Environmentally friendly and efficient TBHP-mediated catalytic reaction for the synthesis of substituted benzimidazole-2-ones: In-silico approach to pharmaceutical applications.

A novel method was used to synthesize benzimidazole-2-ones from the corresponding benzimidazolium salts. These salts were subsequently reacted with potassium tertiary butoxide (KOtBu), followed by oxidation using tertiary butyl hydrogen peroxide (TBHP) at room temperature in tetrahydrofuran (THF) to obtain the desired products in 1 h with excellent yields. After optimizing the reaction conditions, the study focused on preparing benzimidazole-2-ones with diverse substituents at N1 and N3 positions, including benzyl, 2',4',6'-trimethyl benzyl groups, and long-chain aliphatic substituents (hexyl, octyl, decyl, and dodecyl). The compounds were characterized by 1H and 13C NMR spectra, of which compound 2a is supported by single crystal XRD. Benzimidazole-2-one compounds exhibited promising anti-inflammatory and anti-cancer properties. The inhibition of mitochondrial Heat Shock Protein 60 (HSP60) of title compounds was also explored. Computational simulations were employed to assess anti-cancer properties of 19 benzimidazole-2-one derivatives (potential drugs). In-silico docking studies demonstrated promising binding interactions with HSP60, and these results were supported by molecular dynamics simulations. Notably, molecules 2b and 2d exhibited high affinity for HSP60 protein, highlighting their potential efficacy. The developed ligands were viable for the treatment of hepatocellular carcinoma (HCC). The findings provide valuable initial evidence supporting the efficacy of benzimidazole-2-ones as HSP60 inhibitors and lay the foundation for subsequent studies, including in-vitro assays.

Master Transcription Factor Reprogramming Unleashes Selective Translation Promoting Castration Resistance and Immune Evasion in Lethal Prostate Cancer.

Signaling rewiring allows tumors to survive therapy. Here we show that the decrease of the master regulator microphthalmia transcription factor (MITF) in lethal prostate cancer unleashes eukaryotic initiation factor 3B (eIF3B)-dependent translation reprogramming of key mRNAs conferring resistance to androgen deprivation therapy (ADT) and promoting immune evasion. Mechanistically, MITF represses through direct promoter binding eIF3B, which in turn regulates the translation of specific mRNAs. Genome-wide eIF3B enhanced cross-linking immunoprecipitation sequencing (eCLIP-seq) showed specialized binding to a UC-rich motif present in subsets of 5' untranslated regions. Indeed, translation of the androgen receptor and major histocompatibility complex I (MHC-I) through this motif is sensitive to eIF3B amount. Notably, pharmacologic targeting of eIF3B-dependent translation in preclinical models sensitizes prostate cancer to ADT and anti-PD-1 therapy. These findings uncover a hidden connection between transcriptional and translational rewiring promoting therapy-refractory lethal prostate cancer and provide a druggable mechanism that may transcend into effective combined therapeutic strategies. SIGNIFICANCE: Our study shows that specialized eIF3B-dependent translation of specific mRNAs released upon downregulation of the master transcription factor MITF confers castration resistance and immune evasion in lethal prostate cancer. Pharmacologic targeting of this mechanism delays castration resistance and increases immune-checkpoint efficacy. This article is featured in Selected Articles from This Issue, p. 2489.

A Comparative Study of Intralesional Acyclovir vs Immunotherapy for Treatment of Viral Warts.

BACKGROUND: Viral warts are caused by human papillomavirus (HPV), are difficult to treat with conventional modalities, and are cosmetically disfiguring; hence, immunomodulators are being used. The viral origin of warts suggests the antiviral drug acyclovir as a potential therapeutic option. The current study compares the effect of intralesional acyclovir (nucleoside analogue) and intralesional purified protein derivative (PPD) (immunotherapy) in treating various viral warts. METHODOLOGY: Prospective observational comparative study was conducted to determine the efficacy of acyclovir, and PPD administered via the intralesional route in patients with viral warts. The study population was categorized into two groups. One group received intralesional acyclovir, and the other received intralesional PPD. Patients were followed-up with for three months. Outcomes considered in our study were recovery (complete, partial, and no recovery) and side effects like pain, burning sensation, and desquamation. Statistical analysis was carried out by coguide software. RESULTS: In our study total of 40 participants, 20 in each group were included. 25 and 15 were of age <30, and ≥ 30, respectively, while 20 were males, and 20 females. Our study reported 60%, and 30% of complete recovery with intralesional acyclovir treatment and intralesional PPD treatment, respectively, in the twelfth week. However, p-value > 0.05 represented no significance between groups. 90% in the acyclovir-treated group presented with pain, and 100% presented with burning sensation, while in the case of PPD-treated group, 60% presented no side effects and the rest 40% showed pain. CONCLUSIONS: Intralesional acyclovir is more effective in treating viral warts than PPD. The focus is to be laid on anticipated side effects.

