 3, 6, and 12 months' post-RT treatment were 17.8, 62.2, 64.4, and 20.8, respectively. Dryness and stickiness also increased over 3-6 months in follow-up but slightly relieved at 12 months, but it could not reach to baseline. In subgroup analysis, at baseline mean score of dryness of mouth in elderly patients (≥60 years) (P = 0.248), poor performance status (Eastern Cooperative Oncology Group 2) (P = 0.80) and patients with advanced stage (Stage III and IVA) (P = 0.185) was higher. Correlation of normal tissue complication probability for xerostomia with contralateral mean parotid gland showed insignificant linearity with shallow curve. CONCLUSION: Patients remained symptomatic for xerostomia chiefly till 6 months' postirradiation, but it was slightly relieved in 12 months but could not reach the baseline. Dosimetric sparing ofcontralateral parotid resulted in decreased probability of developing xerostomia.

Pterostilbene-isothiocyanate inhibits breast cancer metastasis by selectively blocking IKK-ß/NEMO interaction in cancer cells.

Metastasis, the main cause of breast cancer-associated fatalities, relies on many regular pathways involved in normal cell physiology and metabolism, thus, making it challenging to identify disease-specific therapeutic target(s). Chemically synthesized anti-metastatic agents are preferred for their fast and robust actions. However, these agents have adverse side effects, thus, increasingly favouring the identification of phytocompounds as suitable alternatives. Resveratrol and pterostilbene have long been established as potent anti-cancer agents. Earlier studies from our laboratory documented the anti-cancer activities associated with pterostilbene-isothiocyanate (PTER-ITC), a derivative of pterostilbene. The current study focuses on evaluating the anti-metastatic property of PTER-ITC and the underlying mechanism, by employing in silico, in vitro, and in vivo approaches. The significant anti-metastatic activity of PTER-ITC was observed in vitro against breast cancer metastatic cell line (MDA-MB-231) and in vivo in the 4T1 cell-induced metastatic mice model. Epithelial-mesenchymal transition (EMT), a hallmark of metastasis regulated by the transcription factors, Snail1 and Twist, was found to be reverted in vitro by PTER-ITC treatment. PTER-ITC blocked the activation of NF-κB/p65 and its concomitant nuclear translocation, resulting in the transcriptional repression of its target genes, Snail1 and Twist. PTER-ITC prevented the formation of IKK complex, central to NF-κB activation, by binding to the NEMO-binding domain (NBD) of IKK-ß and inhibiting its interaction with NEMO (NF-κB essential modulator). According to our observations, PTER-ITC attenuated NF-κB activation selectively in cancerous cells. In conclusion, this study demonstrated that PTER-ITC is a potent anti-metastatic agent capable of targeting physiologically important pathways in a cancer-specific manner.

Nutraceutical regulation of miRNAs involved in neurodegenerative diseases and brain cancers.

The human brain is a well-connected, intricate network of neurons and supporting glial cells. Neurodegenerative diseases arise as a consequence of extensive loss of neuronal cells leading to disruption of their natural structure and function. On the contrary, rapid proliferation and growth of glial as well as neuronal cells account for the occurrence of malignancy in brain. In both cases, the molecular microenvironment holds pivotal importance in the progression of the disease. MicroRNAs (miRNA) are one of the major components of the molecular microenvironment. miRNAs are small, noncoding RNAs that control gene expression post-transcriptionally. As compared to other tissues, the brain expresses a substantially high number of miRNAs. In the early stage of neurodegeneration, miRNA expression upregulates, while in oncogenesis, miRNA expression is gradually lost. Neurodegeneration and brain cancer is presumed to be under the influence of identical pathways of cell proliferation, differentiation and cell death which are tightly regulated by miRNAs. It has been confirmed experimentally that miRNA expression can be regulated by nutraceuticals - macronutrients, micronutrients or natural products derived from food; thereby making dietary supplements immensely significant for targeting miRNAs having altered expression patterns during neurodegeneration or oncogenesis. In this review, we will discuss in detail, about the common miRNAs involved in brain cancers and neurodegenerative diseases along with the comprehensive list of miRNAs involved separately in both pathological conditions. We will also discuss the role of nutraceuticals in the regulation of those miRNAs which are involved in both of these pathological conditions.

Study of volumetric and dosimetric changes during fractionated radiotherapy in head and neck cancers: A single centre experience.

AIMS AND OBJECTIVE: The assessment of volumetric and dosimetric changes in the head-and-neck cancer during fractionated radiotherapy by intensity-modulated radiotherapy (IMRT) technique. MATERIALS AND METHODS: A single-center prospective observational hospital-based study with a sample size of 20 cases of the head-and--neck squamous cell carcinoma over 1 year treated with chemoradiotherapy 66-70 Gy/33-35#@2 Gy/fraction with weekly cisplatin 35 mg/m2. After contouring of target volumes (TVs) and organs at risk (OARs) in initial computed tomography (CT) scan, all patients were planned and treated by the IMRT technique. We re-delineated the TVs and OARs in the second (CT15#) and third (CT30#) planning CT scan, and the initial plan was implemented in the re-CT scan dataset with the same optimization and doses. The volumetric and dosimetric changes during fractionated radiotherapy of TVs and OARs were evaluated and compared. Nonparametric Wilcoxon-signed-rank test was used to compare the means between each plan. RESULTS: For all 20 patients, plans were compared for volumetric and dosimetric parameters on repeat CT scans. The mean variation in gross tumor volume (GTV) and planning TV (PTV) was significant after 15 and 30 fractions of radiotherapy. On dosimetric evaluation, there was a significant increase in doses to GTV and OARs (parotid, spinal cord, and cochlea) with a significant P value. However, doses to the OARs were not exceeded the maximum tolerance limit. CONCLUSION: This prospective single-center study concluded that two repeat imaging, along with re-planning improved TV coverage and decreased doses to the normal tissue. Larger studies with more sample sizes are required to set the criteria for replanning.

Selective Release of Doxorubicin from Cucurbit[8]uril Stabilized Gold Supra-Pyramid Host at pH of Small Intestine.

Gold supra-pyramid structures were obtained by the addition of acidic solution of cucurbit[8]uril (CB[8]) to an aqueous solution of citrate stabilized gold nanoparticles (AuNP). The reaction resulted in the precipitation of supra-pyramid from the solution after just 1 min of shaking. Microscopic images confirmed formation of the supra-pyramid. The stepwise structural transformation towards the supra-pyramid was examined with variable concentrations of CB[8] to AuNP solution. Anionic counter parts of these acids (Br- , NO3 - , SO4 2- and Cl- ) controlled the size of the synthesized supra-pyramids. These supra-pyramid hosts showed uptake of three anticancer drugs: oral drugs etoposide, prednisolone and intravenous drug doxorubicin. Releases of drugs from these hosts were emulated at acidic stomach pH, basic small intestinal pH and in the presence of human serum albumin (HSA). The specific release of doxorubicin was confirmed at small intestinal pH 7.4. Poor release of drugs in presence of CB[8] specific guest 1-adamantanamine confirmed the role of the supra-pyramid as the exclusive host. The release of doxorubicin from the supra-pyramid at pH 7.4 was confirmed by fluorescence microscopic imaging with prostate cancer DU-145 cell line.

Anisotropically Conductive Biodegradable Scaffold with Coaxially Aligned Carbon Nanotubes for Directional Regeneration of Peripheral Nerves.