CDK4: a master regulator of the cell cycle and its role in cancer.

The cell cycle is regulated in part by cyclins and their associated serine/threonine cyclin-dependent kinases, or CDKs. CDK4, in conjunction with the D-type cyclins, mediates progression through the G1 phase when the cell prepares to initiate DNA synthesis. Although Cdk4-null mutant mice are viable and cell proliferation is not significantly affected in vitro due to compensatory roles played by other CDKs, this gene plays a key role in mammalian development and cancer. This review discusses the role that CDK4 plays in cell cycle control, normal development and tumorigenesis as well as the current status and utility of approved small molecule CDK4/6 inhibitors that are currently being used as cancer therapeutics.

Immune non-interference and safety study of Vi-DT typhoid conjugate vaccine with a measles, mumps and rubella containing vaccine in 9-15 months old Nepalese infants.

BACKGROUND: Typhoid fever is a common disease in developing countries especially in the Indian subcontinent and Africa. The available typhoid conjugate vaccines (TCV) have been found to be highly immunogenic in infants and children less than 2 years of age. Many countries are planning to adopt TCV in their routine EPI programs around 9 months of age when measles containing vaccines are given. Therefore, Vi-DT TCV was tested in 9-15 months aged healthy infants in Nepal to demonstrate non-interference with a measles containing vaccine. METHODS: This was a randomized, open label, phase III study to assess the immune non-interference, safety, and reactogenicity of Vi-DT typhoid conjugate vaccine when given concomitantly with measles, mumps and rubella (MMR) vaccine. A total of 360 participants aged 9-15 months were enrolled and randomized equally into Vi-DT + MMR (180 participants) or MMR alone (180 participants) group and were evaluated for immunogenicity and safety 28 days post vaccination. RESULTS: Using the immunogenicity set, difference between proportions (95% CI) of the Vi-DT + MMR group vs MMR alone group were -2.73% (-8.85, 3.38), -3.19% (-11.25, 4.88) and 2.91% (-3.36, 9.18) for sero-positivity rate of anti-measles, anti-mumps and anti- rubella, respectively. Only the lower bound of the range in difference of the proportions for sero-positivity rate of anti-mumps did not satisfy the non-inferiority criteria as it was above the -10% limit, which may not be of clinical significance. These results were confirmed in the per protocol set. There were no safety concerns reported from the study and both Vi-DT + MMR and MMR alone groups were comparable in terms of solicited and unsolicited adverse events . CONCLUSIONS: Results indicated that there is non-interference of MMR vaccine with Vi-DT and Vi-DT conjugate vaccine could be considered as an addition to the EPI schedule among children at risk of contracting typhoid.

Dentinal translucency and width of cementum: predicting the age over 55 years in South Indian adults using extracted sectioned teeth.

The main intention of this article was to evaluate the reliability of root dentine translucency (RDT) and the width of cementum (CW) in indicating the age over 55 years in forensic and criminal investigations. 600 non-restored, single rooted teeth (300 males and 300 females) which were extracted for periodontal or orthodontic reasons were collected. Each tooth was sectioned longitudinally until the desired thickness (250 µm) was obtained. Both the length of the RDT and CW were measured from these unstained ground sections of teeth using ImageJ computer software. Pearson's correlation coefficient indicated a very strong and positive correlation for RDW and CW with age in both sexes. Cut-off values of RDT= 7.07 and CW= 52.06 were obtained using the maximum Youden's index. The value for the area under the curve (AUC) was 0.987 for RDT and 0.910 for CW was seen as indicating a very high discrimination. The performance of these cut-off values was tested in a separate sample of sectioned teeth (n= 300) and was analyzed using contingency tables in both sexes. The sensitivity was 88.2% and 92.3%, while specificity was 98.9% for RDT in males and females. For CW, the sensitivity was 96.1% and 90.3%, and specificity was 76.7% and 74.4% in males and females, respectively. Bayes post-test probability was 98.9% for RDT in both sexes, while 80.5% in males and 78% in females, for CW. Based on our study findings, it can be concluded that both variables have performed well in predicting the age over 55 years. Further research concerning the radiographic study of secondary dentine deposition to predict legal age thresholds would be a great benefit for living adults who require age estimation in civil proceedings.

TGFß reprograms TNF stimulation of macrophages towards a non-canonical pathway driving inflammatory osteoclastogenesis.

It is well-established that receptor activator of NF-κB ligand (RANKL) is the inducer of physiological osteoclast differentiation. However, the specific drivers and mechanisms driving inflammatory osteoclast differentiation under pathological conditions remain obscure. This is especially true given that inflammatory cytokines such as tumor necrosis factor (TNF) demonstrate little to no ability to directly drive osteoclast differentiation. Here, we found that transforming growth factor ß (TGFß) priming enables TNF to effectively induce osteoclastogenesis, independently of the canonical RANKL pathway. Lack of TGFß signaling in macrophages suppresses inflammatory, but not basal, osteoclastogenesis and bone resorption in vivo. Mechanistically, TGFß priming reprograms the macrophage response to TNF by remodeling chromatin accessibility and histone modifications, and enables TNF to induce a previously unrecognized non-canonical osteoclastogenic program, which includes suppression of the TNF-induced IRF1-IFNß-IFN-stimulated-gene axis, IRF8 degradation and B-Myb induction. These mechanisms are active in rheumatoid arthritis, in which TGFß level is elevated and correlates with osteoclast activity. Our findings identify a TGFß/TNF-driven inflammatory osteoclastogenic program, and may lead to development of selective treatments for inflammatory osteolysis.

Modified Conventional Root Canal Shaping Technique in Primary Teeth: An In Vivo Study.

Aim and objective: The purpose of the study was to compare the efficiency of modified conventional root canal shaping technique versus conventional shaping technique in primary teeth. Materials and methods: A total of 184 primary root canals (2 groups of 92 canals each) with two-thirds of root length were selected and randomly allocated into two different shaping techniques, i.e., Group I: conventional root canal shaping technique, Group II: modified root canal shaping technique. The quality of obturation (Coll and Sadrain, 1996) and presence or absences of voids were assessed by using radiographs. The recorded data was statistically analyzed. Results: Significant difference was seen between conventional and modified conventional techniques in underfilled and optimally filled canals. Conclusion: This modified cleaning and shaping technique can be considered as alternative to conventional instrumentation technique, as it improves quality of obturation and decreases the number of voids. How to cite this article: Reddy ER, Raju SS, Sandipamu T, et al. Modified Conventional Root Canal Shaping Technique In Primary Teeth: An In Vivo Study. Int J Clin Pediatr Dent 2022;15(S-1):S8-S11.

IHC expression of AKT in different grades of epithelial dysplasia: An in vitro study.

Introduction: Akt, also known as protein kinase B, is a serine/threonine-specific protein-kinase which plays a key role in multiple cellular processes such as glucose metabolism, apoptosis, transcription and cell migration. The activation of Akt is one of the most frequent alterations observed in human cancer and tumour cells. Akt regulates cellular survival and metabolism by binding and regulating many downstream effectors, e.g., Nuclear Factor-kB, murine double minute 2(MDM2). Aims: To evaluate and compare immunohistochemical expression of Akt in normal epithelium and different histological grades of epithelial dysplasias. Materials and Methodology: Forty paraffin-embedded tissue sections were used for the immunohistochemical study of which 10 cases of normal epithelium, 10 cases of each mild, moderate and severe epithelial dysplasia which were diagnosed by haematoxylin and eosin procedures. The tissue sections were immunohistochemically analysed for expression of Akt in different grades of epithelial dysplasia by using anti-Akt-1 monoclonal antibody. Statistical analysis was carried out using statistical package for social science (SPSS, V 10.5). The data were analysed using Chi-square test and P < 0.05 was considered statistically significant. Results and Conclusion: An overall significant difference was observed when normal tissues were compared with epithelial dysplasia with a Chi-square value of 21.04, but there was no statistical significance between the three grades of epithelial dysplasias. In conclusion, this study suggests that Akt-1 overexpression can be one of the useful diagnostic markers for predicting the potential behaviour of oral dysplasias transforming into oral squamous cell carcinoma (OSCC).

Maternal adropin levels in patients with gestational diabetes mellitus: a systematic review and meta-analysis.