Fascicular rearrangement of an injured peripheral nerve requires reconnection of nerve sprouts from anterior and Büngner bands from distal sides of the lesion, failing to which leads to inefficient regeneration of the injured nerve. However, existing neural scaffolds have limited neuroregeneration efficiency because of either the lack of alignment of fibers and a conductive second phase, leading to compromised electrical conductivity, or the lack of extracellular matrix components and in vivo validation. The present study reports a biocompatible, multiwall carbon nanotube (MWCNT)-reinforced, anisotropically conductive, electrospun, aligned nanofibrous scaffold, ensuring maximal peripheral nerve regeneration. Electrospinning parameters were modulated to deposit random and parallel fibers in separate scaffolds for comparative analysis on the effect of fiber alignment on regeneration. Both types of scaffolds were reinforced with MWCNTs to impart electrical conductivity. Nonreinforced scaffolds were nonconductive. In this comparative study, MWCNT-reinforced, aligned scaffolds showed better tensile property with increased conductivity along the direction of alignment, thereby ensuring an escalated neural-regeneration rate. Collectively, in vitro studies established the scaffolds to be highly biocompatible, promoting cell growth and proliferation. With 85% more anisotropic conductivity in the direction of the alignment and the degradation kinetics tuned to the regeneration regime, the MWCNT-reinforced, aligned scaffold efficiently healed injured sciatic nerves in rats within 30 days. Rigorous revivification of the tissue was due to coordinated Wallerian degeneration and expedited guided axonal regeneration. Structural and functional analysis of nerves in vivo showed the aligned, MWCNT-reinforced scaffold to be very efficient in peripheral sciatic nerve regeneration. This study notes the efficacy of the coaxially aligned, MWCNT-reinforced neural scaffold, with a capability of establishing remarkable advancement in the field of peripheral neural regeneration.

The Evolving Neural Tissue Engineering Landscape of India.

The healthcare sector in India is witnessing unprecedented advancement. Tissue engineering has become an integral part of healthcare and medicine, particularly where treatments involve functional restoration of any injured or deceased part of the body. Not falling behind much with the progressing medical and healthcare sector of India, tissue engineering is also gaining momentum in the country. Out of several arenas of tissue engineering, India has made its mark in orthopedic and bone regeneration, cosmetic and skin regeneration, and very importantly neural regeneration. There are several articles reviewing the progress and prospects of orthopedic and skin regeneration research in India. However, there is no systematic review on progress, prospects, and pitfalls associated with neural tissue engineering in Indian context. The existing ones mainly focus on the technical advancements in the field from a global perspective. Therefore, it is worthwhile to have an organized look at the evolving neural tissue engineering landscape of India. This review will walk the readers systematically through different aspects of the topic. The review starts with an introduction to the nervous system to help readers appreciate the complexity that must be dealt with while engineering neural tissue. This is followed with a global picture of the neural tissue engineering, prominent research groups working on neural tissue engineering in India, factors that have and are currently molding the prospects of this field, and concluding with an overall perspective on present and future of neural tissue engineering in India.

TiO2 doped chitosan/hydroxyapatite/halloysite nanotube membranes with enhanced mechanical properties and osteoblast-like cell response for application in bone tissue engineering.

The current therapeutic strategies for healing bone defects commonly suffer from the occurrence of bacterial contamination on the graft, resulting in nonunion in the segmental bone defects and the requirement for secondary surgery to remove or sterilize the primary graft. A membrane with enhanced anti-bacterial efficacy, mechanical strength and osteoconductivity would represent an improvement in the therapeutic strategy for guided bone regeneration. The present study aims to optimize the content of halloysite nanotubes (HNTs) and TiO2 in the polymer matrix of chitosan (CTS) with a constant amount of nano-hydroxyapatite (5%) with the objective of mimicking the mechanical and biological microenvironment of the natural bone extracellular matrix with enhanced anti-bacterial efficacy. HNTs are a low-cost alternative to MWNCTs for enhancing the mechanical properties and anti-bacterial efficacy of the composite. From the first stage of the study, it was concluded that the membranes possessed enhanced mechanical properties and optimum biological properties at 7.5% (w/w) loading of HNTs in the composite. In the second stage of this investigation, we studied the effect of the addition of TiO2 nanoparticles (NPs) and TiO2 nanotubes (NTs) in small amounts to the CTS/n-HAP/HNT nanocomposite at 7.5% HNT loading, with an aim to augment the anti-bacterial efficacy and osteoconductivity of this mechanically strong membrane. The study revealed a significant enhancement in the anti-bacterial efficacy, osteoblast-like MG-63 cell proliferation and ALP expression with the addition of TiO2 NTs. The CHH-TiT membrane successfully inhibited the S. aureus and E. coli growth within 16 hours and simultaneously assisted the enhanced proliferation of osteoblast-like cells on its surface. The study supports the potential exploitation of CHH-TiT (7.5% HNT & 0.2% TiO2 NT) membranes as a template for guided bone tissue regeneration.