BACKGROUND: Adropin is a regulatory protein with potential implications in energy homeostasis, glucose regulation, and insulin resistance. AIM: The aim of this meta-analysis was to compare the maternal serum/plasma adropin levels between gestational diabetes mellitus (GDM) patients and non-GDM controls. METHODS: Relevant studies were retrieved by online database and manual searching. The standardized mean differences (SMDs) with 95% confidence intervals (CIs) were obtained by a random-effects meta-analysis. A one-study leave-out sensitivity analysis and trimester-wise subgroup analysis were performed. RESULTS: A total of eight observations were included in this meta-analysis. The results based on random-effects meta-analysis indicated that adropin levels were significantly increased in GDM patients as compared to non-GDM controls (SMD = 2.41, 95% CI = 0.52-4.29, p= .01). The sensitivity analysis indicated that no single study had significantly influenced the overall outcome. CONCLUSIONS: The results indicate that maternal serum/plasma adropin concentrations were significantly higher in GDM patients as compared to non-GDM controls suggesting the potential associations of adropin in GDM. Despite this, further studies are needed to investigate the mechanistic, diagnostic and prognostic roles of trimester-wise adropin levels in GDM and associated fetal outcomes.

ETS-Related Gene Activation Preserves Adherens Junctions and Permeability in Microvascular Endothelial Cells.

ABSTRACT: ERG (ETS-related gene) is a member of the ETS (Erythroblast-transformation specific) family of transcription factors abundantly present in vascular endothelial cells. Recent studies demonstrate that ERG has important roles in blood vessel stability and angiogenesis. However, it is unclear how ERG is potentially involved in microvascular barrier functions and permeability. A wide variety of diseases and clinical conditions including trauma-hemorrhagic shock and burn injury are associated with microvascular dysfunctions, which causes excessive microvascular permeability, tissue edema and eventually, multiple organ dysfunction and death. The main purpose of this study was to determine the specific role of ERG in regulating microvascular permeability in human lung microvascular endothelial cells (HLMEC) and to evaluate if exogenous ERG will protect the barrier. The HLMECs were grown on Transwell inserts as monolayers and were transfected with ERG CRISPR/cas9 knockdown plasmid, ERG CRISPR activation plasmid, recombinant ERG protein or their respective controls. Recombinant vascular endothelial growth factor (VEGF) was used as an inducer of permeability for evaluating the effect of ERG activation on permeability. Changes in barrier integrity and permeability were studied using monolayer permeability assay and immunofluorescence of adherens junction proteins (VE-cadherin and ß-catenin) respectively. CRISPR/cas9-based ERG knockdown as well as VEGF treatment induced monolayer hyperpermeability, VE-cadherin, and ß-catenin junctional relocation and cytoskeletal F-actin stress fiber formation. CRISPR based ERG activation and recombinant ERG transfection attenuated VEGF-induced monolayer hyperpermeability. ERG activation preserved the adherens junctions and cytoskeleton. These results demonstrate that ERG is a potent regulator of barrier integrity and permeability in human lung microvascular endothelial cells and endogenously or exogenously enhancing ERG provides protection against barrier dysfunction and hyperpermeability.

Safety and immunogenicity of the Vi-DT typhoid conjugate vaccine in healthy volunteers in Nepal: an observer-blind, active-controlled, randomised, non-inferiority, phase 3 trial.

BACKGROUND: Typhoid fever is an endemic disease in many low-income and middle-income countries. The 2018 WHO position paper recommends that countries should consider typhoid vaccination in high-risk groups and for outbreak control. To address the typhoid vaccine supply and demand gap, a typhoid Vi polysaccharide-diphtheria toxoid (Vi-DT) conjugate vaccine development effort was undertaken to achieve WHO prequalification and contribute to the global supply of typhoid conjugate vaccine. The main aim of this study was to show immune non-inferiority of the Vi-DT vaccine compared with the WHO prequalified Vi polysaccharide-tetanus toxoid (Vi-TT) conjugate vaccine (Typbar TCV; Bharat Biotech India, Hyderabad, India) in participants of various ages from an endemic country. METHODS: We did an observer-blind, active-controlled, randomised, non-inferiority, phase 3 trial at four hospitals in Kathmandu, Dhulikhel, Dharan, and Nepalgunj in Nepal. Eligible participants were healthy individuals aged 6 months to 45 years for whom informed consent was obtained, were willing to follow the study procedures and were available for the duration of the study. Patients with an acute or chronic illness that could interfere with interpretation of the study endpoints, or who were involved in any other clinical trial were excluded. Participants were randomly assigned (1:1:1:1) by block randomisation (block size of four and eight), stratified by age (6 months to <2 years, 2 years to <18 years, and 18 years to 45 years), into one of four groups (A-D). Participants in groups A-C received a single dose (25 µg; 0·5 mL) of Vi-DT test vaccine via intramuscular injection from one of three good manufacturing practice lots (group A received lot 1, group B received lot 2, and group C received lot 3), and those in group D received a single dose (25 µg; 0·5 mL) of the Vi-TT vaccine via intramuscular injection. All participants, site staff (except for those who administered the study vaccines), and those assessing the outcomes were masked to group assignment. The co-primary endpoints were: (1) non-inferiority of immunogenicity of the Vi-DT vaccine (pooled groups A-C) versus the Vi-TT vaccine (group D), measured by the anti-Vi IgG seroconversion rate at 4 weeks after vaccination; and (2) the lot-to-lot consistency of the Vi-DT vaccine, measured by immune equivalence of the anti-Vi IgG geometric mean titre (GMT) at 4 weeks after receipt of the three Vi-DT vaccine lots (lot 1 vs lot 2, lot 1 vs lot 3, and lot 2 vs lot 3). Non-inferiority of the Vi-DT vaccine compared with the Vi-TT vaccine was shown if the lower limit of the 97·5% CI for the difference between the seroconversion rates in Vi-DT vaccine groups A-C combined versus Vi-TT vaccine group D was above the predefined non-inferiority margin of -10%. Lot-to-lot immune equivalence was shown if the upper and lower bounds of the two-sided 99·17% CI around the GMT ratio for each pairwise lot-to-lot comparison was between 0·67 and 1·50, which is the predefined equivalence margin recommended by WHO. The co-primary immunogenicity endpoints were assessed in all randomised participants who had received their assigned vaccine and had completed at least one post-baseline immunogenicity assessment. Safety was descriptively summarised by group and age strata, and was assessed in all participants who had received one dose of the investigational vaccine. The trial is registered with ClinicalTrials.gov, NCT03933098. FINDINGS: Between Nov 20, 2019, and March 10, 2020, 1854 individuals were screened, of whom 1800 were enrolled and randomly assigned to groups A-D (450 participants in each group). 1786 (99·2%; 443 in group A, 450 in group B, 447 in group C, and 446 in group D) were included in the immunogenicity assessments at 4 weeks post vaccination, and all 1800 participants were included in the safety analysis. In the immunogenicity analysis, the anti-Vi-IgG seroconversion rate in all age strata was 99·33% (97·5% CI 98·61 to 99·68; 1331 of 1340 participants) in Vi-DT vaccine groups A-C and 98·88% (97·10 to 99·57; 441 of 446) in Vi-TT vaccine group D. The difference in seroconversion rates between Vi-DT vaccine groups A-C combined versus Vi-TT group D was 0·47% (97·5% CI -0·68 to 1·61), indicating non-inferiority of the Vi-DT vaccine. Anti-Vi-IgG GMT ratios at 4 weeks post-vaccination were 1·02 (99·17% CI 0·85 to 1·22) for lot 1 versus lot 2, 1·02 (0·85 to 1·23) for lot 1 versus lot 3, and 1·01 (0·84 to 1·21) for lot 2 versus lot 3, indicating lot-to-lot equivalence according to the predefined, WHO-recommended equivalence margin. The proportion of participants reporting adverse events was similar between Vi-DT vaccine groups A-C and Vi-TT vaccine group D; 260 (19·3%) of 1350 participants in Vi-DT vaccine groups A-C and 115 (25·6%) of 450 in Vi-TT vaccine group D reported solicited adverse events within 7 days after vaccination, and 208 (15·4%) in Vi-DT vaccine groups A-C and 76 (16·9%) in Vi-TT vaccine group D reported unsolicited adverse events within 4 weeks after vaccination. Seven serious adverse events (four [0·3%] participants in Vi-DT vaccine groups A-C and three [0·7%] in Vi-TT vaccine group D), including one death in the Vi-TT vaccine group, were reported during the 24-week follow-up period, none of which were considered related to the investigational product. INTERPRETATION: When administered as a single dose, the Vi-DT test vaccine was safe, immunogenic, and non-inferior to the Vi-TT vaccine at 4 weeks post vaccination. Equivalent immunogenicity of the three lots of Vi-DT vaccine was also shown, supporting the manufacturing process of this vaccine. Once prequalified by WHO, this vaccine could be an option for purchase by UN agencies. FUNDING: The Bill & Melinda Gates Foundation. TRANSLATION: For the Nepali translation of the abstract see Supplementary Materials section.

Participatory cooking demonstrations: A distinctive learning approach towards positive health.

Background: Participatory cooking demonstration is a unique and effective way to teach nutritional concepts and basic cooking skills in a community setting. The present study attempted to develop the same in 4th-year nursing students through the intervention for a better nutritional health outcome. The objectives are to introduce and train nursing students on participatory cooking demonstrations in community households, evaluate the improvement of knowledge and self-efficacy of the participants, and assess the feedback of the study participants towards the intervention. Methodology: An educational intervention was carried out among BSc Nursing students in a tertiary healthcare institute from April to June 2019. A sample of 66 students were subjected to pre- and post-tests along with a self-efficacy evaluation and feedback survey. Results: Out of all, 91.1% were between 21 and 30 years, 77.8% belonged to rural areas, and 82% were in the lower-middle socioeconomic class. The knowledge was improved, and it was found to be statistically significant (P < 0.0001). The self-efficacy evaluation showed an enhancement of knowledge and awareness. The majority either strongly agreed or agreed that participatory cooking demonstrations helped them learn healthy cooking practices (80%), analyze specific nutritional problems (95.6%), and get hands-on experience in nutritional care (86.4%). The themes that emerged from qualitative data were discussed under liked, disliked aspects, challenges faced, and solutions offered. Conclusion: The hands-on sessions on participatory cooking demonstrations were successfully introduced and the knowledge and self-efficacy of the participants was improved. All participants were satisfied with the intervention as perceived by the participants.

Posiphen Reduces the Levels of Huntingtin Protein through Translation Suppression.

Posiphen tartrate (Posiphen) is an orally available small molecule that targets a conserved regulatory element in the mRNAs of amyloid precursor protein (APP) and α-synuclein (αSYN) and inhibits their translation. APP and αSYN can cause neurodegeneration when their aggregates induce neurotoxicity. Therefore, Posiphen is a promising drug candidate for neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease. Posiphen's safety has been demonstrated in three independent phase I clinical trials. Moreover, in a proof of concept study, Posiphen lowered neurotoxic proteins and inflammatory markers in cerebrospinal fluid of mild cognitive impaired patients. Herein we investigated whether Posiphen reduced the expression of other proteins, as assessed by stable isotope labeling with amino acids in cell culture (SILAC) followed by mass spectrometry (MS)-based proteomics. Neuroblastoma SH-SY5Y cells, an in vitro model of neuronal function, were used for the SILAC protein profiling response. Proteins whose expression was altered by Posiphen treatment were characterized for biological functions, pathways and networks analysis. The most significantly affected pathway was the Huntington's disease signaling pathway, which, along with huntingtin (HTT) protein, was down-regulated by Posiphen in the SH-SY5Y cells. The downregulation of HTT protein by Posiphen was confirmed by quantitative Western blotting and immunofluorescence. Unchanged mRNA levels of HTT and a comparable decay rate of HTT proteins after Posiphen treatment supported the coclusion that Posiphen reduced HTT via downregulation of the translation of HTT mRNA. Meanwhile, the downregulation of APP and αSYN proteins by Posiphen was also confirmed. The mRNAs encoding HTT, APP and αSYN contain an atypical iron response element (IRE) in their 5'-untranslated regions (5'-UTRs) that bind iron regulatory protein 1 (IRP1), and Posiphen specifically bound this complex. Conversely, Posiphen did not bind the IRP1/IRE complex of mRNAs with canonical IREs, and the translation of these mRNAs was not affected by Posiphen. Taken together, Posiphen shows high affinity binding to the IRE/IRP1 complex of mRNAs with an atypical IRE stem loop, inducing their translation suppression, including the mRNAs of neurotoxic proteins APP, αSYN and HTT.

Simultaneous CK2/TNIK/DYRK1 inhibition by 108600 suppresses triple negative breast cancer stem cells and chemotherapy-resistant disease.

Triple negative breast cancer (TNBC) remains challenging because of heterogeneous responses to chemotherapy. Incomplete response is associated with a greater risk of metastatic progression. Therefore, treatments that target chemotherapy-resistant TNBC and enhance chemosensitivity would improve outcomes for these high-risk patients. Breast cancer stem cell-like cells (BCSCs) have been proposed to represent a chemotherapy-resistant subpopulation responsible for tumor initiation, progression and metastases. Targeting this population could lead to improved TNBC disease control. Here, we describe a novel multi-kinase inhibitor, 108600, that targets the TNBC BCSC population. 108600 treatment suppresses growth, colony and mammosphere forming capacity of BCSCs and induces G2M arrest and apoptosis of TNBC cells. In vivo, 108600 treatment of mice bearing triple negative tumors results in the induction of apoptosis and overcomes chemotherapy resistance. Finally, treatment with 108600 and chemotherapy suppresses growth of pre-established TNBC metastases, providing additional support for the clinical translation of this agent to clinical trials.

Evaluation of stress in three different fiber posts with two-dimensional finite element analysis.

INTRODUCTION: The primary function of corono-radicular post is to provide retention for the core and to reinforce and to replace the remaining coronal tooth structure. There is considerable controversy regarding optimal choice of the material. An ideal post system should exhibit fracture resistance higher than the average masticatory forces. Finite elemental analysis (FEA) method facilitates precise analysis of the distribution and magnitude of stresses at any point of complex and irregular structures. Thus, this FEA study has been undertaken to evaluate the fracture stress distribution patterns in three fiber posts, viz., carbon, glass fiber, and everStick with an FEA. MATERIALS AND METHODS: The FE stress analysis was performed with the FE software program (CATIA). Three two-dimensional FEA models of central incisor were simulated, and elastic moduli and Poisson's ratio of all the materials were fed to the software. For all the models, a 200 N vertical force was applied on the lingual surface of the tooth at an angle of 45°. Stress concentration and distribution were evaluated and noted down for all the models. To evaluate the stresses within the restored tooth, the modified von Mises failure criterion was used. The equivalent stresses found in the tested models were compared with the tensile strength of the respective materials. Contact stresses in the luting cement-dentin interface were calculated. RESULTS: Finite element method revealed that maximum stress concentration was at the point of stress application. The stress value was highest in carbon fiber post followed by glass fiber post and least stresses found in everStick post. Maximum stress was observed at the labial surfaces of crown. However, the stress values and distribution were more homogenous in everStick post. CONCLUSION: The present findings suggest that everStick post has uniform stress distribution within tooth structure.

Pyrimidine-2,4-dione targets STAT3 signaling pathway to induce cytotoxicity in hepatocellular carcinoma cells.

Signal transducer and activator of transcription 3 (STAT3) is a tumorigenic transcription factor that is persistently activated in various human cancers including hepatocellular carcinoma (HCC). Therefore, STAT3 is considered as a prominent target to counteract the uncontrolled proliferation of cancer cells. In the present report, pyrimidine-2,4-diones (N-methyluracil derivatives) (MNK1-MNK14) were synthesized in an ionic liquid (BMIm PF6) medium employing a ligand-free Suzuki-Miyaura cross-coupling process. Among the 14 derivatives, compound MNK8 showed good cytotoxicity towards both the tested cell lines and did not display a toxic effect against normal hepatocytes (LO2). MNK8 significantly increased the Sub-G1 cell count in both cell lines and the cytotoxic effect of MNK8 was found to be mediated through the suppression of constitutive phosphorylation of STAT3Y705. It also decreased the DNA interaction ability of nuclear STAT3 in HCC cells. MNK8 downregulated the levels of apoptosis-related proteins (such as Bcl-2, cyclin D1, survivin) and increased cleaved caspase-3 inferring the apoptogenic effect of MNK8. It also reduced the CXCL12-triggered cell migration and invasion in in vitro assay systems. Overall, MNK8 has been demonstrated as a new inhibitor of STAT3 signaling cascade in HCC cells.

Adropin levels and its associations as a fat-burning hormone in patients with polycystic ovary syndrome: a correlational meta-analysis.

AIMS: Adropin is a peptide hormone with potential implications in patients with polycystic ovary syndrome (PCOS). The aim of this meta-analysis was to compare the circulating (serum/plasma) and follicular fluid adropin levels between human PCOS patients and non-PCOS controls. METHODS: Relevant studies were retrieved by online database and manual searching. The standardized mean differences (SMDs) with 95% confidence intervals (CIs) were obtained by a random-effects meta-analysis. Meta-analysis of correlations was performed for the associations of adropin with anthropometric, lipid, insulin resistance and hormonal covariates. The funnel plot analysis with Begg's and Egger's tests was used for publication bias. RESULTS: A total of 9 studies were included in this meta-analysis. The results indicated that the adropin levels were significantly decreased in PCOS patients as compared to non-PCOS controls (SMD = -1.87, 95% CI = -2.55 to -1.18, p < .0001). This decrease was more evident in overweight PCOS patients than their normoweight counterparts (SMD = -0.55, 95% CI = -0.80 to -0.30, p < .0001). A one-study leave-out sensitivity analysis indicated that no single study had a significant influence on the overall outcome, suggesting the robustness of this meta-analysis. There were significant associations of decreased adropin levels with the body mass index, dyslipidemia and insulin resistance in PCOS. CONCLUSION: Adropin levels are significantly reduced in PCOS patients compared to controls, and this decrease was more evident in overweight PCOS patients.

Immunohistochemical evaluation of myofibroblasts in odontogenic keratocyst, dentigerous cyst and different clinical variants of ameloblastoma: A comparative study.

BACKGROUND: Odontogenic cysts and tumors exhibit varying degrees of aggressiveness in their biological behavior. Odontogenic keratocyst (OKC), dentigerous cyst (DC), ameloblastoma are most common odontogenic cysts and tumors to occur in the oral cavity. Myofibroblasts (MFs) in the connective tissue stroma participate in the matrix degradation process by secreting matrix metalloproteinase 2, transforming growth factor beta1 and may contribute to variation in their biological behavior. Its activity is identified by alpha-smooth muscle actin (α-SMA) marker. With this background, the present study aims to evaluate the frequency of MFs using α-SMA to determine the biological behavior of OKC, DC, and different clinical variants of ameloblastoma. MATERIALS AND METHODS: A retrospective study was carried out with total of 60 samples which include 10 cases each of OKC, DC, 30 cases of different clinical variants of ameloblastomas and 10 normal mucosa taken as controls. All are stained immunohistochemically using α-SMA and were analyzed for the same. Comparison between more than 2 groups done by one way analysis of variance test with the level of significance of P ≤ 0.0001, i.e., <0.05. RESULTS: Statistically significant difference in the mean number of MFs observed between certain groups, with higher mean number in solid ameloblastoma (SA) (32.45) followed by OKC (28.79), unicystic ameloblastoma (24.53), desmoplastic ameloblastoma (7.44), and DC (1.72). CONCLUSION: Higher frequency of MFs noticed in SA, OKC which are key cells for connective tissue remodeling by interacting with epithelial cells and other connective tissue cells to facilitate progression of cysts and tumors thereby contributing to their biological behavior.

Novel induction of CD40 expression by tumor cells with RAS/RAF/PI3K pathway inhibition augments response to checkpoint blockade.

BACKGROUND: While immune checkpoint blockade (ICB) is the current first-line treatment for metastatic melanoma, it is effective for ~ 52% of patients and has dangerous side effects. The objective here was to identify the feasibility and mechanism of RAS/RAF/PI3K pathway inhibition in melanoma to sensitize tumors to ICB therapy. METHODS: Rigosertib (RGS) is a non-ATP-competitive small molecule RAS mimetic. RGS monotherapy or in combination therapy with ICB were investigated using immunocompetent mouse models of BRAFwt and BRAFmut melanoma and analyzed in reference to patient data. RESULTS: RGS treatment (300 mg/kg) was well tolerated in mice and resulted in ~ 50% inhibition of tumor growth as monotherapy and ~ 70% inhibition in combination with αPD1 + αCTLA4. RGS-induced tumor growth inhibition depends on CD40 upregulation in melanoma cells followed by immunogenic cell death, leading to enriched dendritic cells and activated T cells in the tumor microenvironment. The RGS-initiated tumor suppression was partially reversed by either knockdown of CD40 expression in melanoma cells or depletion of CD8+ cytotoxic T cells. Treatment with either dabrafenib and trametinib or with RGS, increased CD40+SOX10+ melanoma cells in the tumors of melanoma patients and patient-derived xenografts. High CD40 expression level correlates with beneficial T-cell responses and better survival in a TCGA dataset from melanoma patients. Expression of CD40 by melanoma cells is associated with therapeutic response to RAF/MEK inhibition and ICB. CONCLUSIONS: Our data support the therapeutic use of RGS + αPD1 + αCTLA4 in RAS/RAF/PI3K pathway-activated melanomas and point to the need for clinical trials of RGS + ICB for melanoma patients who do not respond to ICB alone. TRIAL REGISTRATION: NCT01205815 (Sept 17, 2010).